Personal Care Products and Domestic Cleaning Products

Final Report to DWI September 2020

Personal Care Products and

Domestic Cleaning Products –

Toxicological Assessment of

Prioritised List of Chemicals (Ref:

DWI 70/2/331)

FINAL REPORT

Report to Defra/DWI IEH Consulting Ltd.|September 2020


PCPs and DCPs – toxicological assessment of

prioritised list of chemicals

A Report to DWI by IEH Consulting Ltd.

Prepared by: Ruth Bevan, Sarah Bull

Reviewed by: Camilla Alexander-White, Paul Rumsby

Edited by: Paul Harrison

September 2020

Disclaimer The views expressed in this report are those of the authors alone, representing their

expert opinion based on the review of the available information and their

understanding drawn from publications identified from targeted literature searches.


Contents

Contents Abbreviations ............................................................................................................................. 8

Executive Summary .................................................................................................................... 9

1.0 Introduction ....................................................................................................................... 11

1.1 Project Background ............................................................................................................ 11

1.2 Personal Care Products ...................................................................................................... 11

1.3 Domestic Cleaning Products .............................................................................................. 11

1.4 Selection of PCPs and DCPs of concern ............................................................................. 12

1.5 Aims and objectives ........................................................................................................... 13

2.0 Methodology ...................................................................................................................... 13

2.1 Literature search ................................................................................................................ 13

2.2 Screening of literature ....................................................................................................... 13

2.3 Risk assessment process .................................................................................................... 14

2.3.1 Hazard identification ............................................................................................... 14

2.3.2 Dose-response assessment ..................................................................................... 14

2.3.3 Exposure assessment .............................................................................................. 15

2.3.4 Risk characterisation ............................................................................................... 15

General references .................................................................................................................. 16

3.0 Results ................................................................................................................................ 18

3.1 Identification of data ......................................................................................................... 18

3.2 Human health risk assessments for exposure via drinking water ..................................... 19

Section A: 1,2,3-benzotriazole ................................................................................................. 20

Section A1.0 Chemical identification, use and potential human exposure ............................. 20

Section A1.1 Reasons for consideration .......................................................................... 20

Section A1.2 Identification and physicochemical properties. ......................................... 20

Section A1.3 Hazard classifications .................................................................................. 20

Section A1.4 Occurrence, production and use. ............................................................... 20

Section A1.5 Human exposure ......................................................................................... 21

Section A2.0 Human relevant health effects ........................................................................... 21

Section A2.1 Toxicokinetics.............................................................................................. 21

Section A2.2 Acute toxicity .............................................................................................. 22

Section A2.3 Repeat dose toxicity ................................................................................... 22


Section A3.0 Identification of relevant point(s) of departure for use in risk assessment ....... 27

Section A3.1 Critical endpoint for human health risk assessment purposes .................. 27

Section A3.2 Current guideline values ............................................................................. 30

Section A3.3 Identification of POD to be used for the risk assessment .......................... 30

Section A4.0 – Drinking water risk assessment ....................................................................... 30

References ............................................................................................................................... 31

Section B: 1H-benzotriazole, 4(or 5)-methyl- .......................................................................... 33

Section B1.0 Chemical identification, use and potential human exposure ............................. 33

Section B1.1 Reasons for consideration .......................................................................... 33

Section B1.2 Identification and physicochemical properties. ......................................... 33

Section B1.3 Hazard classifications .................................................................................. 33

Section B1.4 Occurrence, production and use. ............................................................... 33

Section B1.5 Human exposure ......................................................................................... 34

Section B2.0 Human relevant health effects ........................................................................... 35

Section B2.1 Toxicokinetics .............................................................................................. 35

Section B2.2 Acute toxicity .............................................................................................. 35

Section B2.3 Repeat dose toxicity.................................................................................... 35

Section B3.0 Identification of relevant point(s) of departure for use in risk assessment ....... 38

Section B3.1 Critical endpoint for human health risk assessment purposes .................. 38

Section B3.2 Current health-based guidance values ....................................................... 41

Section B3.3 Identification of POD to be used for the risk assessment. ......................... 41

Section B4.0 Drinking water risk assessment .......................................................................... 41

References ............................................................................................................................... 42

Section C: (1-hydroxyethylidene) diphosphonic acid (HEDP) .................................................. 43

Section C1.0 Chemical identification, use and potential human exposure ............................. 43

Section C1.1 Reasons for consideration .......................................................................... 43

Section C1.2 Identification and physicochemical properties. ......................................... 43

Section C1.3 Hazard classifications .................................................................................. 44

Section C1.4 Occurrence, production and use. ............................................................... 44

Section C1.5 Human exposure ......................................................................................... 44

Section C2.0 Human relevant health effects ........................................................................... 44

Section C2.1 Toxicokinetics .............................................................................................. 44

Section C2.2 Acute toxicity .............................................................................................. 45

Section C2.3 Repeat dose toxicity .................................................................................... 45


Section C3.0 Identification of relevant point(s) of departure for use in risk assessment ....... 52

Section C3.1 Critical endpoints for human health risk assessment purposes ................. 52

Section C3.2 Current health-based guidance values ....................................................... 55

Section C3.3 Identification of POD to be used for the risk assessment .......................... 55

Section C4.0 Drinking water risk assessment .......................................................................... 55

References ............................................................................................................................... 56

Section D: Diethylenetriamine penta(methylene phosphonic acid) (DTPMP) ........................ 57

Section D1.0 Chemical identification, use and potential human exposure ............................ 57

Section D1.1 Reasons for consideration .......................................................................... 57

Section D1.2 Identification and physicochemical properties .......................................... 57

Section D1.3 Hazard classifications.................................................................................. 58

Section D1.4 Occurrence, production and use ................................................................ 58

Section D1.5 Human exposure ........................................................................................ 59

Section D2.0 Human relevant health effects ........................................................................... 59

Section D2.1 Toxicokinetics ............................................................................................. 59

Section D2.2 Acute toxicity .............................................................................................. 59

Section D2.3 Repeat dose toxicity ................................................................................... 60

Section D3.0 Identification of relevant point(s) of departure for use in risk assessment ....... 64

Section D3.1 Critical endpoints for human health risk assessment purposes ........................ 64

Section D4.0 Drinking water risk assessment .......................................................................... 67

References ............................................................................................................................... 68

Section E: Aminotris(methylene phosphonic acid) (ATMP) ..................................................... 70

Section E1.0 Chemical identification, use and potential human exposure ............................. 70

Section E1.1 Reasons for consideration........................................................................... 70

Section E1.2 Identification and physicochemical properties. .......................................... 70

Section E1.3 Hazard classifications .................................................................................. 71

Section E1.4 Occurrence, production and use................................................................. 71

Section E1.5 Human exposure ......................................................................................... 71

Section E2.0 Human relevant health effects ........................................................................... 71

Section E2.1 Toxicokinetics .............................................................................................. 71

Section E2.2 Acute toxicity .............................................................................................. 71

Section E2.3 Repeat dose toxicity .................................................................................... 72

Section E3.0 Identification of relevant point(s) of departure for use in risk assessment ....... 77

Section E3.1 Critical endpoints for human health risk assessment purposes ................. 77


Section E3.2 Current health-based guidance values ....................................................... 80

Section E3.3 Identification of POD to be used for the risk assessment. ......................... 80

Section E4.0 Drinking water risk assessment .......................................................................... 80

References ............................................................................................................................... 81

Section F: 2-(2-butoxyethoxy)ethanol (DEGBE) ....................................................................... 82

Section F1.0 Chemical identification, use and potential human exposure ............................. 82

Section F1.1 Reasons for consideration ........................................................................... 82

Section F1.2 Identification and physicochemical properties. .......................................... 82

Section F1.3 Hazard classifications .................................................................................. 82

Section F1.4 Occurrence, production and use ................................................................. 82

Section F1.5 Human exposure ......................................................................................... 83

Section F2.0 Human relevant health effects ........................................................................... 83

Section F2.1 Toxicokinetics .............................................................................................. 83

Section F2.2 Acute toxicity ............................................................................................... 84

Section F2.3 Repeat dose toxicity .................................................................................... 84

Section F2.3.2 Irritancy and corrosivity ........................................................................... 85

Section F3.0 Identification of relevant point(s) of departure for use in risk assessment ....... 88

Section F3.1 Critical endpoints for human health risk assessment purposes ................. 88

Section F3.2 Current health-based guidance values ....................................................... 92

Section F3.3 Identification of POD to be used for the risk assessment........................... 92

Section F4.0 Drinking water risk assessment........................................................................... 92

References ............................................................................................................................... 94

Section G: Linear alkylbenzene sulphonate (LAS) .................................................................... 96

Section G1.0 Chemical identification, use and potential human exposure ............................ 96

Section G1.1 Reasons for consideration .......................................................................... 96

Section G1.2 Identification and physicochemical properties .......................................... 96

Sources-WRc 2014; PubChem; ChemID Plus; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020 ....................................... 97

Section G1.3 Hazard classifications ................................................................................. 97

Section G1.4 Occurrence, production and use ................................................................ 97

Section G1.5 Human exposure ........................................................................................ 97

Section G2.0 Human relevant health effects ........................................................................... 97

Section G2.1 Toxicokinetics ............................................................................................. 97

Section G2.2 Acute toxicity .............................................................................................. 98


Section G2.3 Repeat dose toxicity ................................................................................... 99

Section G3.0 Identification of relevant point(s) of departure for use in risk assessment..... 107

Section G3.1 Critical endpoints for human health risk assessment purposes .............. 107

Section G3.2 Current health-based guidance values ..................................................... 114

Section G 3.3 Identification of POD to be used for the risk assessment ....................... 114

Section G4.0 Drinking water risk assessment ........................................................................ 114

References ............................................................................................................................. 116

Section H: Cocamidopropyl betaine (CAPB) .......................................................................... 117

Section H1.0 Chemical identification, use and potential human exposure .......................... 117

Section H1.1 Reasons for consideration ........................................................................ 117

Section H1.2 Identification and physicochemical properties ........................................ 117

Section H1.3 Hazard classifications ............................................................................... 118

Section H1.4 Occurrence, production and use. ............................................................. 119

Section H1.5 Human exposure ...................................................................................... 119

Section H2.0 Human relevant health effects ......................................................................... 119

Section H2.1 Toxicokinetics ........................................................................................... 119

Section H2.2 Acute toxicity ............................................................................................ 120

Section H2.3 Repeat dose toxicity ................................................................................. 120

Section H3.0 Identification of relevant point(s) of departure for use in risk assessment..... 124

Section H3.1 Critical endpoint for human health risk assessment purposes ................ 124

Section H3.2 Current health-based guidance values ..................................................... 127

Section H3.3 Identification of POD to be used for the risk assessment ........................ 127

Section H4.0 Drinking water risk assessment ........................................................................ 127

References ............................................................................................................................. 128

4.0 Risk assessment outcomes and conclusions .................................................................... 131

Annex A – Literature search strategy .................................................................................... 135

Annex B – Secondary screening ............................................................................................. 138


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Abbreviations

ADI acceptable daily intake AF assessment factor (also known as uncertainty factor (UF)) ATMP aminotris(methylene phosphonic acid) BMD benchmark dose BMDL benchmark dose level (95 % lower confidence limit of the BMD) BMR benchmark response CAPB cocamidopropyl betaine DCP domestic cleaning product DEGBE 2-(2-butoxyethoxy)ethanol) DSL domestic substances list DTPMP diethylenetriamine penta(methylene phosphonic acid ECHA European chemicals agency GLP good laboratory practice HPRT hypoxanthine-guanine phosphoribosyl transferase HBGV health-based guidance value HEDP 1-hydroxy-ethylidene 1, 1-diphosphonic acid HQ hazard quotient i.p. intraperitoneal i.v. intravenous LAS linear alkylbenzene sulphonate LD50 lethal dose (50 % of population) LOAEL lowest observed adverse effects level MOE margin of exposure NOAEL no observed adverse effect level OECD organisation for economic co-operation and development PCP personal care product POD point of departure QSAR models quantitative structure–activity relationship models S9 fraction product of an organ tissue homogenate (usually liver) used

in biological assays to stimulate metabolism TDI tolerable daily intake TWI tolerable weekly intake

https://en.wikipedia.org/wiki/Homogenization_(biology)

https://en.wikipedia.org/wiki/Bioassay


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Executive Summary

Personal care products (PCPs) and Domestic Cleaning products (DCPs) contain a wide range of chemicals, according to their intended purpose. Patterns of use between different PCPs and/or DCPs also differ and, as a consequence, the duration and levels of human exposure to the chemicals present can vary significantly. PCPs are categorised by their use and include ‘leave-on’ products such as cosmetics, moisturisers, body sprays and deodorants’, ‘rinse-off’ products including shampoos, soaps, shower gels and shaving gels, and ‘oral care’ products such as toothpaste and mouthwashes. DCPs are classed as those used for ‘laundry/dish care’ including dishwasher tablets/powders, washing up liquids and laundry powders, ‘surface cleaning’ such as kitchen and bathroom spray cleaners, ‘air care’ including air fresheners and fragrances, and ‘floor care’ such as hard surface cleaners and carpet shampoo. Importantly, the majority of PCPs/DCPs used in the home are disposed of down the drain, thus entering the sewerage system with the potential to reach drinking water supplies.

A study commissioned by the Drinking Water Inspectorate (DWI) and conducted in 2014 investigated the potential of 690 chemicals used in PCPs and DCPs to be present in drinking water (WRc, 2014). From this, 33 chemicals were identified as having an increased potential for reaching drinking water and, for 10 of these, exposure from drinking water and bathing was considered higher than would be expected from normal use of the products. The study reported here expands on these previous findings by carrying out a risk assessment to determine if the toxicological properties of the chemicals are of concern with respect to exposure of humans via drinking water. EDTA and boric acid were eliminated from the priority list as guideline values indicated no safety concern at the levels identified. The eight chemicals included in the risk assessment process were: 1,2,3-benzotriazole; 1H-benzotriazole, 4(or 5)-methyl-; (1-hydroxyethylidene) diphosphonic acid (HEDP); diethylenetriamine penta (methylene phosphonic acid) (DTPMP); amino tris(methylene phosphonic acid) (ATMP); 2-(2-butoxyethoxy) ethanol (DEGBE); linear alkylbenzene sulphonate (LAS); cocamidopropyl betaine (CAPB).

Risk assessment was carried out by comparing potential intake (exposure) values from drinking water with toxicologically-derived health-based guidance values (HBGV). Hazard identification and characterisation was achieved for each chemical through a critical assessment of publicly available human and experimental literature. This allowed identification of a point of departure (POD) to which appropriate (chemical specific) assessment factors (AF) were applied to derive a tolerable daily intake (TDI). Exposure levels were determined using default body weight and consumption parameters for adults, children and infants, as advised by WHO (2017), with measured, where available, or modelled concentrations in drinking water, determined in the previous study (WRc, 2014). Risk was characterised through calculation of the hazard quotient (HQ), where intake was compared against the TDI, and calculation of the drinking water equivalent level (DWEL). Margin of exposure (MOE) was also calculated, for risk communication purposes, to illustrate the degree of ‘safety’ between estimated intake levels and the levels associated with adverse effects.

Hazard identification and characterisation was possible for all chemicals, however the availability of chronic toxicity studies was generally limited. All TDIs, with the exception of that for DEGBE, were derived from NOAELs and in several cases (HEDP, ATMP, 1,2,3-


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benzotriazole) this was the maximum dose tested in the study. Where available, the exposure assessment utilised measured data from the previous study (WRc, 2014); however, when unavailable, the maximum modelled levels in drinking water were used as a ‘worst-case’ scenario for risk assessment purposes. Estimated intakes of each chemical from drinking water were calculated for receptors in three age groups, to allow for the effect of body weight on intake levels, with infants being considered as potentially the most sensitive age group for risk assessment purposes. For all age groups intake was below the calculated TDI and all HQ’s were <1. The MOEs were also all >100, indicating that it was unlikely that any of the chemicals would be of concern following exposure via drinking water. Due to a lack of data concerning relative exposure from other sources, an assumption was made that total exposure to a chemical was from drinking water. In reality, other sources of exposure may occur through intended product use; however, the MOEs are sufficiently large to not be eroded to any significant extent if additional exposures were also taken into account.

Taken together, the evidence presented in this risk assessment indicates that for the chemicals of interest the levels that are potentially present in drinking water due to normal use of PCPs and DCPS are not anticipated to pose an appreciable risk to public health.


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1.0 Introduction

1.1 Project Background

Personal care products (PCPs) and Domestic Cleaning products (DCPs) contain a wide range of chemicals, depending on their intended purpose. Patterns of use between different PCPs and/or DCPs also differ and, as a consequence, the duration and levels of human exposure to the chemicals present can vary significantly. PCPs1 are categorised by their use and include ‘leave-on’ products such as cosmetics, moisturisers, body sprays and deodorants’, ‘rinse-off’ products including shampoos, soaps, shower gels and shaving gels, and ‘oral care’ products such as toothpaste and mouthwashes. DCPs2 are classed as those used for ‘laundry/dish care’ including dishwasher tablets/powders, washing up liquids and laundry powders, ‘surface cleaning’ such as kitchen and bathroom spray cleaners, ‘air care’ including air fresheners and fragrances, and ‘floor care’ such as hard surface cleaners and carpet shampoo. Importantly, the majority of PCPs/DCPs used in the home are disposed of down the drain, thus entering the sewerage system with the potential to reach drinking water supplies.

1.2 Personal Care Products

PCPs include ‘leave-on’ products such as cosmetics, moisturisers, some hair conditioners, fragrances, body sprays and deodorants. These are considered to have multiple routes into the sewerage system, both directly during application and removal from the human body and indirectly following transfer to hard surfaces (sinks and basins) or to clothing (washed off during laundering). ‘Rinse-off’ products include shampoos and conditioners, soaps, bath and shower gels and shaving gels, where application is for a limited time and and normal use leads to direct inputs to the sewerage system. It is also common for consumers to rinse containers, once empty, prior to recycling. A further type of PCP are ‘oral care’ products such as toothpaste and mouthwashes. These are again designed to have a limited contact time and are disposed of directly to the sewerage system. Hair dyes contain both ‘leave-on’ (colour) and ‘rinse-off’ (chemicals used to deliver the dye) elements, with both entering the sewerage system at different time points after use.

1.3 Domestic Cleaning Products

DCPs are classed as those used for laundry/dish care (for example, dishwasher tablets and powders, washing up liquids and laundry powders, liquids and softeners or tablets), surface cleaning (for example, kitchen and bathroom spray cleaners gel and abrasive cleaners, furniture polishes and waxes), air care (for example, air fresheners and fragrances) or floor care (for example, hard surface cleaners and carpet shampoo). Most laundry/dish and floor care products are ‘rinse-off’ products that are diluted with water, and the majority are discharged directly into the sewerage system after use. Some surface cleaners are designed to leave a residue (for example waxes), but most will be washed into the sewerage system

1 https://www.lawinsider.com/dictionary/personal-care-product 2 https://www.lawinsider.com/dictionary/household-cleansing-product


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directly on or following use. Air care products are most likely to be deposited onto hard surfaces or carpets, with limited entry to the sewerage system.

1.4 Selection of PCPs and DCPs of concern

A study conducted in 2014 investigated the potential of 690 chemicals used in PCPs and DCPs to be present in drinking water (WRc, 2014). Of the total number, 33 chemicals were identified as having an increased potential for reaching drinking water. For these, an exposure assessment was carried out to determine if drinking water consumers could be exposed to higher concentrations from drinking water and bathing than would be expected from normal use of the products. Some consumers of drinking water may have made a deliberate choice not to use PCPs and DCPs, however they may unknowingly still be exposed to traces of the chemicals present via consumption of drinking water. The exposure assessment identified 10 chemicals where this might apply (WRc, 2014).

This project aims to expand on these previous findings by carrying out a risk assessment to determine if the toxicological properties of 8 of these 10 chemicals are of concern following the exposure of humans via drinking water (WRc, 2014). Two of the chemicals, EDTA (CAS No. 6381-92-6) and boric acid (CAS No. 10043-35-3) have guideline values that indicate no safety concerns at the levels reported in the 2014 study and therefore are not included for further risk assessment here. Table 1 details the 8 chemicals (in priority order3) for which a risk assessment is carried out in this report.

Table 1: Prioritised* list of chemicals for risk assessment

Chemical CAS No. Category Product types

1 1,2,3-Benzotriazole 95-14-7 Corrosion inhibitor Dishwashing

2 1H-Benzotriazole, 4(or 5)-methyl- 29385-43-1 Corrosion inhibitor Dishwashing

3 (1-hydroxyethylidene) diphosphonic acid (HEDP)

2809-21-4 Phosphonate Dish, laundry and household cleaning

4 Diethylenetriamine penta (methylene phosphonic acid) (DTPMP)


5 Amino tris(methylene phosphonic acid) (ATMP)


6 2-(2-butoxyethoxy) ethanol (DEGBE) 112-34-5 Solvent Household cleaners

7 Linear alkylbenzene sulphonate (LAS) 68411-30-3 Anionic surfactant Detergents

8 Cocamidopropyl betaine (CAPB)

61789-40-0, 83138-08-3, 86438-79-1

Amphoteric surfactant

Foam booster in shampoo, hand soap and cosmetics.

*as reported by WRc (2014).

3 Prioritisation was based on the following parameters: Known to have/may have adverse effects in humans; Not removed by wastewater treatment processes; Not easily removed by drinking water treatment processes (it is acknowledged that some drinking water has limited treatment, such as groundwater); level of abiotic degradation (hydrolysis and photolysis); Level of abiotic degradation (hydrolysis and photolysis); Level of occurrence in raw water; Level of occurrence in drinking water; Usage statistics.


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1.5 Aims and objectives

This report describes the outcome of risk assessment of 8 prioritised chemicals in PCPs and DCPs following exposure of humans via drinking water. This was achieved firstly by undertaking a toxicological evaluation for each to identify a point(s) of departure (PODs) that could be used to determine a health-based guidance value (HBGV). In the second stage, calculated intakes for each chemical were compared to the HBGV to assess the human health risk (ECHA, 2016).

2.0 Methodology

2.1 Literature search

Information was sourced from a number of scientific and bibliographic databases in addition to publicly available sites and grey literature (i.e. reports from government, authoritative bodies and industry). Search terms were derived for each of the 8 prioritised chemicals. Adaptations were made to the search terms to ensure that the most appropriate data were identified. Exclusion criteria were developed for each chemical to ensure only relevant publications were captured (Annex A, Table A1). Searches were not restricted by date and were carried out between January and March 2020.

Of primary consideration were the toxicological data, derived from the following toxicology databases: hazardous substances databank (HSDB); International Uniform Chemical Information Database (IUCLID); Toxicology Data Network (TOXNET); ToxPlanet; Cosmetic Organic Standard (COSMOS); Human and Environmental Risk Assessment (HERA); and The European Chemicals Agency (ECHA; REACH). In addition, opinions from authoritative bodies such as the US Food and Drug Administration (FDA), Agency for Toxic Substances and Disease Registry (ATSDR), US Environmental Protection Agency (EPA) and The European Food Safety Authority (EFSA) were also interrogated. Primary literature was identified using PubMed / Science Direct, where data from other sources was scarce or unavailable. A focus was given to toxicity endpoints relating to chronic low-level exposure, as this better represented the exposure scenario from drinking water; however, acute data was noted when appropriate. Searches included: target organ toxicity, mutagenicity, carcinogenicity, reproductive and developmental toxicity, immunotoxicity, neurotoxicity and endocrine disrupting activity in animal and human studies and included in vitro studies. Any published PODs and HBGVs for the chemicals of interest were also identified during literature screening (section 2.2).

2.2 Screening of literature

A two-step approach was adopted to screen primary literature for inclusion in the risk assessment. In an initial screen, titles and abstracts were reviewed to assess their relevance. For those papers considered to be relevant, or potentially relevant, full papers were obtained and evaluated in a second screening step to confirm their relevance and assess scientific


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robustness. Klimisch scoring of primary literature was not required due to the lack of relevant studies being identified (see section 3.1).

2.3 Risk assessment process

Risk assessment is traditionally taken to comprise four elements:

• Hazard identification

• Dose-response assessment

• Exposure assessment

• Risk characterisation The risk assessment outlined here refers to the process in which the potential for adverse health effects following exposure of humans to identified chemicals in PCPs and DCPs in drinking water was evaluated. Three age groups were considered, adults, children and infants, to highlight any potential differences that might arise as a consequence of body weight.

2.3.1 Hazard identification

During this step, an evaluation was made for each of the chemicals and their potential adverse health effects. This was identified through the review of toxicological data, with the following weighting: human epidemiology data > experimental data > in vitro data > QSAR modelling.

2.3.2 Dose-response assessment

Also referred to as ‘hazard characterisation’, this step determined how much of each chemical would be required to cause an adverse effect, and what level of exposure would result in negligible or non-existent effects. This was achieved as detailed below.

2.3.2.1 Identification of a relevant POD

The chemicals evaluated here were all assessed as having a threshold of toxicity, i.e. a dose could be defined (a POD) below which toxicity did not occur. PODs typically used for risk assessment purposes include the No Observed Adverse Effect Level (NOAEL), the Lowest Observed Adverse Effects Level (LOAEL) and the benchmark dose (BMD). The availability of these PODs for each chemical under consideration was identified from the collated toxicity data (section 2.1) or through derivation, as required, on a case-by-case basis.

The NOAEL is defined as the highest dose at which no adverse effects are seen in a toxicity study. If a NOAEL cannot be determined from the data, due to effects being seen at even the lowest dose tested, the LOAEL i.e. the lowest dose at which some adverse effects are seen may be determined. Use of the NOAEL or LOAEL as a POD for risk assessment purposes has some recognised limitations, as these must correspond to one of the doses used in the study, which can be just a few doses in a wide range. Therefore, the true “no effect level” could conceivably be higher than the experimental NOAEL, depending on the sensitivity of the study and the choice of endpoint. This makes the NOAEL a less informative value in some studies but remains preferable to the use of a LOAEL for risk assessment purposes. An alternative POD, the BMD, was collated where available. This is the dose that produces a predetermined change in response (known as the benchmark response (BMR)) for a given toxicological effect. For risk assessment purposes, the 95 % lower confidence limit of the BMD (BMDL) is often


15

used as the POD. The use of the BMD is considered to be beneficial as it is based on all available data on the dose response and is on the scale of observable effects rather than being based on one somewhat uncertain data point such as the NOAEL (EFSA, 2017).

2.3.2.2 HBGV identification

A HBGV represents the dose that is protective of the general population, including sensitive subpopulations, and is defined as ‘the estimated dose in humans that can be ingested over a lifetime without appreciable risk to health’. Typical HBGVs include the acceptable daily intake (ADI), tolerable daily intake (TDI) or, in some cases, a tolerable weekly intake (TWI) which are expressed as mg/kg bw/day. These were either identified as part of the literature screening process, or if not available, calculated using the equation (WH O, 2017):

𝐻𝐵𝐺𝑉 = 𝑃𝑂𝐷 ÷ 𝐴𝐹

Where a number of PODs were identified, the one relevant to the most sensitive toxicological endpoint was utilised to calculate the HBGV, unless otherwise specified.

When deriving a HBGV using a NOAEL from a chronic animal study, a default Assessment Factor (AF), also previously known as uncertainty factor (UF), of 100 is typically applied (EFSA, 2012). This takes into consideration a factor of 10 to account for interspecies variability (4 for toxicokinetics and 2.5 for toxicodynamics) and 10 to account for intraspecies differences (3.2 for toxicokinetics and 3.2 for toxicodynamics). If a LOAEL is used an additional AF is applied (EFSA, 2012). Other AFs may be used depending on the type, quality and outcome of the toxicity study, as detailed in Table 2.

2.3.3 Exposure assessment

This step determined how much of each chemical a person could be exposed to via drinking water, using either measured values (where available) or the maximum drinking water levels estimated in the previous research study (WRc, 2014) as a worst-case scenario where measured data is unavailable. Default parameters for body weight and volume of water ingested were used to estimate intake of each chemical on a body weight basis for three receptors (World Health Organisation, 2017). It should be noted that these levels of consumption are expected to be conservative in nature.

• 60 kg adult drinking 2 L per day

• 10 kg child drinking 1 L per day

• 5 kg infant drinking 0.75 L per day

2.3.4 Risk characterisation

During this step, relevant information gathered in the preceding steps was integrated to characterise the risk to humans following exposure to each chemical via drinking water. The intakes calculated in step 2.3.3 were compared to the calculated HBGV to determine the hazard quotient (HQ). Where the HQ is <1 no adverse effects are expected to occur; if the HQ is > 1 it is considered possible that adverse effects may occur.


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Table 2: Assessment factor considerations

AF Possible Range Purpose

Interspecies differences 1-10

Accounts for differences in sensitivity between species. 10 is proposed if animal data are used and 1 is proposed if human data is used.

Intraspecies differences 1-10

Accounts for differences in sensitivity between individuals. 3 is proposed if human data are used, depending on the sub-population and 3 is proposed if using a therapeutic dose.

Conversion of LOAEL to NOAEL

3-10

Accounts for the test dose causing some adverse effect. 10 is proposed when NOAEL is not available and 10 is proposed if the LOAEL is based on a therapeutic dose.

Use of subchronic exposure data

1-10

Accounts for the test study being less than chronic exposure. 10 is proposed when no chronic data are available; 1 is proposed when chronic data are used; 3 is proposed for epidemiology studies or developmental studies.

Inadequate databases

1-10

Used to account for less than adequate datasets. 3 is proposed if QSARs are used and 3 is proposed if the NOAEL is the highest dose tested.

Source: ECHA, 2012

A further approach that is often used to indicate the level of concern for an exposure scenario is the margin of exposure (MOE). This was calculated for each chemical as the ratio of the NOAEL or BMD for the critical effect to the estimated exposure. It was considered that the larger the MOE, the smaller the potential risk following exposure to the chemical (Benford et al., 2010). For non-carcinogenic substances, MOE’s of >100 were considered acceptable when derived from NOAELs or BMDs (US Environmental Protection Agency, 2012).

The HBGVs (TDIs) were also used to calculate drinking water equivalent levels (DWEL), using the default body weight (bw) and intake exposure parameters described above and a 20 % allocation (P) to drinking water (US EPA, 2018):

𝐷𝑊𝐸𝐿 = 𝑇𝐷𝐼 × 𝑏𝑤 × 𝑃 ÷ 𝑖𝑛𝑡𝑎𝑘𝑒 𝑣𝑜𝑙𝑢𝑚𝑒

For each chemical, the DWEL was compared to levels measured in drinking water, if available, or the maximum modelled concentrations, both previously reported (WRc, 2014). Where the DWEL was greater than the reported levels, it was concluded that there is no risk to public health following ingestion via drinking water.

General references

Benford D, Bolger PM, Carthew P, Coulet M, DiNovi M, Leblanc JC, Renwick AG, Setzer W, Schlatter J, Smith B, Slob W, Williams G and Wildemann T, 2010. Application of the Margin of


17

Exposure (MOE) approach to substances in food that are genotoxic and carcinogenic. Food Chem Toxicol, 48 Suppl 1, S2-24.

Bull S., Green O., Carter J. (2014) Toxicological evaluation for pharmaceuticals in drinking water. Report to DWI. Ricardo-AEA Ltd. Ref: ED59005- Issue Number 2. Available at: http://dwi.defra.gov.uk/research/completed-research/reports/DWI70-2-295.pdf [accessed July 2020].

ECHA (2012) European Chemicals Agency. Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health. European Chemicals Agency. Available at: https://www.echa.europa.eu/documents/10162/13632/information_requirements_r8_en.pdf [accessed September 2020].

ECHA (2016) European Chemicals Agency. Guidance on Information Requirements and Chemical Safety Assessment Part E: Risk Characterisation Version 3.0 May 2016. Available at: https://echa.europa.eu/documents/10162/13632/information_requirements_part_e_en.pdf/1da6cadd-895a-46f0-884b-00307c0438fd [accessed September 2020].

US EPA (2012) US Environmental Protection Agency. Sustainable futures. P2 framework manual - Quantitative Risk Assessment Calculations. EPA-748-B12-001. Available at: https://www.epa.gov/sustainable-futures/sustainable-futures-p2-framework-manual [accessed July 2020].

US EPA (2018) US Environmental Protection Agency. 2018 Edition of the Drinking Water Standards and Health Advisories. EPA 822-F-18-001. Available at: https://www.epa.gov/sites/production/files/2018-03/documents/dwtable2018.pdf [accessed September 2020].

WHO (2017) World Health Organisation Guidelines for drinking-water quality: fourth edition incorporating the first addendum. Geneva: World Health Organization; 2017. Licence: CC BY-NC-SA 3.0 IGO. Available at: https://apps.who.int/iris/bitstream/handle/10665/254637/9789241549950-eng.pdf;jsessionid=24C9F5F9C7ED9014D59219ACC5B43346?sequence=1 [accessed July 2020].

WRc (2014). Risk to drinking water from Personal Care Products and Domestic Cleaning Products. Available at: http://dwi.defra.gov.uk/research/completed-research/reports/DWI70-2-283.pdf [accessed Jan 2020].

http://dwi.defra.gov.uk/research/completed-research/reports/DWI70-2-295.pdf

https://www.echa.europa.eu/documents/10162/13632/information_requirements_r8_en.pdf

https://www.echa.europa.eu/documents/10162/13632/information_requirements_r8_en.pdf

https://echa.europa.eu/documents/10162/13632/information_requirements_part_e_en.pdf/1da6cadd-895a-46f0-884b-00307c0438fd

https://echa.europa.eu/documents/10162/13632/information_requirements_part_e_en.pdf/1da6cadd-895a-46f0-884b-00307c0438fd

https://www.epa.gov/sustainable-futures/sustainable-futures-p2-framework-manual

https://www.epa.gov/sites/production/files/2018-03/documents/dwtable2018.pdf

https://apps.who.int/iris/bitstream/handle/10665/254637/9789241549950-eng.pdf;jsessionid=24C9F5F9C7ED9014D59219ACC5B43346?sequence=1

https://apps.who.int/iris/bitstream/handle/10665/254637/9789241549950-eng.pdf;jsessionid=24C9F5F9C7ED9014D59219ACC5B43346?sequence=1




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3.0 Results

3.1 Identification of data

Relevant data for each chemical were collated through a search of published papers and grey literature, as described in section 2.1. A two-step screening process was then adopted, as described in section 2.2, to identify data that could be specifically used for risk assessment purposes. Table 3 provides a summary of the number of papers identified at each stage of primary literature screening, and of the number of sources of grey literature available for each chemical of interest.

Table 3: Primary and secondary screening of identified literature

Chemical name CAS No. Number of papers retrieved

Number following primary screening

Number following secondary screening

Grey Literature identified

1,2,3-benzotriazole 95-14-7 54 2 0 7 sources

1H-benzotriazole, 4(or 5)-methyl

29385-43-1 24 0 n/a 5 sources

(1-hydroxyethylidene) diphosphonic acid (HEDP)

2809-21-4

173 2 0 11 sources

Diethylenetriamine penta(methylene phosphoric acid) (DTPMP)

15827-60-8

13 0 n/a 9 sources

Amino tris(methylene phosphoric acid) (ATMP)

6419-19-8 21 0 n/a 10 sources

2-(2-butoxyethoxy) ethanol (DEGBE)

112-34-5 15 7 0 24 sources

Linear alkylbenzene sulphonate (LAS)

68411-30-3 92 7 0 9 sources (LAS)

10 sources (SAS)

Cocamidopropyl betaine (cosmetic grade) Cocamidopropyl betaine (technical grade)

61789-40-0; 83138-08-3; 86438-79-1 4292-10-8

23 7 0 12 sources

Although primary literature was identified for all chemicals, following the screening process none were considered useful for risk assessment purposes (Annex B). The following sections provide a detailed step-by-step risk assessment for each of the chemicals of interest which have been based on information collated from the grey literature identified during initial searches.


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3.2 Human health risk assessments for exposure via

drinking water

The following section (A – H) presents the individual chemical assessments for the 8

separate chemicals as follows:

Section A 1,2,3-benzotriazole

Section B 1H-Benzotriazole, 4(or 5)-methyl-

Section C (1-hydroxyethylidene) diphosphonic acid (HEDP)

Section D Diethylenetriamine penta (methylene phosphonic acid) (DTPMP)

Section E Amino tris(methylene phosphonic acid) (ATMP)

Section F 2-(2-butoxyethoxy) ethanol (DEGBE)

Section G Linear alkylbenzene sulphonate (LAS)

Section H Cocamidopropyl betaine (CAPB)


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Section A: 1,2,3-benzotriazole

Section A1.0 Chemical identification, use and

potential human exposure

Section A1.1 Reasons for consideration

WRc, (2014) previously assessed the risk to reach drinking water following use of 1,2,3-benzotriazole as a corrosion inhibitor in dishwasher detergents. In the UK, consumption decreased from 832 tonnes/year in 2007 to 390 tonnes/year in 2012 (WRc, 2014). Levels of 1,2,3-benzotriazole in drinking water were measured as 0.79 µg/L and estimated to be in the range 3.77 – 60 µg/L; the authors noted that this was based on estimated effluent concentrations up to sixteen times higher than measured concentrations reported in the literature. 1,2,3-benzotriazole is included for further consideration here as the estimated exposures through drinking water and bathing were found to be greater than the estimated exposure (calculated as Systemic Exposure Dose or SED; 0.00002 mg/kg bw/day) through intended use at the maximum predicted levels (WRc, 2014).

Section A1.2 Identification and physicochemical properties.

1,2,3-benzotriazole is a heterocyclic compound, existing in two tautomeric forms, 1H-benzotriazole and 2H-benzotriazole. The more stable of these is 1H-benzotriazole and is essentially the exclusive molecular structure, presented in Table A1.1.

Section A1.3 Hazard classifications

1,2,3-benzotriazole has been assigned a hazard category of Acute Tox 4, with hazard statement H302 – “harmful if swallowed”, and Eye Irrit.2, with hazard statement H319 – “causes serious eye irritation” in the REACH dissemination page4.

Notified EU classification and labelling, according to CLP regulation (EC) No. 1272/2008, includes Acute Tox. 4 with hazard 302 “harmful if swallowed”. There is no harmonised classification.

Section A1.4 Occurrence, production and use.

Benzotriazole is produced by reaction of o-phenylenediamine with nitrous acid in the presence of glacial acetic acid or by reaction of hydrochloric acid or nitrous acid with o-phenylenediamine. Benzotriazole is used as a corrosion inhibitor, as a plastic stabiliser, and as a chemical intermediate for dyes, pharmaceuticals and fungicides (Stouten et al., 2000). As detailed above, the annual production of 1,2,3-benzodiazole in the EU in 2012 was 390 tonnes. No information relating to the levels used in dishwasher products was identified in the public domain.

4 https://echa.europa.eu/information-on-chemicals/cl-inventory-database/-/discli/details/36314

https://echa.europa.eu/information-on-chemicals/cl-inventory-database/-/discli/details/36314


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Table A1.1: 1,2,3-benzotriazole identification and physicochemical properties

Parameter Specification

Cas No

95-14-7

IUPAC name 1,2,3-benzotriazole

Chemical Group Heterocyclic

Physical form Crystalline powder

Domestic Substances List (DSL) name

Not on list

Common name 1-(diphenylmethyl)-1H-1,2,3-benzotriazole

Formula: C6H5N3

Molecular weight 119.14

Structure

Solubility (water) 1.98 x 104mg/L

Melting point 100oC

Boiling point 350oC

Density 1.36 g/cm3

Log Pow 1.34

Vapour Press. 2.40 x 10-5 mm Hg

Sources: WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for 1,2,3-benzotriazole.

Section A1.5 Human exposure

Occupational exposure is the most likely source of exposure to 1,2,3-benzotriazole in humans, with dermal and inhalation being key routes. Non-occupational exposure may occur from the use of DCPs or through ingestion of drinking water that has been contaminated with 1,2,3-benzotriazole (WRc, 2014).

Section A2.0 Human relevant health effects

Section A2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of 1,2,3-benzotriazole in humans or experimental species following exposure via any route.

In the absence of data, the REACH registration document utilises the physicochemical properties of 1,2,3-benzotriazole to estimate its toxicokinetics (ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020). Absorption via the oral and dermal route were assumed to be 100 %, and via inhalation to be 10 %. Bioaccumulation potential was also considered to be low. An in vitro study in which rat liver microsomes were incubated for one-hour with 1,2,3-benzotriazole, suggests that metabolism to 4- and 5-hydroxybenzotriazole takes place, but at a low rate. Elimination was estimated to occur through the urine and faeces (Beltoft et al., 2013).


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Section A2.2 Acute toxicity

No data was identified relating to the acute effects of 1,2,3-benzotriazole in humans via any route of exposure.

The acute oral toxicity of 1,2,3-benzotriazole has been tested in rats, mice and the guinea pig. Acute oral toxicity studies (carried out prior to OECD/GLP standards) have reported LD50 values in rats between 500 and 965 mg/kg bw. In mice, oral LD50 values ranged between 615 and 831 mg/kg bw (one study also reported a value >4500 mg/kg bw, however the strain was unspecified). A single oral LD50 value of 500 mg/kg bw has been reported in the guinea pig (Beltoft et al., 2013; DECOS, 2000).

Sublethal effects were noted in three studies in which rats which were administered 1,2,3-benzotriazole by oral gavage at doses between 46.4 and 2250 mg/kg bw. Non-lethal clinical signs of toxicity included depressed righting and placement reflexes, absence of pain, shallow respiration, temporary prostration and lethargy (Beltoft et al., 2013; DECOS, 2000).

Based on the acute lethal toxicity data and using EC-classification criteria, 1,2,3-benzotriazole is considered to be harmful following oral exposure (DECOS, 2000).

Section A2.3 Repeat dose toxicity

Section A2.3.1 Systemic effects

No data were identified relating to the systemic effects of 1,2,3-benzotriazole in humans via any route of exposure.

The effects of dietary exposure of F344 rats and B6C3F1 mice to 1,2,3-benzotriazole over an 8-week period were reported by NTP (1978) in a dose ranging study for the bioassay detailed below. Rats and mice were administered doses of 0, 300, 1000, 3000, 10,000 and 30,000 ppm (equivalent to 0, 15, 50, 150, 500 and 1500 mg/kg bw/day in rats and 0, 45, 150, 450, 1500 or 4500 mg/kg bw/day in mice). Mean body weights were [statistically] significantly decreased (no p value given) in rats at the highest dose when compared to controls, in both males and females. Low and high doses of 10,000 and 20,000ppm (500 and 1000 mg/kg bw/day respectively) were identified for use in the chronic study for rats. In mice, no [statistically] significant (no p value given) decrease in mean body weight was noted in males or females for any dose group. Low and high doses of 20,000 and 40,000 ppm (300 and 600 mg/kg bw/day) were identified for use in the chronic study in mice. No further details regarding systemic effects following subchronic exposure to 1,2,3-benzotriazole in rats or mice was provided (NTP, 1978).

In a subsequent long-term oral study conducted according to NTP protocols (similar to OECD Guideline 451 – Carcinogenicity Studies) Fischer 344 rats were administered 1,2,3-benzotriazole at doses of 0, 6,700 or 12,100 ppm (0, 305 or 605 mg/kg bw/day) for 78 weeks followed by a 26 week observation period (NTP, 1978). Survival rates were unaffected in males and slightly higher than controls in treated females (p=0.049). Growth curves showed that exposed groups had lower mean body weights than controls. Exposure to 1,2,3-Benzotriazole at either dose was not associated with clinical signs of toxicity. Histopathology indicated the presence of cellular effects in a number of organs including: liver, kidney,


23

pancreas, lung, prostate, uterus and ovary at both doses, but in a non-dose dependent manner (NTP, 1978). A LOAEL of 305 mg/kg bw/day can be derived from the study based on body weight changes.

A long-term oral study was also carried out in B6C3F1 mice (50 animals/sex/group) according to NTP protocols. 1,2,3-Benzotriazole was administered in the diet at levels of 11,700 and 23,500 ppm (equivalent to 585 and 1175 mg/kg bw/day) for 104 weeks with a further 2 week observation period (NTP, 1978). Survival rates were unaffected in male mice and higher than controls (p=0.002) in treated females at the highest dose. Growth curves showed a dose-related [statistically] significant (no p value given) decrease in body weights when compared to controls in both sexes. Exposure of B6C3F1 mice to 1,2,3-benzotriazole at either dose was not associated with clinical signs of toxicity. Histopathological changes were observed in bone marrow, lymph nodes, kidney, spleen and lungs at both doses in a non-dose dependent manner (NTP, 1978). A LOAEL of 11755 mg/kg bw/day can be derived based on body weight changes.

Section A2.3.2 Irritancy and corrosivity

No data was identified relating to the irritancy potential of 1,2,3-benzotriazole in humans (see also section A2.3.3).

In a study carried out to OECD Guideline 404 (Acute Dermal Irritation/Corrosion), 1,2,3-benzotriazole did not cause skin irritation in the rabbit at a dose of 500 mg when applied under semi-occlusive conditions for 4 h to intact clipped skin (observation up to 14 days post exposure) and is deemed not to meet the CLP requirements for classification as a skin irritant (ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020).

An acute eye irritation study carried out to OECD Guideline 405 (Acute Eye Irritation/Corrosion) was reported in the rabbit in which 1,2,3-benzotriazole resulted in moderate irritation. As such, it meets the criteria for classification under CLP regulation in Category 2 (ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020).

Section A2.3.3 Sensitisation

Data relating to the sensitising potential of 1,2,3-benzotriazole in humans provides equivocal evidence. In workers with contact dermatitis who had been exposed occupationally to benzotriazole-containing oils and greases, positive reactions (weak to strongly positive) were seen in a patch test of 2 % benzotriazole (in petrolatum) in 4 of the individuals (total number of subjects not known). However, the study is limited for interpretation as it did not report data on negative controls or whether an immunological response had occurred. A further study in Dutch metalworkers with contact dermatitis did not report positive patch tests when challenged with benzotriazole (ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020).

Based on the available data for humans, an irritating and/or sensitizing potential of 1,2,3-benzotriazole cannot be excluded.

1,2,3-benzotriazole was reported to be a non sensitising agent in a guinea pig maximisation test carried out to OECD Guideline 406 (Skin Sensitisation) (Bingham et al., 2001 – cited in ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020).


24

Section A2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of 1,2,3-benzotriazole in humans via any route of exposure.

In vitro studies have also been carried out to assess the mutagenic potential of 1,2,3-benzotriazole at doses between 444 and 8000 µg/0.1 mL. Positive results were reported in S. typhimurium strain TA1535 in the presence of a metabolic activation system (Ciba-Geigy Corp, 1993; Dunkel et al., 1985; Zeiger et al., 1987 – all cited in Stouten et al., 2000). Positive (Ciba-Geigy Corp, 1993) and negative (Dunkel et al., 1985; Zeiger et al., 1987 – all cited in Stouten et al., 2000) outcomes have been reported in TA1535 in the absence of metabolic activation. Dunkel et al., (1980) reported that a positive response was only seen in the presence of S9 from hamster liver, and not S9 derived from mouse and rat liver. Negative findings have been reported for 1,2,3-benzotriazole tested in TA97, TA98, TA100, TA1537, TA1538 both in the presence and absence of S9 (Dunkel et al., 1980; Dunkel et al., 1985; Zeiger et al., 1987; Ciba-Geigy Corp, 1993 – all cited in Stouten et al., 2000). Commercial benzotriazole (purity not reported) gave positive results when tested in strains TA98, TA1537, and TA1538 in the presence and absence of S9 (Ciba-Geigy Corp, 1993 – cited in Stouten et al., 2000).

1,2,3-benzotriazole was also positive for mutagenicity in E. coli (strain WP2 uvrA) under both metabolic conditions (Dunkel et al., 1985 – cited in Stouten et al., 2000). An indication test for DNA damage (SOS chromotest in E. coli PQ37) was negative at doses up to 100 mM (no further details given) (Beltoft et al., 2013; DECOS, 2000).

1,2,3-benzotriazole has been tested in the HPRT forward mutation assay in CHO cells according to OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test using Hprt and Xprt genes) and was negative in the absence of S9 metabolic activation at doses between 400 and 1000 µg/ml (Den Boer, 1987 – cited in Stouten et al., 2000).

The cytogenic potential of 1,2,3-benzotriazole has been assessed in an in vivo study carried out to OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test). A single oral gavage dose of 800 mg/kg/bw (previously shown to cause toxicity) administered to male and female mice was not found to be associated with an increase in the incidence of micronucleated polychromatic erythrocytes at either 24, 48 or 72 h post exposure (DECOS, 2000).

It should be noted that the REACH Registration document does not include consideration of all of the published in vitro study findings for mutagenicity and concludes from the studies performed by industry that there is no evidence for genotoxicity (ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020).

Overall, considering the weight of evidence from the genotoxicity data package, the greatest weight can be given to the in vivo micronucleus test (OECD guideline 474) which was favourable. In vitro data from a number of types of study have shown equivocal results.

Section A2.3.5 Carcinogenicity

No data relating to the carcinogenic potential of 1,2,3-benzotriazole in humans were identified.


25

A carcinogenicity study was carried out according to NTP protocols (similar to OECD Guideline 451 – Carcinogenicity Studies) (NTP, 1978). Fischer 344 rats were administered 1,2,3-benzotriazole in the diet at doses of 6,700 or 12,100 ppm (equivalent to 305 and 605 mg/kg bw/day) for 78 weeks, followed by a 26-week observation period. Systemic toxicity findings are detailed in section 2.3.1. Male rats presented with neoplastic nodules in the liver, the incidence of which was statistically significantly (p=0.024) higher in the high dose group than in controls (5/45 and 0/45; 11 and 0 % respectively). However, the incidence was found to be within the range (0 to 11 %) for historical control data at the laboratory and not considered to be treatment related. Brain tumours were noted in some males at the lowest dose (one oligodendroglioma, two gliomas) and in one female at the highest dose (glioma). These were interpreted by the authors as being suggestive, but not sufficient, evidence of carcinogenicity. A significant increase (p=0.01) in incidence of endometrial stromal polyps in female rats at the low dose was judged as not treatment related by the authors, as incidence in the high dose group was not significantly changed. Similarly, an increase in the incidence of thyroid C-cell adenomas and carcinomas over that of controls was seen in low dose females (5/43 compared to 0/43; 12 and 0 %) - higher than the number seen at the highest dose (3/50; 6 %). Only benign thyroid tumours occurred at the low dose of 1,2,3-benzotriazole (4/43; 9 %), while malignant thyroid tumours were seen in both groups (1/43 and 3/50 at low and high doses; 2 and 6 %, respectively). The authors reported that although incidence appeared to be dose related, the rates were comparable to incidences reported in untreated females from other laboratories and not considered to be treatment related (NTP, 1978).

A carcinogenicity study in B6C3F1 mice (50 animals/sex/group) has also been carried out according to NTP protocols (NTP, 1978). 1,2,3-benzotriazole was administered in the diet at levels of 11,700 and 23,500 ppm (equivalent to 585 and 1755 mg/kg bw/day) for 104 weeks with a further 2-week observation period. No tumours were observed in male mice at incidences above those in control groups. In female mice, a statistically significant increase (p=0.001) in the incidence of alveolar/bronchiolar carcinomas over that in controls was seen at the low dose (9/49 compared with 0/49; 18 % and 0 %). However, this was judged by the authors as not being treatment related as incidence in the high dose group was not significantly changed from controls (3/49 compared with 0/49; 6 % and 0 %). In addition, the incidence of the tumours in historical controls was comparable to that seen at the highest dose (between 0 and 7 %). The evidence was described as being suggestive, but not sufficient evidence of a carcinogenic effect, as reported for rats above (NTP, 1978).

The authors of the carcinogenicity studies concluded that, under the conditions of the bioassay, there was no convincing evidence that 1,2,3-benzotriazole was carcinogenic in F344 rats or B6C3F1 mice of either sex (NTP, 1978). The consensus view in the available hazard assessments by authoritative bodies (DECOS, 2000; Beltoft et al., 2013; Stouten et al., 2000) is that the database (both for mutagenicity and carcinogenicity endpoints) is inadequate to allow classification of 1,2,3-benzotriazole as to its carcinogenic potential.

Section A2.3.6 Reproductive and developmental toxicity

No data relating to the reproductive and/or developmental toxicity of 1,2,3-benzotriazole in humans were identified via any route of exposure.


26

The REACH registration document reports an unnamed reproductive toxicity study carried out in 2012 to OECD Guideline 421 (Reproduction/Developmental Toxicity Screening Test) in which 1,2,3-benzotriazole was administered by oral gavage to male and female Wistar rats at doses of 0, 12.5, 50 and 200 mg/kg bw/day. No clinical or general signs of toxicity were apparent and there was no indication that reproductive performance had been adversely affected in either sex. Histopathology of the ovaries and epididymides/testes was also unremarkable. No developmental toxicity was evident in the offspring of exposed parents. It is noted by the registrant that OECD Guideline 421 does not provide complete information concerning all aspects of reproduction and development or post-natal effects that may become apparent due to pre-natal exposure (ECHA REACH registration dossier for 1,2,3-benzotriazole, 2020).

Section A2.3.7 Specific considerations

1,2,3-benzotriazole is currently being investigated as a potential environmental endocrine disruptor. ECHA state that ‘there is scientific evidence from in vitro as well as in vivo studies, that 1-h-benzotriazole can bind to the oestrogen receptor and act as oestrogen agonist, leading to adverse effects in organisms in the environment. UBA considers a substance evaluation for 1-h-benzotriazole as necessary to check if the concerns regarding endocrine disrupting properties are sufficient to confirm that it is as endocrine disrupting substance for the environment”5.

Section A2.3.8 Summary of human relevant health effects

• No information relating to the potential acute toxicity of 1,2,3-benzotriazole in humans was identified.

• Toxicokinetic data are absent for both humans and experimental species. Bioaccumulation potential has been estimated as low and excretion is considered to occur via urine and faeces. In vitro evidence suggests that metabolism to 4- and 5-hydroxybenzotriazole takes place in the liver, but at a low rate.

• According to EC-classification criteria, 1,2,3-benzotriazole is considered harmful via the oral route.

• Based on the available data for humans, an irritating and/or sensitising potential of 1,2,3-benzotriazole cannot be excluded.

• In vivo, benzotriazole was negative in an oral mouse bone marrow micronucleus assay, and this study carries the most weight. In vitro mutagenicity testing provides variable findings. In vitro, 1,2,3-benzotriazole is mutagenic in Salmonella typhimurium TA 1535 and in E. coli, but not in CHO cells. The SOS chromotest in E. coli, an indicator test for DNA damage was negative.

• There is inconclusive evidence from carcinogenicity studies with rats and mice as to the carcinogenic potential of 1,2,3-benzotriazole. The studies are old and the database inconclusive to support a definitive conclusion regarding carcinogenicity of this chemical.

• Repeat dose oral studies showed adverse effects of 1,2,3-benzotriazole on body weight. Due to poorly reported data, it is not possible to determine if this is a significant effect.

5 https://echa.europa.eu/substance-information/-/substanceinfo/100.002.177

https://echa.europa.eu/substance-information/-/substanceinfo/100.002.177


27

• Reproductive and developmental toxicity studies did not show adverse effects following oral exposure to 1,2,3-benzotriazole.

Section A3.0 Identification of relevant point(s) of

departure for use in risk assessment

Section A3.1 Critical endpoint for human health risk assessment

purposes

The POD for use in the risk assessment (see Section A4.0) should, where possible, be derived from a repeated dose toxicity study, preferably using the oral route of exposure. There are no human studies relating to toxicological effects following exposure to 1,2,3-benzotriazole through any exposure route. However, a number of oral repeated dose experimental studies have been carried out for 1,2,3-benzotriazole, for systemic and specific organ toxicity. These are summarised in Table A3.1.


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Table A3.1 Oral route toxicity studies for 1,2,3-benzotriazole

Type/Duration Species/Strain Dose range (mg/kg bw/ day); route of exposure

Critical effects / Comments Reference

Repeated dose 8 weeks

Rat / Fischer 344 15 – 1500 via the diet

At the highest dose – mean body weights decreased in males and females. Low and high doses of 500 and 1000 mg/kg bw/day identified for use in the bioassay.

NTP 1978


Mice / B6C3F1 45 – 4500 via the diet

No decrease in mean body weight at any dose in males or females. Low and high doses of 300 and 600 mg/kg bw/day identified for use in the bioassay.

NTP 1978


Rat / Fischer 344 305 or 605 via the diet

Lower body weight at both doses when compared to controls in both males and females. Non-dose dependent histopathological changes in liver, kidney, pancreas, lung, prostate, uterus and ovary at both doses. Observed neoplastic changes were not considered as treatment related. A LOAEL of 305 mg/kg bw/day was derived from the study based on body weight changes.

NTP 1978


Mice / B6C3F1 1755 or 3525 via the diet

Dose related decrease in body weight in males and females. Non-dose dependent histopathological changes in bone marrow, lymph nodes, kidney, spleen and lungs, at both doses. Observed neoplastic changes were not considered as treatment related. A LOAEL of 1755 mg/kg bw/day was derived based on body weight changes.

NTP 1978

OECD 421 Reproduction/ Developmental

Rat / Wistar 12.5 – 200 by oral gavage

No treatment related effects on reproductive performance in either sex at any dose.

ECHA REACH registration dossier for 1,2,3-


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Toxicity Screening Test

No developmental toxicity apparent at any dose. A NOAEL of 200 mg/kg bw/day (highest dose tested) was derived for reproductive and developmental toxicity.

benzotriazole, 2020; unnamed study

Shaded rows represent studies considered suitable for use in the risk assessment


30

The data indicate that the lowest POD identified is the NOAEL from the Reproduction / Developmental Toxicity Screening Test following oral exposure to 1,2,3-benzotriazole. There is no evidence from currently available studies that 1,2,3-benzotriazole is carcinogenic (discussed further in section A3.3).

Section A3.2 Current guideline values

No health-based guidance values are available.

Section A3.3 Identification of POD to be used for the risk

assessment

A LOAEL of 305 mg/kg bw/day was determined in a chronic duration study in rats, however the most sensitive of the PODs identified for 1,2,3-benzotriazole is the NOAEL of 200 mg/kg bw/day derived for reproductive/developmental toxicity. This is based on the highest dose tested. Both POD are considered appropriate for use in risk assessment.

Section A4.0 – Drinking water risk assessment

Hazard identification

Few adverse effects were seen following oral exposure to 1,2,3-benzotriazole in experimental studies.

Hazard characterisation

Based on the data obtained, a LOAEL of 305 mg/kg bw/day and a NOAEL of 200 mg/kg bw/day are considered appropriate to use as PODs.

Assessment factors (AF)

An AF of 1000 is considered appropriate for use with the LOAEL from the chronic study and an AF of 600 for use with the NOAEL from the Reproduction / Developmental Toxicity Screening Test.

LOAEL NOAEL POD (mg/kg bw/day) 305 200 AF AF Justification

Interspecies differences 10 10 POD based on animal data

Intraspecies differences 10 10 To account for human variability

Use of subchronic data 1 3 NOAEL based on a < 90 day study

Inadequate databases 1 2 Inconclusive carcinogenicity database.

Conversion of LOAEL to NOAEL 10 1 Only LOAEL available

Total AF 1000 600

Derivation of the TDI

Based on the LOAEL of 305 mg/kg bw/day and using an AF of 1000, the proposed TDI is 305 µg/kg bw/day. Based on the NOAEL of 200 mg/kg bw/day and using an AF of 600, the proposed TDI is 333 µg/kg bw/day. This shows that both POD give comparable TDIs, and 305 µg/kg bw/day is selected for risk assessment purposes as this is the most precautionary in nature.


31

Derivation of drinking water equivalent level (DWEL)

Based on a TDI of 305 µg/kg bw/day, and assuming 20 % allocation of the TDI to drinking water, the DWELs would be:

• 1830 µg/L for a 60 kg adult drinking 2 L water per day

• 610 µg/L bw/day for a 10 kg child drinking 1 L water per day

• 407 µg/L bw/day for a 5 kg infant drinking 0.75 L water per day

Exposure assessment

The maximum concentration of 1,2,3-benzotriazole measured in drinking water, as reported by WRc (2014), was 0.79 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.

Based on default assumptions, the daily intake at this concentration would be:

• 0.030 µg /kg bw/day for a 60kg adult drinking 2L a day

• 0.079 µg /kg bw/day for a 10kg child drinking 1L a day

• 0.12 µg /kg bw/day for a 5kg infant drinking 0.75L a day This assumes that the total exposure to 1,2,3-benzotraizole is via the consumption of drinking water however it is feasible that other intakes may occur during use of the product.

Risk characterisation

The maximum intake of 1,2,3-benzotriazole via drinking water by adults, children and infants is less than the TDI (HQs <1) and the measured concentration of 1,2,3-benzotriazole in drinking water is less than the DWEL. No special sensitivity to the chemical was identified in any age group from the available literature. Therefore, no adverse public health effects are anticipated following exposure to 1,2,3-benzotriazole via drinking water.

Risk communication

The MOEs for 1,2,3-benzotriazole, based on the LOAEL of 305 mg/kg bw/day and measured intake are 11582278, 3860759 and 2573840 for adults, children and infants, respectively. As MOEs are > 100 this indicates that the anticipated exposures are not of concern in terms of risk to public health.

References

Beltoft V., Nielsen E., Ladefoged O. (2013) - The Danish Environmental Protection Agency. 1526 Benzotriazole and Tolyltriazole. Evaluation of health hazards and proposal of health based quality criteria for soil and drinking water. Available at: https://www2.mst.dk/Udgiv/publications/2013/12/978-87-93026-81-0.pdf [accessed June 2020].

Bingham E., Cohrssen B., Powell CH. Patty's Toxicology Volumes 1-9 5th ed. John Wiley & Sons. New York, N.Y. (2001)., p. V4 1178.

https://www2.mst.dk/Udgiv/publications/2013/12/978-87-93026-81-0.pdf


32

Ciba-Geigy Corp. Salmonella/mammalian-microsome mutagenicity test with TK 10´637 with cover letter dated 072893. Springfield VA, USA: National Technical Information Service (NTIS), 1993; order no NTIS/OTS0538213 (cited in Stouten et al., 2000).

Den Boer WC. Mutagenicity test on Benzotriazol Granulat in the CHO HGPRT forward mutation assay. Veenendaal, the Netherlands: Hazleton Biotechnologies Veenendaal Laboratory, 1987; HBC study no E-95553-0-435 (study submitted to Bayer AG, Institut für Toxikologie, Wuppertal, Germany) (unpublished data submitted to DECOS by Bayer AG) (cited in Stouten et al., 2000).

Dunkel VC, Simmon VF. Mutagenic acitivity of chemicals previously tested for carcinogenicity in the national cancer institute bioassay program. IARC Sci Publ 1980;27: 283-302. 16 (cited in Stouten et al., 2000).

Dunkel VC, Zeiger E, Brusick D, McCoy E, McGregor D, Mortelmans K, Rosenkranz HS, Simmon VF. Reproducibility of microbial mutagenicity assays. 2. Testing of carcinogens and noncarcinogens in Salmonella typhimurium and E. coli. Environ Mutagen 1985;7 Suppl. 5: 1248 (cited in Stouten et al., 2000).

ECHA REACH registration dossier (2020). 1,2,3-benzotriazole. https://echa.europa.eu/registration-dossier/-/registered-dossier/14234/7/2/1 Accessed August 2020.

Health Council of the Netherlands: Dutch expert committee on occupational standards (DECOS). 1,2,3-Benzotriazole. The Hague: Health Council of the Netherlands, 2000; publication no. 2000/14OSH. Available at: file:///C:/Users/Admin/Downloads/advisory-report-1-2-3-benzotriazole-health-based-recommended-occupational-exposure-limit%20(1).pdf [accessed June 2020].

National Toxicology Program. Bioassay of 1H-benzotriazole for possible carcinogenicity. National Cancer Institute Carcinogenesis Technical Report Series. 1978; 88:1-131. Available at: https://pubmed.ncbi.nlm.nih.gov/12830211/ [accessed June 2020].

Stouten H, Rutten A, van de Gevel I, De Vrijer F. (2000) - The Nordic Expert Group for Criteria Documentation of Health Risks from Chemicals and The Dutch Expert Committee on Occupational Standards 126. 1,2,3-Benzotriazole. Available at: https://gupea.ub.gu.se/bitstream/2077/4230/1/ah2000_24.pdf [accessed June 2020].

Zeiger E, Anderson B, Haworth S, Lawlor T, Mortelmans K, Speck W. Salmonella mutagenicity tests. 3. Results from the testing of 255 chemicals. Environ Mutagen 1987;9, Suppl. 9:1-110 (cited in Stouten et al., 2000).

https://echa.europa.eu/registration-dossier/-/registered-dossier/14234/7/2/1

https://pubmed.ncbi.nlm.nih.gov/12830211/

https://gupea.ub.gu.se/bitstream/2077/4230/1/ah2000_24.pdf


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Section B: 1H-benzotriazole, 4(or 5)-

methyl-

Section B1.0 Chemical identification, use and


Section B1.1 Reasons for consideration

A report by WRc (2014) assessed the risk to drinking water from the use of 1H-benzotriazole, 4(or 5)-methyl- as a corrosion inhibitor in dishwasher detergents. In the UK, consumption decreased from 832 tonnes/year in 2007 to 390 tonnes/year in 2012 (WRc, 2014). Levels of 1H-benzotriazole, 4(or 5)-methyl- in drinking water were measured as 0.07 µg/L and estimated to be in the range 1.14 – 20.3 µg/L; the authors noted that this was based on estimated effluent concentrations 3.5 times higher than measured concentrations reported in the literature. 1H-benzotriazole, 4(or 5)-methyl- is included for further consideration here as the estimated exposures through drinking water and bathing were found to be greater than the estimated exposure (SED of 0.00002 mg/kg bw/day) through intended use at the maximum predicted levels (WRc, 2014).

Section B1.2 Identification and physicochemical properties.

1H-benzotriazole, 4(or 5)-methyl- is a heterocyclic compound. CAS no. 29385-43-1 (Tolyltriazole) identifies the commercial mixture composed of approximately equal amounts of 4- and 5-methylbenzotriazole, with small quantities of the 6- and 7- methyl isomers (TNO BIBRA, 1998 – cited in Beltoft et al., 2013). Identifiers and properties of 1H-benzotriazole, 4(or 5)-methyl- are given in Table B1.1.

Section B1.3 Hazard classifications

1H-benzotriazole, 4(or 5)-methyl- has been assigned a hazard category of Acute Tox 4, with hazard statement H302 – “harmful if swallowed” in the REACH dissemination page (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020).

The notified EU classification and labelling6 according to CLP regulation (EC) No. 1272/2008, is Acute Tox. 4 with hazard 302 – “harmful if swallowed”. There is no harmonised classification.

Section B1.4 Occurrence, production and use.

1H-benzotriazole, 4(or 5)-methyl- is produced by reaction of o-phenylenediamine with nitrous acid in the presence of glacial acetic acid or by reaction of hydrochloric acid or nitrous acid with o-phenylenediamine. It is used as an inhibitor of corrosion of copper and copper alloys, in antioxidants, and photographic developers (Beltoft et al., 2013). As detailed above, the annual production of 1H-benzotriazole, 4(or 5)-methyl- in the EU in 2012 was 390 tonnes.

6 https://echa.europa.eu/information-on-chemicals/cl-inventory-database/-/discli/details/15467

https://echa.europa.eu/information-on-chemicals/cl-inventory-database/-/discli/details/15467


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No information relating to the levels used in dishwasher products was identified in the public domain.

Table B1.1: 1H-benzotriazole, 4(or 5)-methyl- identification and physicochemical properties


Cas No

29385-43-1

IUPAC name 5-methyl-1H-1,2,3-benzotriazole

Chemical Group Heterocyclic

Physical form Light brown granules

DSL name Not on list

Common name Methyl-1H-benzotriazole

Formula: C7H7N3


Structure

Read across chemical – 1,2,3-benzotriazole

Solubility (water) <0.1 g/l (at 18°C).

Melting point 76-87°C

Boiling point 210-212°C (at 12 mm Hg), 160°C (at 2 mm Hg)

Density 1.24 g/mL

Log Pow 1.08 (at 25oC)

Vapour Press. 0.03 mmHg (Pa) (at 20°C)

Sources: WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl- 2020.

Section B1.5 Human exposure

Occupational exposure is the most likely source of exposure to 1H-benzotriazole, 4(or 5)-methyl- in humans, with dermal and inhalation routes being key. Non-occupational exposure may occur from the use of DCPs or through ingestion of drinking water that has been contaminated with 1H-benzotriazole, 4(or 5)-methyl- (WRc, 2014).


35

Section B2.0 Human relevant health effects

Section B2.1 Toxicokinetics

No data was identified relating to the toxicokinetics of 1H-benzotriazole, 4(or 5)-methyl- in humans or experimental species following exposure via any route.

In the absence of data, the REACH registration document utilises the physicochemical properties of 1H-benzotriazole, 4(or 5)-methyl- to estimate its toxicokinetics (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020). Absorption via the oral and dermal route were assumed to be 100 %, with inhalation exposure considered unlikely to occur. Bioaccumulation potential was also considered to be low, with minimal phase I metabolism occurring. Elimination was predicted to occur through urine and faeces (Beltoft et al., 2013).

Section B2.2 Acute toxicity

No data were identified relating to the acute effects of 1H-benzotriazole, 4(or 5)-methyl- in humans via any route of exposure.

The acute oral toxicity of 1H-benzotriazole, 4(or 5)-methyl- has been tested in rats and mice. Acute oral toxicity studies (carried out prior to OEC/GLP standards) have reported LD50 values in rats between 675 and 3400 mg/kg bw. In mice, an oral LD50 value of 800 mg/kg bw has been reported (TNO BIBRA 1998 – cited in Beltoft et al., 2013).

In rats, 500 mg/kg bw and above of tolytriazole given by oral gavage caused central nervous system effects. Effects on lungs, stomach and liver were seen at doses of 1000 mg/kg bw and above. Death occurred at higher doses, generally 2000 mg/kg bw and above (no further details given; it is not clear whether the test material was tolyltriazole or 5-methylbenzotriazole) (TNO BIBRA 1998 – cited in Beltoft et al., 2013).

In a study carried out to OECD Guideline 401 (Acute Oral Toxicity), sublethal effects were noted in rats administered 1H-benzotriazole, 4(or 5)-methyl- by oral gavage at doses of between 500 and 2000 mg/kg bw). Non-lethal clinical signs of toxicity included central nervous system effects, and an LD50 of 720 mg/kg bw was determined (TNO BIBRA 1998 – cited in Beltoft et al., 2013).

Based on the acute lethal toxicity data and using EC-classification criteria, 1H-benzotriazole, 4(or 5)-methyl- is considered to be harmful following oral exposure (Beltoft et al., 2013).

Section B2.3 Repeat dose toxicity

Section B2.3.1 Systemic effects

No data were identified relating to the systemic effects of 1H-benzotriazole, 4(or 5)-methyl- in humans via any route of exposure. The REACH registration document reports the effects of short-term dietary exposure of Wistar rats to 1H-benzotriazole, 4(or 5)-methyl- , in an unnamed study (1998) carried out to


36

OECD Guideline 407 (Repeated Dose 28-day Oral Toxicity in Rodents)(ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020). Rats were exposed to 1H-benzotriazole, 4(or 5)-methyl- at doses of 0, 50, 150 and 450 mg/kg bw/day via the diet. No adverse effects were apparent at doses ≤150 mg/kg bw/day. At the highest dose, toxicity was evident as apathy following gavage, reduced levels of erythrocytes, haematocrit and haemoglobin in males, raised activity of alanine-aminotransferase, and reduced plasma protein concentration in both males and females.

Section B2.3.2 Irritancy and corrosivity

No data were identified relating to the irritancy potential of 1H-benzotriazole, 4(or 5)-methyl- in humans (see also section 2.3.3).

In a study carried out to OECD Guideline 404 (Acute Dermal Irritation/Corrosion) 1H-benzotriazole, 4(or 5)-methyl- did not cause skin irritation in the rabbit at a dose of 500 mg when applied under semi-occlusive conditions for 4 hr to intact clipped skin (observation up to 14 days post exposure), and therefore does not meet the CLP requirements for classification as a skin irritant (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020).

An acute eye irritation study carried out to OECD Guideline 405 (Acute Eye Irritation/Corrosion) was reported in the rabbit in which 1H-benzotriazole, 4(or 5)-methyl- resulted in mild irritation. As such, it did not meet the criteria for classification under CLP regulations (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020).

Section B2.3.3 Sensitisation

Data relating to the sensitising potential of 1H-benzotriazole, 4(or 5)-methyl- in humans could not be identified.

1H-benzotriazole, 4(or 5)-methyl- was reported to be non-sensitising in a guinea pig maximisation test carried out to OECD Guideline 406 (Skin Sensitisation) (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020).

Section B2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of 1H-benzotriazole, 4(or 5)-methyl- in humans by any route of exposure.

The Danish EPA report that 1H-benzotriazole, 4(or 5)-methyl- was mutagenic or weakly mutagenic to Salmonella typhimurium strains in the presence but not in the absence of S9 (no further details given) (Beltoft et al., 2013).

1H-benzotriazole, 4(or 5)-methyl- has been tested in the HPRT forward mutation assay in CHO cells according to OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test) and was negative in the absence of metabolic activation at a dose of 600 mg/kg bw (Beltoft et al., 2013).

The Danish EPA also state that 1H-benzotriazole, 4(or 5)-methyl- did not cause the transformation of mouse cells and did not damage the DNA of human lung cells. However, no


37

metabolic activation systems were used in these studies (no further details given) (Beltoft et al., 2013).

The cytogenic potential of 1H-benzotriazole, 4(or 5)-methyl- has been assessed in an in vivo study carried out to OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test). A single oral gavage dose of 600 mg/kg/bw (previously shown to cause toxicity) administered to male and female mice was not associated with an increase in the incidence of micronucleated polychromatic erythrocytes at either 24, 48 or 72 h post exposure (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020).

It should be noted that the REACH Registration document does not include consideration of the in vitro study findings for mutagenicity and concludes that there is no evidence for genotoxicity (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020). Overall, considering the weight of evidence from the genotoxicity data package, the greatest weight can be given to the in vivo micronucleus test (OECD guideline 474) which was favourable. In vitro data from a number of types of study are supportive of 1H-benzotriazole, 4(or 5)-methyl- being non-mutagenic.

Section B2.3.5 Carcinogenicity

No data relating to the carcinogenic potential of 1H-benzotriazole, 4(or 5)-methyl- in humans or experimental species were identified.

In the absence of carcinogenicity data on 1H-benzotriazole, 4(or 5)-methyl-, 1,2,3-benzotriazole could be considered for use as a surrogate chemical for a read across approach. However, the mutagenicity and carcinogenicity databases are also considered inadequate to allow classification of 1,2,3-benzotriazole fully with regard to its carcinogenic potential.

QSAR modelling was carried out (by the authors of this report) for 1H-benzotriazole, 4(or 5)-methyl- using ToxTree, VEGA and the OECD Toolbox.

In ToxTree, 1H-benzotriazole, 4(or 5)-methyl- was negative for genotoxic and non-genotoxic carcinogenicity.

The reliability of the VEGA model was compromised by the presence of a fragment that has never been found in the model's training set.

In the OECD Toolbox, there were no alerts for DNA binding or DNA alerts for Ames, chromosomal aberrations or micronuclei.

Section B2.3.6 Reproductive and developmental toxicity

No data relating to the reproductive and/or developmental toxicity of 1H-benzotriazole, 4(or 5)-methyl- in humans or experimental species was identified via any route of exposure.

In the absence of data, the REACH registration document uses read-across from 1,2,3-benzotriazole, which is considered sufficiently similar, meeting criteria set out in Annex XI, 1.5, to allow the approach for this endpoint (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020). The document reports an unnamed reproductive toxicity study carried out to OECD Guideline 421 (Reproduction / Developmental Toxicity


38

Screening Test) in which 1,2,3-benzotriazole was administered by oral gavage to male and female Wistar rats at doses of 0, 12.5, 50 and 200 mg/kg bw/day. No clinical or general signs of toxicity were apparent and there was no indication that reproductive performance had been adversely affected in either sex. Histopathology of the ovaries and epididymides/testes was also unremarkable. No developmental toxicity was evident in the offspring of exposed parents. It is noted by the registrant that OECD Guideline 421 does not provide complete information concerning all aspects of reproduction and development or post-natal effects that may become apparent due to pre-natal exposure (ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020).

Section B2.3.7 Specific considerations

No further toxicity studies were identified.

Section B2.3.8 Summary of human relevant health effects.

• No data relating to the potential acute toxicity of 1H-benzotriazole, 4(or 5)-methyl- in humans were identified.

• Toxicokinetic data is absent for both humans and experimental species. Bioaccumulation potential has been estimated as low and excretion is considered to occur via urine and faeces.

• According to EC-classification criteria, 1H-benzotriazole, 4(or 5)-methyl-is considered harmful via the oral route.

• Based on the available data 1H-benzotriazole, 4(or 5)-methyl- does not have irritation or sensitising potential.

• The overall weight of evidence suggests 1H-benzotriazole, 4(or 5)-methyl- is non-mutagenic, which is based on OECD guideline an in vivo micronucleus assay.

• There are no carcinogenicity data for 1H-benzotriazole, 4(or 5)-methyl- and read across using 1,2,3-benzotriazole is also not possible due to the poor data set available and lack of a read across justification. QSAR models suggest 1H-benzotriazole, 4(or 5)-methyl- to be non-carcinogenic but confidence in the ability to use these models for a tautomeric chemical is low.

• Repeat dose (28-day) oral studies showed adverse systemic effects of 1H-benzotriazole, 4(or 5)-methyl- at doses >150 mg/kg bw/day.

• A reproductive and developmental toxicity study did not show adverse effects following oral exposure.

Section B3.0 Identification of relevant point(s) of


Section B3.1 Critical endpoint for human health risk assessment

purposes

The POD for use in the risk assessment (see Section B4.0) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to 1H-benzotriazole, 4(or 5)-methyl through any exposure route. However, a number of oral repeated dose


39

experimental studies have been carried out for 1H-benzotriazole, 4(or 5)-methyl-, for systemic and specific organ toxicity. These are summarised in Table B3.1.


40

Table B3.1 Oral route toxicity studies for 1H-benzotriazole, 4(or 5)-methyl-



OECD 407 Repeated dose/28 days

Rat / Wistar 50 – 450 via the diet At the highest dose – toxicity evident as apathy following gavage, reduced levels of erythrocytes, haematocrit and haemoglobin in males, raised activity of alanine-aminotransferase and reduced plasma protein concentration in males and females. NOAEL of 150 mg/kg bw/day can be identified based on systemic toxicity.

ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020 (unnamed study 1998)

OECD 421 Reproduction / Developmental Toxicity Screening Test

Rat / Wistar READ-ACROSS using 1,2,3-benzotriazole 12.5 – 200 via oral gavage

No treatment related effects on reproductive performance in either sex at any dose. No developmental toxicity apparent at any dose. A NOAEL of 200 mg/kg bw/day (highest dose tested) was derived for reproductive and developmental toxicity.

ECHA REACH registration dossier for 1H-benzotriazole, 4(or 5)-methyl-, 2020 (unnamed study 2012)

Shaded row represents the study considered suitable for use in the risk assessment


41

The data indicate that the systemic effects are the most sensitive following oral exposure to 1H-benzotriazole, 4(or 5)-methyl- (discussed further in section B3.3). There is no carcinogenicity data available for this chemical and read-across using 1,2,3-benzotriazole is not possible, also due to limited data. There is no evidence from currently available studies that 1H-benzotriazole, 4(or 5)-methyl- causes reproductive or developmental toxicity.

Section B3.2 Current health-based guidance values


Section B3.3 Identification of POD to be used for the risk

assessment.

The lowest effect level following repeated oral exposure to 1H-benzotriazole, 4(or 5)-methyl- was reported in a 28-day study and was associated with systemic toxicity, resulting in a NOAEL of 150 mg/kg bw/day.

Section B4.0 Drinking water risk assessment


Few adverse effects were seen following oral exposure to 1H-benzotriazole, 4(or 5)-methyl-.


Based on the data obtained, the NOAEL is considered to be 150 mg/kg bw/day based on haematological effects.


An AF of 600 is considered appropriate.

POD (mg/kg bw/day) 150

AF Justification

Interspecies differences 10 POD based on animal data

Intraspecies differences 10 To account of human variability

Use of subchronic data 3 NOAEL based on a 28 day study

Inadequate databases 2 No carcinogenicity data available.

Total AF 600


Based on a NOAEL of 150 mg/kg bw/day and an AF of 600, the proposed TDI is 250 µg/kg bw/day.




• 500 µg/L for a 10 kg child drinking 1 L water per day

• 333 µg/L for a 5 kg infant drinking 0.75 L water per day


42

Exposure assessment

The concentration of 1H-benzotriazole, 4(or 5)-methyl- measured in drinking water, as reported by WRc (2014), was 0.07 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.


• 0.002 µg /kg bw/day for a 60kg adult drinking 2L a day

• 0.007 µg /kg bw/day for a 10kg child drinking 1L a day

• 0.105 µg /kg bw/day for an 5kg infant drinking 0.75L a day This assumes that total exposure is due to exposure via drinking water.


The maximum intake of 1H-benzotriazole, 4(or 5)-methyl- via drinking water by adults, children and infants is less than the TDI (HQ <1) and the measured concentration of 1H-benzotriazole, 4(or 5)-methyl- in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to 1H-benzotriazole, 4(or 5)-methyl- via drinking water.

Risk communication

The MOEs for 1H-benzotriazole, 4(or 5)-methyl, based on the NOAEL of 150 mg/kg bw/day and measured intakes of 0.002, 0.007 and 0.105 µg /kg bw/day, are 64285714, 21428571, and 14285714 for adults, children and infants respectively. As MOEs are > 100 this indicates that exposures are not of concern in terms of risk to public health.

References

Beltoft V., Nielsen E., Ladefoged O. (2013) - The Danish Environmental Protection Agency. 1526 Benzotriazole and Tolyltriazole. Evaluation of health hazards and proposal of health based quality criteria for soil and drinking water. Available at: https://www2.mst.dk/Udgiv/publications/2013/12/978-87-93026-81-0.pdf [accessed June 2020].

ECHA REACH registration dossier (2020). 1H-benzotriazole, 4(or 5)-methyl-. https://echa.europa.eu/registration-dossier/-/registered-dossier/14272/7/2/2 [accessed August 2020].

TNO BIBRA (1998). Toxicity Profile, Tolyltriazole. 3rd ed. TNO BIBRA International Ltd. Great Britain – cited in Beltoft et al., 2013.

https://www2.mst.dk/Udgiv/publications/2013/12/978-87-93026-81-0.pdf

https://echa.europa.eu/registration-dossier/-/registered-dossier/14272/7/2/2


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Section C: (1-hydroxyethylidene)

diphosphonic acid (HEDP)

Section C1.0 Chemical identification, use and


Section C1.1 Reasons for consideration

The WRc (2014) report assessed the use of HEDP as a phosphonate in dish, laundry and household cleaning products. UK consumption of these products decreased from 2310 tonnes/year in 2007 to 2040 tonnes/year in 2012. Levels of HEDP in drinking water were estimated to be in the range 10.3-20.5 µg/L. HEDP is included for further consideration here as the estimated exposures through drinking water and bathing were greater than the estimated exposure (SED of 0.0005 mg/kg bw/day) through intended use (WRc 2014).

Section C1.2 Identification and physicochemical properties.

Being multifunctional acids, phosphonates will form salts or complexes of different composition, depending on the chemical composition and the pH of the environment (HERA, 2004). These phosphonates are primarily used as acids and sodium salts. Their behaviour in the body does not depend on the presence of sodium as the counter ion. Therefore, phosphonate salts may be used to assess the toxicity of the phosphonate acid, although it should be noted that the toxicity of the salts in some cases i.e. for HEDP is greater than for the acid. This is considered to be related to the perturbation of iron homeostasis as a result of the chelating properties of HEDP. In this case, only data for HEDP was used to assess repeat dose toxicity. However, Hera (2004) stated ‘There is, however, no need to distinguish between the acid and salt form of the respective phosphonic acid. The basic justification for this is that in dilute aqueous conditions of defined pH a salt will behave no differently to parent acid. The effect of the counter-ion (usually sodium) is not judged to be significant’.

Table C1.1 HEDP identification and physicochemical properties


Cas No

2809-21-4

IUPAC name (1-hydroxy-1-phosphonoethyl)phosphonic acid

Chemical Group Phosphonate

Physical form Liquid

DSL name Phosphonic acid, (1-hydroxyethylidene)bis-

Common name Etidronic acid

Formula C2H8O7P2



44


Structure

Solubility Very soluble in water

Melting point 195 °C

Boiling point 456.79 °C

Density 1.45 g/cm3 (20 °C)

Log Pow -3.49 (25 °C)

Vapour Press. 0 Pa (25 °C)

Sources: WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for Etidronic acid 2020)

Section C1.3 Hazard classifications

HEDP has been assigned a hazard category of Acute Tox. 4, with hazard statement H302 – “harmful if swallowed” and Eye Dam. 1 with hazard statement H318 – “causes serious eye damage” in the REACH dissemination page (ECHA REACH registration dossier for Etidronic acid 2020).

The EU notified classification and labelling according to CLP regulation (EC) No. 1272/2008 is Eye Dam. 1 with hazard statement H318 – causes serious eye damage, Acute Tox. 4 with hazard statement H302 – harmful if swallowed, Skin Corr. 1A, 1B and 1C with hazard statement H314 – causes severe skin burns, Skin Irrit. 2 with hazard statement H315 – causes skin irritation and STOT RE3 with hazard statement H335 – may cause respiratory irritation. There is no harmonised classification.

Section C1.4 Occurrence, production and use.

Phosphonates, including HEDP, are used in a wide range of products, including regular and compact laundry detergents, fabric conditioners, laundry additives, hand and machine dishwashing detergent and surface, carpet and toilet cleaners (HERA 2004).

Section C1.5 Human exposure

Exposure to HEDP is predominantly during pre-treatment of laundry as well as direct contract from hand washing laundry, hand dishwashing and using hard surface cleaners, indirect contact from wearing laundered clothes, inhalation of laundry powder dust and aerosol particles, and oral exposure of residues on eating utensils and dishes as well as from exposure to residues found in water (HERA 2004).

Section C2.0 Human relevant health effects

Section C2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of HEDP in humans.


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In a dog study, three female Beagle dogs were administered a single dose of 20 mg/kg bw 32P-HEDP or 50 mg/kg bw 14C-HEDP by gavage. Over 72 hours, 21 % was absorbed in young dogs and 14 % in older dogs, primarily from the stomach. 80 % of the radioactivity recovered was found in faeces and 10 % in urine (unknown author cited in HERA 2004).

Monkkonen et al. investigated the distribution of i.v. administered 14C-labeled HEDP in MRI mice. Forty male mice were given a single i.v. injection of 25 mg/kg bw 14C-HEDP. Organ and blood samples were collected until 360 days after treatment. Radioactivity was found to be removed rapidly from the plasma, transferred to kidney and tibia/femur with trace levels present in spleen and liver. Levels in kidneys decreased rapidly, consistent with rapid urine excretion. Small amounts of 14C remained in the long bones 360 days after treatment (Monkkonen et al., 1987, cited in HERA 2004).

The distribution of 14C-labeled HEDP acid was also studied in Sprague-Dawley rats for up to 16 days after intraperitoneal (i.p.) injection (Larsson and Rohlin, 1980, cited in HERA 2004). One- and 5-day old rats (sex not stated) were given a single i.p. injection of 50 mg/kg bw 14C-HEDP. As seen with i.v. injection, a time dependent increase in 14C was seen in peripheral bone surfaces and epiphyseal cartilage of long bones and trace amounts in kidneys, liver, stomach and renal medulla.

Section C2.2 Acute toxicity

The acute toxicity of HEDP has been tested in several experimental species. In a study carried out according to OECD 401 (Acute Oral Toxicity), an LD50 of 1878 mg/kg was determined. Clinical signs of toxicity included weakness within the first few minutes, dyspnoea and collapse. At necropsy, inflammation of the gastric mucosa and haemorrhagic areas in the lungs were observed (Younger Laboratories, 1965 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Etidronic acid 2020; Monsanto unpublished report cited in IUCLID 2000).

In a similar study, an LD50 of 1440 mg/kg bw was determined. Decreased appetite and activity, increasing weakness, diarrhoea, tremors, collapse and death were all observed. (Younger Laboratories, 1977 cited in HERA 2004; ECHA REACH registration dossier for Etidronic acid 2020). Although both these studies were pre-GLP and not in full compliance with OECD guidelines, they appear to be well conducted and were therefore judged to be reliable (HERA 2004).

Other studies have reported LD50 values of between 1008 and >5000 mg/kg bw in rats and between 1100 and 4120 mg/kg bw in mice (ECHA REACH registration dossier 2020 for Etidronic acid; HERA 2004; IUCLID 2000; OECD SIDS 2004; CIR 2016). A number of acute dermal test in rabbits were identified. LD50 values of 7940 – 10000 mg/kg have been reported. Clinical signs include weakness, reduced appetite and activity (CIR 2016; HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004).

Section C2.3 Repeat dose toxicity

Section C2.3.1 Systemic effects

A number of repeated dose toxicity studies were found.


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A 28-day study was carried out in which male and female Wistar rats (number not given) were administered 0, 0.3, 3, 30 or 100 mg/kg bw/day HEDP via gavage twice daily for 28 days. No adverse effects were reported and a NOAEL of 100 mg/kg bw/day was determined (highest dose tested) (Monsanto, date unknown cited in IUCLID 2000).

The Cosmetic Ingredient Review (CIR, 2016) in the USA cited three pre-2013 short term in vivo studies. One was a 28-day oral gavage study in rats from which a NOAEL of 30 mg/kg bw/day was determined (no further details available) (CIR, 2016; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004). The other two studies, a 28-day study in rats (Peter et al., 1998 cited in CIR, 2016) and a study in cats (duration not given; Jowsey et al., 1970 cited in CIR, 2016) were carried out with a HEDP salt and did not determine a point of departure as insufficient data were presented.

A number of 90-day studies were carried out. In the first study, carried out according to OECD 408 (Repeated dose 90-day oral toxicity study in rodents), Charles River albino rats (15/sex/dose) were administered 0, 1000, 3000 and 10000 ppm HEDP (0, 154, 524 and 1583 mg/kg bw/day for males and 0, 166, 545 and 1724 mg/kg bw/day for females) via the diet for 90 days (Industrial Biotest Labs Inc., 1979 cited in HERA 2004; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004). Body weights and absolute and relative liver weights were slightly decreased in the highest dose groups. An increased erythrocyte count, reduced haemoglobin concentration and decreased haematocrit was reported in males only. No treatment-related effects on clinical chemistry, urine analysis or gross pathology were observed. The authors stated that there were no treatment-related histopathological effects. However, in the HERA document, it was noted that bilateral mineralized micro-concretions in kidney tubules and extramedullary haematopoiesis in the spleen was observed in the high dose groups and not in the controls. The extramedullary haematopoiesis was probably related to alterations in iron homeostasis, and bilateral mineralized micro-concretions may be a result of altered calcium homeostasis. A NOAEL of 1583 mg/kg bw/day was determined (highest dose tested).

In a similar study, Charles River albino rats (15 per dose group) were administered 0, 3000, 10000 or 30000 ppm HEDP (180, 600 or 1800 mg/kg bw/day) for 90 days via the diet according to OECD 408 (Repeated dose 90-day oral toxicity study in rodents). A NOAEL of 600 mg/kg bw/day was determined based on decreased food intake and body weight gain, increased red blood cells in males, and decreases in white blood cells, haemoglobin and haematocrit and liver weight in females at 1800 mg/kg bw/day (Monsanto, 1981 cited in IUCLID 2000; ECHA REACH registration dossier for Etidronic acid 2020).

Two studies were also carried out in dogs. In the first study, Beagle dogs (number not given) were administered 0, 25, 75 or 250 mg/kg bw/day via the diet seven days per week for 90 days. A NOAEL of 250 mg/kg bw/day was determined (highest dose tested) (Monsanto, 1981 cited in IUCLID 2000).

The second study in dogs was carried out according to OECD 409 (Repeated dose 90-day oral toxicity study in non-rodents). Beagle dogs (4/sex/group) were administered 0, 1000, 3000 and 10000 ppm HEDP (0, 191, 554 and 1746 mg/kg bw/day for males and 0, 202, 553 and 1620 mg/kg bw/day for females) via the diet for 90 days (Industrial Biotest Labs Inc., 1975 cited in HERA 2004; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS


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2004). Erythrocyte counts were slightly elevated in all test groups except the low dose male group, mean corpuscular volume was slightly decreased at the two highest dose levels, and a dose-related increase in leukocytes in the urine was seen at all doses. However, no histopathological changes were observed in the haematopoietic or urinary systems and therefore these observations were not considered to be of toxicological concern. A NOAEL of 1620 mg/kg bw/day was determined (highest dose tested).

The disodium salt of HEDP was also tested in two 90-day studies in rats, as well as in a 2-year study. In the first 90-day study (Huntingdon Research Centre, 1977 cited in HERA 2004), Sprague-Dawley rats (10/sex/dose) were administered 0, 500, 2000 and 10000 ppm HEDP disodium salt in the diet (0, 41, 169 and 817 mg active salt/kg bw/day for males, and 0, 50, 195 and 1000 mg active salt/kg bw/day for females). Severe pallor of skin in rats from the high dose group and slight pallor in rats in the mid dose group were observed. A decrease in red cell parameters was seen in the highest dose group for both sexes, and for males at 2000 ppm as well as prolonged anaemia in both sexes at 10000 ppm, with a slight retardation of bone marrow development. The observed anaemia, reduction in red cell parameters and pallor are all consistent with the perturbation of iron homeostasis. Some of the effects were also observed at 2000 ppm and therefore a NOAEL of 41 mg active salt/kg bw/day was determined.

In the second 90-day study, Charles River rats (20/sex/dose) were fed 200 and 1000 ppm HEDP disodium salt (260 and 1300 mg/kg bw/day)(Nixon et al., 1972 cited in HERA 2004). The only effect noted was a slight increase in relative kidney weight in females treated with 1300 mg/kg bw/day, which occurred in the absence of histopathological findings. A NOAEL of 260 mg/kg bw/day was determined (highest dose tested). The study was non-GLP and few details were available.

The 2-year oral feeding study in rats was carried out according to OECD 452 (Huntingdon Research Centre, 1979 cited in HERA 2004). Sprague-Dawley rats (40 sex/dose) were administered the test substance via the diet at dose levels of 0, 500, 2000 and 10000 ppm (0, 19, 78, 384 mg active salt/kg bw/day for males and 0, 24, 96, 493 mg active salt/kg bw/day for females) for 104 weeks. No increased incidence of neoplastic lesions was observed in the treated groups at study termination, and no mortality or treatment-related effects were observed on clinical chemistry or gross pathology. Several haematological perturbations were seen during the study, but there were no treatment-related effects which persisted until the termination at 24 months. Anaemia was observed in animals in the mid and high dose groups. A lack of iron in the spleen was seen in the two highest dose levels at 26 weeks, although this resolved by 104 weeks. The iron deficiency was probably related to the chelating properties of the test substance. A NOAEL 19 mg active salt/kg bw/day based on anaemia at 78 mg/kg bw/day. The study was of good quality.

Section C2.3.2 Irritancy and corrosivity

No data were identified relating to the irritancy potential of HEDP in humans.

A number of skin and eye irritation studies have been carried out in animals. 0.5 mL HEDP (aqueous solution containing 60 % HEDP and 0.02 % HCl) was applied to the skin of six New Zealand white rabbits under an occlusive dressing for 24 hours (Younger Laboratories, 1977 cited in HERA, 2004; Monsanto, 1977 cited in IUCLID 2000; OECD SIDS 2004). No irritation was


48

observed throughout the study. However, this study was not conducted in compliance with OECD guidelines and GLP regulations.

Another skin irritation study was carried out in which an aqueous solution (concentration not stated) was administered to the intact and abraded skin of three New Zealand white rabbits under occlusive dressing for 24 hours (Younger Laboratories, 1965a cited in HERA, 2004; Monsanto, 1965 cited in IUCLID 2000; OECD SIDS 2004). Moderate skin irritation was observed, although it is unknown if irritation was seen on intact or abraded skin as results were not shown separately. Moreover, there was no distinction between erythema/eschar and oedema formation; hence this non GLP/OECD study was judged unreliable (HERA 2004).

In an eye irritation study, 0.1 mL HEDP (aqueous solution containing 60 % HEDP and 0.02 % HCl) was placed into the conjunctival sac of the right eye of three New Zealand white rabbits (Younger Laboratories, 1965a cited in HERA 2004; Monsanto, 1965 cited in IUCLID 2000; OECD SIDS 2004). Eyes were rinsed after four seconds (one animal) or 24 hours (2 animals) and the animals were evaluated up to 7 days after dosing. Moderate lachrymation, mild oedema and erythema and mild corneal cloudiness were observed after 1 hour (overall score 31) in the animal whose eye was rinsed four seconds after treatment. These effects were reduced to very slight redness after 7 days. In animals whose eyes were not immediately rinsed, copious discharge, translucent cornea with iris details moderately obscured and swelling with partial eversion of lids was seen after one hour, which increased in severity throughout the observation period. On day 7, the iris did not respond to light and the lower half of the cornea was opaque. Authors of the HERA document concluded that HEDP was moderately to severely irritating, whereas OECD SIDS considered it to be corrosive. Although predating GLP and OECD guidelines, the study was conducted using an acceptable protocol and provided reliable information on the eye irritation potential of HEDP (HERA 2004).

In a second study, 0.1 mL HEDP (aqueous solution containing 60 % HEDP and 0.02 % HCl) was instilled into the eyes of six rabbits for 24 hours, and animals were observed for up to 21 days (Younger Laboratories, 1977 cited in HERA 2004; Monsanto, 1977 cited in IUCLID 2000; OECD SIDS 2004). Initial pain, severe erythema and copious discharge were observed 10 minutes after administration which became more pronounced after 24 hours but then improved slightly over the next 10 days. However, after 14 days ulceration was observed in all rabbits which was not reversible within the 21 days observation period. One animal also had a ruptured cornea. At 21 days, corneal ulceration, slight erythema and copious discharge were still observed in 4 animals. The authors concluded that HEDP is moderately to severely irritating to rabbit’s eyes, and effects are non-reversible.

A number of other studies have been carried out in rabbits in which HEDP was slightly irritating, irritating or corrosive to the skin and/or eye (IUCLID 2000; OECD SIDS 2004). Few details are available.

Section C2.3.3 Sensitisation

No data were identified relating to the sensitisation of HEDP in humans or animals.

In the absence of data for HEDP, HEDP sodium salt has been used as a surrogate chemical in a read across approach.


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A disodium salt of HEDP was negative in a guinea pig maximisation test (Henkel, 1983 cited in HERA 2004). In the induction phase, 20 animals were injected intracutaneously with 0.1 mL Freund’s Adjuvant/water, 5 % aqueous solution of HEDP salt or 5 % aqueous solution of 1:1 mixture HEDP salt and Freund’s Adjunct in the shoulder region. Twenty animals were used as controls, receiving also intracutaneous injections of Freund’s Adjuvant, water or 1:1 mixture of 5 % water and Freund’s Adjunct. After a week, the treatment group were treated with a mixture containing 5 % HEDP salt in Vaseline was placed on the injection site under occluded conditions for 48 hours. Two weeks after the induction phase, the flanks of the treated and the control animals were shaved and a challenge patch containing 25 % HEDP salt in Vaseline was applied to one flank of the animals for 21 hours. No occlusive patch appeared to have been used. Skin reactions was assessed approximately 24 hours from the start of the challenge application. HEDP acid did not cause skin sensitisation in guinea pigs. However, the the study was not in compliance with GLP nor OECD standards but was reported in enough detail to judge it as scientifically reliable (HERA 2004).

QSAR modelling

Due to the absence of experimental data for HEDP, quantitative structure-activity relationship (QSAR) modelling has also been carried out (by the authors of this report) to identify the presence of structural alerts for sensitisation. Predictions were made for sensitisation using Toxtree Version 3.1.0-1851-1525442531402, VEGA Version 1.1.5-b22 (Calculation core version: 1.2.8) and OECD Toolbox Version 4.4.

In ToxTree, no skin sensitisation reactivity domains alerts were identified.

In VEGA, using the CAESAR and IRFMN/JRC models, HEDP was predicted to be a sensitiser. However, this prediction should be treated with caution. Both models may not be reliable as HEDP is outside the Applicability Domain of both models. There were also other issues with the prediction that make it unreliable. These include only moderately similar compounds with known experimental value being found in the training set, similar molecules found in the training set having experimental values that disagree with the predicted value, and a prominent number of atom centred fragments of the compound not being found in the compounds of the training set, or are rare fragments.

In the OECD Toolbox, there were no alerts for sensitisation or protein binding, and protein binding potency for both cystine and lysine in the Direct Peptide Reactivity Assay (DPRA) assay was <9 %, indicating no protein binding alert. The Toolbox also predicted that Globally Harmonised System (GHS) criteria were not met for sensitisation.

Section C2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of HEDP in humans.

An Ames test was carried out with Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538, with and without metabolic activation, in which an aqueous solution of HEDP (2700 µg/plate; limit dose due to cytotoxicity and solubility) was tested (Monsanto, 1977 cited in HERA, 2004; Monsanto, 1977 cited in IUCLID; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004). HEDP was negative for mutagenicity as no increase in revertants was observed.


50

A later study was carried out according to OECD 471 (Bacterial Reverse Mutation Assay) using TA102, with and without metabolic activation. HEDP was non-mutagenic in the assay. The REACH registration dossier states that this test should be considered in combination with the earlier Ames test, carried out to provide information on the missed strain (TA102) (Unnamed study, 2015 cited in ECHA REACH registration dossier for Etidronic acid 2020).

HEDP was also negative in an in vitro mammalian cell micronucleus test (OECD 487) carried out in human lymphocytes, with and without metabolic activation (unnamed study, 2015 cited in ECHA REACH registration dossier for Etidronic acid 2020).

HEDP (aqueous solution) was also evaluated for induction of mutations at the thymidine locus in a mouse lymphoma L5178Y assay (Litton Bionetics, 1978 cited in HERA 2004; Monsanto 1977 cited in IUCLID). The study was based on OECD method 476 (In Vitro Mammalian Cell Gene Mutation Test) but bromodeoxyuridine, a non-standard selective agent, was used. Two tests were carried out, with and without metabolic activation. In the first test, a dose-related increase in mutations with metabolic activation was observed, but as the relative growth was < 10 % the data were considered unreliable. In the second test, no dose-related responses were observed. Therefore, HEDP was not considered to be mutagenic. Although the study complied with OECD test guidelines, it was deemed to be of low reliability due to the high variability of control values, lack of details on test conditions, and the use of a non-standard selective agent, bromodeoxyuridine (HERA 2004).

Section C2.3.5 Carcinogenicity

No data were identified relating to the carcinogenic effects of HEDP in humans or animals.

In the absence of data for HEDP, HEDP sodium salt has been used as a surrogate chemical in a read across approach (see justification for read-across chemicals (Section C1.2)).

Disodium salt of HEDP was negative in a 2-year oral feeding study in rats, carried out according to OECD 452 (Huntingdon Research Centre, 1979 cited in HERA, 2004; ECHA REACH registration dossier for Etidronic acid 2020). Sprague-Dawley rats (40 sex/dose) were administered the test substance via the diet at dose levels of 0, 500, 2000 and 10000 ppm (0, 19, 78, 384 mg active salt/kg bw/day for males and 0, 24, 96, 493 mg active salt/kg bw/day for females) for 104 weeks. No mortality or treatment-related effects were observed on clinical chemistry or gross pathology and no treatment-related increase in neoplastic lesions were observed. The study was of good quality.

QSAR modelling

Due to the absence of experimental data with HEDP, QSAR modelling was also carried out (by the authors of this report) using ToxTree, VEGA and the OECD Toolbox to identify the presence of structural alerts for carcinogenicity.

In ToxTree, HEDP was negative for genotoxic and non-genotoxic carcinogenicity.

In VEGA, using the CAESAR and IRFMN models, HEDP was predicted to be carcinogenic but in the ISS, IRFMN/Antares and IRFMN/ISSCAN-CGX models it was predicted to be a non-carcinogen. However, all these predictions should be treated with caution. All models may


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not be reliable as HEDP is outside the Applicability Domain of all models. There were also other issues with the prediction that makes it unreliable. These include only moderately similar compounds with known experimental value being found in the training set, accuracy of prediction for similar molecules found in the training set not being optimal, similar molecules found in the training set having experimental values that disagree with the predicted value, and a prominent number of atom centred fragments of the compound not being found in the compounds of the training set, or are rare fragments.


Section C2.3.6 Reproductive and developmental toxicity

No data were identified relating to the reproductive and/or developmental toxicity of HEDP in humans.

Few specific reproductive or developmental studies were identified with HEDP.

In the 90-day study in rats with HEDP, described in Section 2.3.1, no treatment-related effects were observed in gonads, seminal vesicles, uterus or prostate. A NOAEL of 1583 mg/kg bw/day was determined (highest dose tested) (Industrial Biotest Labs Inc., 1979 cited in HERA 2004).

In the 90-day study in Beagle dogs treated with HEDP, described in Section 2.3.1, no treatment related effects on the reproductive organs were observed and a NOAEL of >1620 mg/kg bw/day was determined (highest dose tested) (Industrial Biotest Labs Inc., 1975 cited in HERA 2004). Both studies were pre-GLP but in compliance with OECD guideline 409. The quality of the studies is of unknown reliability (HERA 2004).

In a 2-generation study with Charles River rats (22/sex/dose for F0 and F1 and 20/sex/dose for F1b and F2), 0, 0.1 and 0.5 % disodium salt of HEDP (0, 112 or 447 mg active salt/kg bw/day) was administered via the diet to both sexes, or to pregnant females between gestation days 5-15 (Nolen and Buehler, 1971 cited in HERA 2004). The F0 females were allowed to deliver two litters (i.e., F1a, F1b) while a third was used for a teratology evaluation. Litters of F1b were used for breeding the F2 generation. No treatment-related effects on pregnancy rate were observed. The only effect seen was a significant decrease in the number of live pups born to dams in the top dose group, and a non-significant increase in stillborns, suggesting fetotoxicity. A reproductive NOAEL of 447 mg/kg bw/day (highest dose tested) and a developmental NOAEL of 112 mg active salt/kg bw/day based on fetotoxicity at the high doses. There was only limited information available with regard to the study design but deviations from the OECD guideline protocol were observed. These deficiencies limit the reliability of this investigation (HERA 2004).

In a similar study, pregnant rabbits were fed 0, 25, 50 or 100 mg active salt/kg bw/day HEDP disodium salt in the diet during day 2 to 16 of gestation (Nolen and Buehler, 1971 cited in HERA 2004). The start of pregnancy was determined on the basis of positive vaginal smears. There were no treatment-related effects observed in either the parents or the foetuses. The maternal, foetal and teratogenic NOAELs were 100 mg active salt/kg bw/day (highest dose tested).


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Section C2.3.7 Specific considerations

No data were identified relating to the other endpoints of toxicity of HEDP.

Section C2.3.8 Summary of human relevant health effects.

• No data relating to the potential toxicity of HEDP in humans was identified.

• In experimental species, HEDP is excreted primarily via faeces (80 %) and to a lesser extent via urine (10 %) following ingestion.

• HEDP is moderately toxic via ingestion but is of low toxicity via dermal exposure.

• HEDP is moderately irritating to the skin and moderately to severely irritating to the eye.

• HEDP disodium salt did not show any sensitisation potential and no alerts for sensitisation were identified in QSAR models.

• Repeat dose oral studies showed effects of HEDP and HEDP disodium salt on haematological parameters.

• Mutagenicity and genotoxicity studies indicate that HEDP does not have mutagenic potential in vivo.

• No carcinogenicity data were available for HEDP but HEDP sodium salt is not carcinogenic. QSAR models did not identify any alerts for carcinogenicity.

• Reproductive and developmental toxicity studies did not show adverse effects of HEDP but HEDP disodium salt induced fetotoxicity following oral exposure.

Section C3.0 Identification of relevant point(s) of


Section C3.1 Critical endpoints for human health risk assessment

purposes

The POD for use in the risk assessment (see Section C4.0) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to HEDP through any exposure route. However, a number of oral repeated dose experimental studies have been carried out for HEDP, for systemic and specific organ toxicity. These are summarised in Table C3.1.


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Table C3.1 Oral route toxicity studies for HEDP



Short term / 28 days

Rat / Wistar 0 - 100 mg/kg bw/day via oral gavage

No treatment related effects were seen. NOAEL of 100 mg/kg bw/day (highest dose tested).

Monsanto, date unknown (cited in IUCLID 2000)


Rat / strain unknown

Doses unknown via oral gavage

NOAEL of 30 mg/kg bw/day (basis of NOAEL unknown).

ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004 (cited in CIR 2016)

Subchronic / 90 days (7 days per week)

Rat / Charles River 0 -10000 ppm (0, 154, 524 and 1583 mg/kg bw/day in males and 0, 166, 545 and 1724 mg/kg bw/day in females) via the diet

At 1583 mg/kg bw/day - that bilateral mineralized microconcretions in kidney tubules and extramedullary haematopoiesis but not toxicologically relevant. No treatment related effects were seen. NOAEL of 1583 mg/kg bw/day (highest dose tested).

Industrial Biotest Labs Inc., 1979 (cited in HERA 2004; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004)

Subchronic / 90 days (7 days per week)

Rat / Charles River 180 - 1800 mg/kg bw/day via the diet

At 1800 mg/kg bw/day - decreased food intake and body weight gain and increased red blood cells in males, and decreases in white blood cells, haemoglobin and haematocrit and liver weight in females NOAEL of 600 mg/kg bw/day derived based on haematological effects.

Monsanto, 1981 (cited in IUCLID 2000; ECHA REACH registration dossier for Etidronic acid 2020)

Subchronic / 90 days

Rat / Sprague-Dawley

0, 200, 500 or 1000 ppm HEDP disodium salt (0 – 817 mg/kg bw/day for males and 0 – 1000 mg/kg bw/day for females) via the diet

At 169/195 mg/kg bw/day – decrease in red cell parameters NOAEL of 41 mg/kg bw/day derived based on haematological effects.

Huntingdon Research Centre, 1977 (cited in HERA 2004)


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Rat / Charles River 260 and 1300 mg/kg bw/day disodium HEDP salt via the diet

At 1300 mg/kg bw/day – slight increase in relative kidney weight without histopathological findings NOAEL of 260 mg/kg bw/day derived (highest dose tested).

Nixon et al., 1972 (cited in HERA 2004)

Subchronic/ 90 days (7 days per week)

Dog / beagle - 250 mg/kg bw/day via the diet

No treatment related effects were seen. NOAEL of 250 mg/kg bw/day (highest dose tested).

Monsanto, 1981 (cited in IUCLID 2000)

Subchronic/ 90 days (7 days per week)

Dog / beagle 0, 1000, 3000 and 10000 ppm (0, 191, 554 and 1746 mg/kg bw/day in males and 0, 202, 553 and 1620 mg/kg bw/day in females) via the diet

All doses - Erythrocyte counts were slightly elevated in females which was not accompanied with histopathological changes to the haematopoietic system. NOAEL of 1620 mg/kg bw/day (highest dose tested).

Industrial Biotest Labs Inc., 1975 (cited in HERA 2004; ECHA REACH registration dossier for Etidronic acid 2020; OECD SIDS 2004)

Chronic / 2 year Rat / Sprague Dawley

0, 200, 5000 or 10000 ppm HEDP disodium salt (0 – 384 mg/kg bw/day for males and 0 – 493 mg/kg bw/day for females) via the diet

At 78/96 mg/kg bw/day – anaemia was observed NOAEL of 19 mg/kg bw/day derived based on haematological effects.

Huntingdon Research Centre, 1979 cited in HERA 2004

Developmental toxicity study

Rabbit (strain unknown)

0, 25, 50 or 100 mg /kg bw/day HEDP disodium salt

No treatment related effects were seen. Maternal, foetal and teratogenic NOAEL of 100 mg/kg bw/day (highest dose tested).

Nolen and Buehler, 1971 (cited in HERA 2004)

Two-generation study

Rat / Charles River 0 – 0.5 % HEDP disodium salt (0, 112 or 447 mg active salt/kg bw/day via the diet

At 447 mg/kg bw/day – significant decrease in the number of live pups born Developmental NOAEL of 112 mg/kg bw/day derived based on fetotoxicity and reproductive NOAEL of 447 mg/kg bw/day (highest dose tested).

Nolen and Buehler, 1971 (cited in HERA 2004)



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The data indicate that haematological changes are the most sensitive effects following oral exposure to HEDP (discussed further in section C3.3). There is no evidence from currently available studies that HEDP is carcinogenic or causes reproductive or developmental toxicity.

Section C3.2 Current health-based guidance values


Section C3.3 Identification of POD to be used for the risk

assessment

The lowest effect level following repeated oral exposure to HEDP disodium salt was reported in a 2-year feeding study (Huntingdon Research Centre, 1979 cited in HERA 2004). A NOAEL of 19 mg/kg bw/day was determined based on haematological disturbances indicated by anaemia, reduction red cell parameters and pallor were observed, which were probably due to the chelating properties of HEDP. This NOAEL is considered to be the most sensitive POD.

Section C4.0 Drinking water risk assessment


Few adverse effects were seen following oral exposure to HEDP in experimental studies.


Based on the data obtained, the NOAEL is considered to be 19 mg/kg bw/day (highest dose tested). This is used as the POD.

Assessment factor (AF)



AF Justification



Conversion from LOAEL to NOAEL

1 NOAEL used as POD

Use of subchronic data 1 NOAEL based on a2-year study

Inadequate databases 1 Adequate database available

Total AF 100


Based on a POD of 19 mg/kg bw/day and an AF of 100, the proposed TDI is 190 µg/kg bw/day.




• 380 µg/ L for a 10 kg child drinking 1 L water per day


56


Exposure assessment

No measured drinking water concentrations were available for HEDP. The maximum concentration of HEDP modelled in drinking water, as reported by WRc (2014), was 20.5 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.

Based on default assumptions, the daily intake would be:

• 0.68 µg /kg bw/day for a 60 kg adult drinking 2 L water per day

• 2.05 µg /kg bw/day for a 10 kg child drinking 1 L water per day

• 3.08 µg /kg bw/day for a 5 kg infant drinking 0.75 L water per day


The maximum intake of HEDP via drinking water by adults, children and infants is less than the TDI (HQ<1) and the maximum modelled concentration of HEDP in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to HEDP via drinking water.

Risk communication

The MOEs for HEDP, based on the NOAEL of 19 mg/kg bw/day and maximum intakes of 0.68, 2.05 and 3.08 µg /kg bw/day, are 27805, 9268 and 6179 for adults, children and infants, respectively. As the MOE’s are >100 this indicates that exposures are not of concern in terms of risk to public health.

References

CIR (2016). "Safety assessment of Etidronic acid and its simple salts as used in cosmetics. Accessed Jan 2020.".

ECHA REACH registration dossier (2020). Etidronic acid. https://echa.europa.eu/registration-dossier/-/registered-dossier/16011 Accessed August 2020.

HERA (2004). Human & Environmental Risk Assessment on ingredients of European household cleaning products; Phosphonates (CAS 6419-19-8; 2809-21-4; 15827-60-8). Accessed Jan 2020.

IUCLID dataset (2000). ATMP. European Commission, ECB. Accessed Jan 2020.

IUCLID dataset (2000). HEDP. European Commission, ECB. Accessed Jan 2020.

OECD SIDS (2004). SIDS Dossier OECD HPV Chemical Programme, SIDS Dossier, approved at SIAM 18 (20/04/2004)). Etidronic acid. Accessed Jan 2020.

WRc (2014). Risk to drinking water from Personal Care Products and Domestic Cleaning Products. Accessed Jan 2020.

https://echa.europa.eu/registration-dossier/-/registered-dossier/16011



57

Section D: Diethylenetriamine

penta(methylene phosphonic acid)

(DTPMP)

Section D1.0 Chemical identification, use and


Section D1.1 Reasons for consideration

The WRc (2014) report assessed the use of DTPMP as a phosphonate in dish, laundry and household cleaning products. UK consumption decreased from 2310 tonnes/year in 2007 to 2040 tonnes/year in 2012. Levels of DTPMP in drinking water were estimated to be in the range 0.69 – 3.47 µg/L. DTPMP is included for further consideration here as although the estimated exposures through drinking water and bathing were lower than the estimated exposure through intended use (SED of 0.005 mg/kg bw/day), the difference between the estimated exposures was much less than for other chemicals. Therefore, there is the potential for significant exposure to DTPMP through the combination of direct, intended use, and subsequent secondary exposure from consumption of drinking water and bathing (WRc 2014).

Section D1.2 Identification and physicochemical properties

Being multifunctional acids, phosphonates will form salts or complexes of different composition, depending on the chemical composition and the pH of the environment (HERA, 2004). These phosphonates are primarily used as acids and sodium salts. Their behaviour in the body does not depend on the presence of sodium as the counter ion. Therefore, phosphonate salts may be used to assess the toxicity of the phosphonate acid, although it should be noted that the toxicity of the salts in some cases i.e. for HEDP is greater than for the acid. This is considered to be related to the perturbation of iron homeostasis as a result of the chelating properties of HEDP. In this case, only data for HEDP was used to assess repeat dose toxicity. However, Hera (2004) stated ‘There is, however, no need to distinguish between the acid and salt form of the respective phosphonic acid. The basic justification for this is that in dilute aqueous conditions of defined pH a salt will behave no differently to parent acid. The effect of the counter-ion (usually sodium) is not judged to be significant’.


58

Table D1.1 DTPMP identification and physicochemical properties


Cas No

15827-60-8

IUPAC name [bis(phosphonomethyl)amino]ethyl})amino]methyl}phosphonic acid

Chemical Group Phosphonates


DSL* name Phosphonic acid, [[(phosphonomethyl)imino]bis[2,1-ethanediylnitrilobis(methylene)]]tetrakis-

Common name Diethylenetriamine penta(methylenephosphonic) acid

Formula: C9H28N3O15P5

Molecular weight 573.196 g/mol

Structure

Solubility Soluble in water


Boiling point 480 °C


Log Pow -3.40 (25 °C)

Vapour Press. 0.0000013 kPa (20 °C) Sources: (WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]] tetrakisphosphonic acid 2020)

Section D1.3 Hazard classifications

DTPMP has been assigned a hazard category of Skin Irrit. 2 with hazard statement H315 – causes skin irritation, Eye Dam. 1 with hazard statement H318 – causes serious eye damage in the REACH dissemination page (ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]] tetrakisphosphonic acid 2020)

The notified EU classification and labelling according to CLP regulation (EC) No. 1272/2008, includes Eye Irrit. 2 with hazard H319 – causes serious eye irritation, Eye Dam. 1 with hazard statement H318 – causes serious eye damage, Skin Irrit. 2 with hazard statement H315 – causes skin irritation, Skin Corr. 1A, 1B and 1C with hazard statement H314 – causes severe skin burns and eye damage and STOT RE3 with hazard statement H335 – may cause respiratory irritation. There is no harmonised classification.

Section D1.4 Occurrence, production and use

Phosphonates, including DTPMP, are used in a wide range of products, including regular and compact laundry detergents, fabric conditioners, laundry additives, hand and machine dishwashing detergent and surface, carpet and toilet cleaners (HERA 2004).


59

Section D1.5 Human exposure

Exposure to DTPMP occurs predominantly during pre-treatment of laundry, through direct contract from hand washing laundry, hand dishwashing and using hard surface cleaners, indirect contact from wearing laundered clothes, inhalation of laundry powder dust and aerosol particles, and oral exposure of residues on eating utensils and dishes as well as from exposure to residues found in water (HERA 2004).

Section D2.0 Human relevant health effects

Section D2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of DTPMP in humans.

Physicochemical data (for phosphonic acid compounds (group 1 and 2), suggest that phosphonic acid compounds will not be readily absorbed from the gastrointestinal tract (OECD SIDS 2004; HERA 2004). Toxicokinetic data on DTPMP in animals are limited but confirm that oral absorption is low, being approximately 2-7 % in animals and 2-10 % in humans.

The majority of DTPMP is eliminated in the faeces after oral administration. Only negligible amounts are found in faeces, urine and in the body following dermal administration (HERA 2004).

Section D2.2 Acute toxicity

The oral acute toxicity of DTPMP was determined in Sprague-Dawley rats according to OECD 401 (Acute Oral Toxicity) (Younger Laboratories, 1971 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]]tetrakisphosphonic acid 2020); Monsanto unpublished report cited in OECD SIDS 2004). Five rats per group (2-3 per sex) were administered 2906, 3660, 4605 or 5800 mg/kg bw via oral gavage following which they were observed for signs of toxicity. All animals died in the top dose group, three females at 4605 mg/kg group and two females in the 3660 mg/kg bw group. Clinical signs of toxicity included reduced appetite and activity, slight lethargy, rapidly increasing weakness, collapse and death. On necropsy, slight liver discoloration and acute gastro-intestinal inflammation were observed. An LD50 of 4164 mg/kg was determined. Although the study was pre-GLP and was not in full compliance with OECD guideline it was still deemed to be scientifically reliable (ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]] tetrakisphosphonic acid 2020).

One acute dermal test in rabbits was identified (Younger Laboratories, 1971 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis [ethane-2,1-diylnitrilobis(methylene)]]tetrakisphosphonic acid 2020; Monsanto unpublished report cited in OECD SIDS 2003). Four rabbits (two per sex) were administered a single dermal application of 1833 2906 or 4605 mg/kg bw for 24 hours. None of the treated animals died or showed signs of toxicity. The dermal LD50 was determined to be >4605 mg/kg. The study was not in compliance with OECD guidelines and GLP regulations.


60

Section D2.3 Repeat dose toxicity

Section D2.3.1 Systemic effects

No data were identified relating to the systemic effects of DTPMP in humans or animals.

In the absence of data for DTPMP, the DTPMP salt has been used as a surrogate chemical in a read-across approach (see justification for read-across chemicals above (Section D1.2)).

In a subchronic oral feeding study carried out according to OECD 408, administration of DTPMP sodium salt (0, 100, 1000 and 10000 ppm, equivalent to 0, 8, 83 and 850 mg salt/kg bw/day for males and 0, 9.2, 92.3, and 903 mg salt/kg bw/day for females) to rats for 90 days in feed resulted in changes in haematological parameters (red blood cell levels increased, mean cell volume and haemoglobin decreased in both sexes) at the top dose. Perturbations of iron and calcium homeostasis indicated by serum iron and binding capacity as well as increased bone density, and reduced incidence of microliths were also reported in female kidneys. The latter effects were thought to be due to effects of changes in calcium homeostasis and not toxicologically significant. A NOAEL of 83 mg/kg bw/day was determined based on the haematological changes (Central laboratory, 1988 cited in HERA 2004). The study was of high quality and in compliance with GLP and OECD method 408.

A further 90-day feeding study was carried out in which Sprague Dawley rats were fed DTPMP salt (0, 4, 45 or 511 mg salt/kg bw/day for males and 0, 6, 57 or 656 mg salt/kw bw/day for females). No further information is available (Proctor and Gamble, 1978 cited in HERA 2004). Decreased body weights and liver weights in males and decreased spleen hemosiderin in both sexes were reported at the middle dose but were fully reversible and were not accompanied by histopathological changes. At the top dose, signs of anaemia were observed in both sexes (pale extremities, decreased haematocrit, decreased haemoglobin, decreased red blood cells, decreased plasma iron, heart weight and spleen hemosiderin), and in males body weights, plasma calcium and liver weights were decreased. The study authors determined a NOAEL of 4 mg salt/kg bw/day based on the decrease in male body weights and liver weights at the mid-dose. However, the HERA document considered such effects not to be toxicologically relevant due to the lack of histopathological changes and the reversibility of effects. Hence a NOAEL of 45 mg salt/kg bw/day was determined, based on haematological changes at the top dose (HERA 2004). The reliability of this study is uncertain due to the lack of data in the study report.

A chronic 1-year feeding study was also carried out, in which Fischer 344 rats were administered an aqueous solution 50 % DTPMP salt (0, 4, 20, 100 and 500 mg salt/kg bw/day) via the diet (Procter and Gamble, 1982 cited in HERA 2004). A decrease in spleen hemosiderin was observed in males at 100 mg/kg bw/day. At the highest dose, decreased haematocrit, haemoglobin, plasma iron and magnesium, liver and spleen weight and spleen hemosiderin were observed, together with unspecified changes in liver histopathology. A NOEL of 20 mg/kg bw/day was determined by the study authors based on the changes in spleen hemosiderin in males, although HERA considered such effects not to be toxicologically significant (HERA 2004). Consequently, a NOAEL of 100 mg/kg bw/day is considered more appropriate, based on effects seen at the top dose.


61

Section D2.3.2 Irritancy and corrosivity

No data were identified relating to the irritancy potential of DTPMP in humans.

A number of skin and eye irritation studies have been carried out. In a study carried out according to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) 0.5 ml aqueous solution of 50 % DTPMP was administered to intact skin of three New Zealand rabbits under an occlusive dressing for four hours (Safepharm laboratories Ltd, 1982 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]]tetrakisphosphonic acid 2020; Monsanto unpublished report cited in OECD SIDS 2003). Skin reactions were graded 1, 24, 48 and 72 hours after removal of the dressing. Mild erythema (score = 1) was observed in one animal up to 72 hours after treatment and slight oedema in one animal in the first hour after dosing. Overall, it was concluded that DTPMP is only minimally irritating.

Similar conclusions were made in other studies. In the first study, 0.5 ml aqueous solution of 50 % DTPMP was applied under an occlusive dressing for 24 hours to skin of three rabbits (Younger Laboratories, 1967 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]] tetrakisphosphonic acid 2020). After 24 hours, very slight erythema was observed in all animals, which was reversible within 48 hours. DTPMP was considered to be slightly irritating. In a second study, 0.5 ml aqueous solution of 50 % DTPMP was applied to intact and abraded rabbit skin. No further details are available. Skin erythema (score=1) was observed in all animals 24 hours after treatment which was reversible within 48 hours. DTPMP was considered to be minimally irritating to skin.

Two studies were found that investigated the irritation potential of DTPMP on the eye. In the first study, 0.1 ml of an aqueous solution of 50 % DTPMP was administered in the conjunctival sac of the right eye of three New Zealand white rabbits (Younger Laboratories, 1971 cited in HERA 2004; IUCLID 2000; Monsanto unpublished report cited in OECD SIDS 2003). Rabbits were observed for up to seven days after treatment. Moderate to severe initial pain, moderate erythema, slight oedema and moderate discharge was observed which were full reversible within seven days. DTPMP was concluded to be minimally irritating.

In a subsequent study in rabbits using the same test substance and protocol, more severe effects were reported (Younger Laboratories, 1971 cited in HERA 2004). Severe initial pain, corneal cloudiness, necrosis in conjunctival sac, slight oedema and copious discharge were all reported, with only slight improvement after seven days. Therefore, DTPMP was considered to be moderately irritating to rabbit eyes. In the same study, 0.1 ml DTPMP was administered to six rabbits for one minute, followed by rinsing. Severe initial pain, necrosis in conjunctival sac, slight oedema, copious discharge and corneal cloudiness was reported which did not completely clear by day seven. In this study, DTPMP was considered moderately irritating.

Section D2.3.3 Sensitisation

No data were identified relating to the sensitisation properties of DTPMP in humans.

DTPMP did not show any evidence of skin sensitisation in either the Buehler test or the Magnusson and Kligman guinea pig maximization test (Procter and Gamble 1976; Unilever unpublished data cited in HERA 2004). No further details are available.


62

Section D2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of DTPMP in humans.

An Ames test was carried out according to OECD Guideline 471 (Bacterial Reverse Mutation Test) (Monsanto unpublished report, 1981 cited in HERA 2004; OECD SIDS 2003, ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]] tetrakisphosphonic acid 2020), in which an aqueous solution of DTPMP (0.0065, 0.026, 0.13, 0.65, 1.95 and 6.5 µg/plate) was tested with Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 with and without metabolic activation. The test was negative as no increase in revertants was observed.

DTPMP was also tested in three in vitro mammalian cell gene mutation assays, according to OECD 476 (In Vitro Mammalian Cell Gene Mutation test). In the first study (SRI International, 1982 cited in HERA 2004; IUCLD 2000; Monsanto unpublished report cited in OECD SIDS 2003; ECHA REACH registration dossier for [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]]tetrakisphosphonic acid 2020), an aqueous solution of DTPMP (70-1400 µg/L) was added to mouse lymphoma L5178Y TK+/- plate assay, both with and without S9 metabolic activation. A dose-related increase in mutation frequency at the TK locus was seen in the presence and the absence of S9.

A second study was carried out to assess if the response was due to a change in pH, by neutralising DTPMP with NaOH. Mutations were induced by the neutralised DTPMP at the TK locus in the presence of S9, indicating that the positive response seen was not due to the change in pH.

A third study was carried out (Microbiological Associates 1983 cited in HERA 2004) in which a neutralised aqueous solution of DTPMP was tested in a standard mouse lymphoma L5178Y TK+/- plate assay (1.2-17.2 µg/L without metabolic activation and 3-9 µg/L with metabolic activation). Again, mutations were seen at the TK locus.

DTPMP was also tested in an in vitro mammalian cell gene mutation assay with CHO cells using mutations at the HPRT locus (Pharmakon Research International, 1984 cited in HERA 2004). An aqueous solution of DTPMP (>8000 µg/mL) was tested with and without metabolic activation and showed no evidence of mutagenic potential.

An in vivo cytogenicity study was also carried out with DTPMP in Sprague-Dawley rats (Monsanto, 1983 cited in HERA, 2004). Twelve animals (6/sex/group) were administered, by oral gavage, a single dose of an aqueous solution containing 19.7 % DTPMP acid (equivalent to 0, 200, 660 and 1970 mg/kg bw) which was neutralized to pH 7. Animals were sacrificed at 6, 12, 24 and 48 hours. At 1970 mg/kg bw, 25 % of the animals died, mild clinical signs were observed and both sexes showed a decrease in body weight. There was no decrease in mitotic index and no increase in chromosome aberrations in any group.

Section D2.3.5 Carcinogenicity

No data were identified relating to the systemic effects of DTPMP in humans or animals.


63

In the absence of data for DTPMP, DTPMP sodium salt has been used as the surrogate chemical for a read-across approach (see justification for read-across chemicals (Section D1.2)).

A 2-year chronic toxicity/carcinogenicity study was conducted DTPMP sodium salt (Procter and Gamble, 1987 cited in HERA 2004). A neutralised solution containing 50 % of the sodium salt, was administered via the diet to Fischer 344 rats (50/sex/dose) at doses of 0, 4, 20 and 100 mg/kg bw/day. No biologically significant differences in neoplastic findings were found between control and treated groups. The reliability of the study could not be determined due to the absence of details on the protocol and the results (HERA 2004).

QSAR modelling

Due to the absence of experimental data on DTPMP, QSAR modelling was carried out (by the authors of this report) using ToxTree, VEGA and the OECD Toolbox, to identify the presence of structural alerts for carcinogenicity.

In ToxTree, DTPMP was negative for genotoxic and non-genotoxic carcinogenicity.

In VEGA, using the CAESAR, ISS, IRFMN/Antares, IRFMN and IRFMN/ISSCAN-CGX models, DTPMP was predicted to be non-carcinogenic. However, all these predictions should be treated with caution. All models may not be reliable as DTPMP is outside the Applicability Domain of all models. There were also other issues with the prediction that makes it unreliable. These include only moderately similar compounds with known experimental value being found in the training set, similar molecules found in the training set having experimental values that disagree with the predicted value, and a prominent number of atom centered fragments of the compound not being found in the compounds of the training set, or are rare fragments.


Section D2.3.6 Reproductive and developmental toxicity

No data were identified relating to the reproductive and/or developmental toxicity of DTPMP in humans.

In a rat study, DTPMP (0, 300, 1000 and 3000 ppm) was administered via the diet to Long-Evans rats (20 per dose group) (Bio/Dynamics Inc., 1979 cited in HERA 2004). Females were treated over 2 generations and males over 1 generation. Administration of the test substance started after mating of the untreated parents on day 0 of gestation and at weaning until necropsy of F2b generation. The overall intake of DTPMP was calculated to be 0, 28, 97 or 294 mg/kg bw/day for males and 0, 32, 108 or 312 mg/kg bw/day for females. A statistically significant reduced pup body weight was observed in the F2a litters from dams in the high dose group. These changes were, however, neither not observed in the F1 litter nor replicated in the F2b litter suggesting they are unrelated to treatment. A reproductive NOAEL of 294 mg/kg bw/day was determined and a developmental NOAEL of 312 mg/kg bw/day (highest doses tested).


64

Sprague-Dawley rats (number not given) were administered of 0, 500, 1000 or 2000 mg active salt/kg bw/day DTPMP sodium salt on days 6 to 19 of gestation (Monsanto, 1982 cited in HERA 2004). Maternal toxicity indicated by a 30 % decrease in body weights were observed at the high dose level of 2000 mg/kg bw/day. No significant increase in the number of malformations was observed at any dose level. Increased incidences in vertebral anomalies, including missing, reduced or fused vertebral arches, were observed in the 1000 and 2000 mg/kg bw/day groups, but these findings were considered to be of equivocal toxicological relevance and the incidence did not differ significantly from the control. A maternal NOAEL of 1000 mg/kg bw/day a developmental NOAEL of 2000 mg/kg bw/day and a fetal NOAEL of 1000 mg active salt/kg bw/day were determined (the basis for these NOAELs was not defined). The study was performed in compliance with GLP and considered to be of good quality.

Section D2.3.7 Specific considerations

No data were identified relating to other endpoints of toxicity of DTPMP.

Section D2.3.8 Summary of human relevant health effects

• No data were identified relating to the toxicokinetics of DTPMP in humans.

• In animals, DTPMP is excreted primarily via faeces.

• DTPMP is of low acute oral and dermal toxicity.

• DTPMP is mildly irritating to the skin and eyes.

• No sensitisation reactions were seen with DTPMP.

• Few data were available for DTPMP regarding repeat dose toxicity. DTPMP salt caused changes in haematological parameters.

• Mutagenicity and genotoxicity studies indicate that DTPMP does not have mutagenic potential.

• No carcinogenicity data were available but DTPMP salt was not carcinogenic. QSAR models did not identify any alerts for carcinogenicity.

• Reproductive and developmental toxicity studies did not show adverse effects of DTPMP following oral exposure.

Section D3.0 Identification of relevant point(s) of


Section D3.1 Critical endpoints for human health risk assessment

purposes

The POD for use in the risk assessment (see Section D4.0) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to DEGBE through any exposure route. However, a number of oral repeated dose experimental studies have been carried out for DTPMP, for systemic and specific organ toxicity. These are summarised in Table D3.1.


65

Table D3.1 Oral route toxicity studies for DTPMP

Type/Duration Species/Strain Dose range (mg/kg bw/day); route of exposure


Subchronic /90 days Rat / Wistar 0 - 10000 ppm (0, 8, 83 and 850 mg/kg bw/day in males and 0, 9.2, 92.3, and 903 mg/kg bw/day in females) DTPMP salt via the diet

At top dose - changes in haematological parameters NOAEL of 83 mg/kg bw/day derived based on haematological effects.

Central Laboratory, 1988 (cited in HERA 2004)

Subchronic/ 90 days Rat / Sprague Dawley

0 - 511 mg/kg bw/day for males and 0, 6, 57 or 656 mg/kg bw/day for females DTPMP salt via feed

At top doses – anaemia (pale extremities, decreased haematocrit, decreased haemoglobin, decreased red blood cells, decreased plasma iron, heart weight and spleen hemosiderin) NOAEL of 45 mg/kg bw/day derived based on haematological effects.

Proctor and Gamble, 1978 (cited in HERA 2004)

Chronic / 1 year Rat / Fischer 344 0 - 500 mg/kg bw/day DTPMP salt via the diet

At 500 mg/kg bw/day - decreased haematocrit, haemoglobin, plasma iron and magnesium, liver and spleen weight, and spleen hemosiderin and unspecified changes in liver histopathology NOAEL of 100 mg/kg bw/day derived based on haematological effects.

Proctor and Gamble, 1982 (cited in HERA 2004)

Chronic / 2 years Rat / strain unknown

0, 50, 150 or 500 mg/kg bw/day via the diet

500 mg/kg bw/day - Reduced body weights and changes in liver, spleen and kidney weights or weight ratios which were not accompanied with histopathological findings. NOAEL of 150 mg/kg bw/day was derived

Giovannit Bozzetto S.p.A (cited in IUCLID 2000)


Rat / Sprague Dawley

0 – 2000 mg/kg bw/day DTPMP disodium salt

2000 mg/kg bw/day – decrease maternal body weight

Monsanto, 1982 (cited in HERA 2004)


66



Maternal NOAEL of 1000 mg/kg bw/day a developmental NOAEL of 2000 mg/kg bw/day and a fetal NOAEL of 1000 mg active salt/kg bw/day were derived

Two-generation study

Rat / Long-Evans 0, 300, 1000 or 3000 ppm DTPMP (0, 28, 97 or 294 mg/kg bw/day in males and 0, 32, 108 or 312 mg/kg bw/day in females) via the diet

294/312 mg/kg bw/day - A statistically significant reduced pup body weight in the F2a litters which were not observed in the F1 litter or replicated in the F2b litter hence unrelated to treatment Reproductive NOAEL of 294 mg/kg bw/day and developmental NOAEL of 312 mg/kg bw/day (highest dose tested).

Bio/Dynamics Inc., 1979 (cited in HERA 2004)


67

The data indicate that haematological changes are the most sensitive effects following oral exposure to DTPMP (discussed further in section 2.3). There is no evidence from currently available studies that DEGBE is carcinogenic or causes reproductive or developmental toxicity.

Section D3.2 Current health-based guidance values


Section D3.3 Identification of POD to be used for the risk assessment

The lowest effect level following repeated oral exposure to DTPMP salt was reported in a 90- day study by Proctor and Gamble (1978) and was associated with anaemia in female rats, resulting in a NOAEL of 45 mg/kg bw/day. The study authors had derived a NOAEL of 45 mg/kg bw/day based on haematological changes. However, confidence in the study is limited due to the absence of information on the methodology, hence the reliability of the data could not be determined.

Haematological findings were also noted in a reliable OECD 408 study in male and female Wistar rats with DTPMP salt, from which a NOAEL of 83 mg/kg bw/day was determined (Central laboratory, 1988 cited in HERA 2004). This NOAEL is considered to be the most sensitive POD.

Section D4.0 Drinking water risk assessment


The most sensitive endpoint is haematotoxicity, observed in a 90-day feeding study in rats by Proctor and Gamble (1978).


Based on the data obtained, the NOAEL is considered to be 83 mg/kg bw/day based on haematological effects. This is used as the POD.




AF Justification




1 NOAEL used as POD


Inadequate databases 1 Adequate databases

Total AF 300



68


Based on a TDI of 277 µg/kg bw/day and assuming 20 % allocation of the TDI to drinking water, the DWELs would be:


• 553 µg/ L for a 10 kg child drinking 1 L water per day


Exposure assessment

No measured concentrations in drinking water were available for DTPMP. The maximum concentration of DTPMP modelled in drinking water, as reported by WRc (2014), was 3.47 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.


• 0.12 µg /kg bw/day for a 60 kg adult drinking 2 L water per day

• 0.35 µg /kg bw/day for a for a 10 kg child drinking 1 L water per day

• 0.52 µg /kg bw/day for a 5 kg infant drinking 0.75 L water per day


The maximum intake of DTPMP via drinking water by adults, children and infants is less than the TDI (HQ <1) and the maximum modelled concentration of DTPMP in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to DTPMP via drinking water.

Risk communication

The MOEs for DTPMP, based on the NOAEL of 83 mg/kg bw/day and maximum intakes of 0.12, 0.35 and 0.52 µg /kg bw/day are 717579, 239193 and 159462 for adults, children and infants, respectively. The MOEs indicate that exposures are not of concern in terms of risk to public health.

References

ECHA REACH registration dossier (2020). [[(phosphonomethyl)imino]bis[ethane-2,1-diylnitrilobis(methylene)]]tetrakisphosphonic acid. https://echa.europa.eu/registration-dossier/-/registered-dossier/14238/1 Accessed August 2020.



IUCLID dataset (2000). DTPMP. European Commission, ECB. Accessed Jan 2020.

OECD SIDS (date unknown). Aminotri(methylenephosphonic acid). Accessed Jan 2020.

https://echa.europa.eu/registration-dossier/-/registered-dossier/14238/1

https://echa.europa.eu/registration-dossier/-/registered-dossier/14238/1

69

OECD SIDS Initial Assessment Profile (2004). DTPMP and salts (Phosphonic Acid Compounds group 3). Accessed Jan 2020.


70

Section E: Aminotris(methylene

phosphonic acid) (ATMP)

Section E1.0 Chemical identification, use and


Section E1.1 Reasons for consideration

The WRc (2014) report assessed the use of ATMP as a phosphonate in dish, laundry, and household cleaning products. UK consumption of these products decreased from 2310 tonnes/year in 2007 to 2040 tonnes/year in 2012. Levels of ATMP in drinking water were estimated to be in the range 7.15 – 57.4 µg/L. ATMP is included for further consideration here as the estimated exposures through drinking water and bathing were greater than the estimated exposure (SED of 0.005 mg/kg bw/day) through intended use (WRc, 2014).

Section E1.2 Identification and physicochemical properties.

Table E1.1 ATMP identification and physicochemical properties


Cas No

6419-19-8

IUPAC name [Bis(phosphonomethyl)aminomethyl]phosphonic acid

Chemical Group Phosphonates


*DSL name Phosphonic acid, [nitrilotris(methylene)]tris-

Common name Aminotris(methylene phosphonic acid)

Formula: C3H12NO9P3


Structure

Solubility Miscible in water.

Melting point 90.30 °C



Log Pow -3.53 (25 °C)

Vapour Press. 0.1kPa (20 °C) Sources: WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid 2019.

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Section E1.3 Hazard classifications

ATMP has been assigned a hazard category of Eye Irrit. 2, with hazard statement H319 – “causes serious eye irritation” in the REACH dissemination page (ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020).

The notified EU classification and labelling according to CLP regulation (EC) No. 1272/2008, includes Eye Irrit. 2 with hazard H319 – “causes serious eye irritation”, Eye Dam. 1 with hazard statement H318 – “causes serious eye damage”, Skin Irrit. 2 with hazard statement H315 – “causes skin irritation”, Skin Corr. 1B and 1C with hazard statement H314 – “causes severe skin burns and eye damage” and Acute Tox. 5 with hazard statement H303 – “may be harmful if swallowed”. There is no harmonised classification.

Section E1.4 Occurrence, production and use.

Although phosphonates are used in a wide range of products, ATMP is used solely in surface cleaners (HERA 2004).

Section E1.5 Human exposure

Exposure to ATMP is predominantly during pre-treatment of laundry, through direct contract from hand-washing laundry, hand dishwashing and using hard surface cleaners, indirect contact from wearing laundered clothes, inhalation of laundry powder dust and aerosol particles, and oral exposure of residues on eating utensils and dishes as well as from exposure to residues found in water (HERA 2004).

Section E2.0 Human relevant health effects

Section E2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of ATMP in humans.

In a ten-day toxicokinetics study, eight male Charles River rats were administered a single dose of 150 mg/kg bw/day 14C-ATMP by oral gavage (Hotz et al., 1995 cited in HERA 2004). ATMP was distributed throughout all bones of the body (most intense in epiphyseal plate of the long bones and in nasal turbinates). A low level of accumulation was present in stomach lining and the kidneys. Unchanged ATMP accounted for 25 % of radioactivity recovered from rat urine 0-24 h after oral administration, with 46 % present as an N-methyl derivative and 29 % as an unknown metabolite. Faecal excretion was the principal route of elimination, with 74 % of the dose being eliminated in 24 h, 83 % in 48 h and 84.4 % by day 10 following exposure. Trace amounts of radioactivity were present in urine (approx. 1 % of dose) and in blood, tissues and carcass (total approx. 0.3 %) but not in exhaled air. Overall, the mean recovery from all sources was 85.9 %. The rate of elimination of ATMP in urine, as well as whole body elimination, showed a rapid initial phase (half-life approximately 5 h) and a slower terminal phase (half-life approximately 70 h for urine and 300 h for whole body).

Section E2.2 Acute toxicity

The acute oral toxicity of ATMP has been tested in rats and mice.

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The oral acute toxicity of ATMP was determined in Sprague-Dawley rats according to OECD 401 (Acute Oral Toxicity) (Younger Laboratories, 1967 cited in HERA 2004; IUCLID 2000; ECHA 2019; Monsanto unpublished report cited in OECD SIDS date unknown). Diarrhoea, salivation and tremors were observed. On necropsy, inflammation of the gastrointestinal mucosa, and liver and kidney hyperaemia were observed. An LD50 of 2910 mg/kg was determined. Although the study was pre-GLP and was not in full compliance with OECD guidelines, it was still deemed to be scientifically reliable (ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020).

In mice, an oral LD50 of 2790 mg/kg bw was determined (Gloxhuber, unpublished data cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris (phosphonic acid) 2020). No further details are available.

One acute dermal test was identified in rabbits (Younger Laboratories, 1967 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020; Monsanto unpublished report cited in OECD SIDS date unknown). None of the treated animals died or showed signs of toxicity. The dermal LD50 was determined to be >6310 mg/kg. The study was not in compliance with OECD guideline 402 or GLP regulations.

Section E2.3 Repeat dose toxicity

Section E2.3.1 Systemic effects

No data were identified relating to the systemic effects of ATMP in humans.

In a 28-day study, 12 male rats were administered 600 mg/kg bw/day ATMP by oral gavage and no adverse effects were observed (Manley, 1981 cited in HERA 2004). A NOAEL of >600 mg/kg bw/day was determined (highest dose tested). The study was not carried out in accordance with OECD guidelines or GLP, hence quality cannot be verified (HERA 2004).

A number of feeding studies were carried out, including a dose range-finding study, a 90-day study and three 2-year studies. In a dose range-finding study preceding a 2-year oral feeding study, sixty Long-Evans rats (5/ sex/dose) were fed 0, 125, 250, 500, 750 and 1000 mg/kg bw/day ATMP over 34 days (Bio/Dynamics Inc., 1976 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020 Monsanto unpublished report cited in OECD SIDS date unknown). No treatment related effects were observed, hence a NOAEL of > 1000 mg/kg bw/d was determined (highest dose tested).

In a 90-day study according to OECD 408 (Subchronic Oral Toxicity – Rodents), Sprague-Dawley rats (number and sex unknown) were administered 6000 ppm ATMP via feed (Albright and Wilson Ltd cited in IUCLID 2000; OECD SIDS date unknown; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). No mortalities or treatment-related adverse effects were observed. A NOAEL of 6000 ppm was determined (only dose tested).

In the first 2-year feeding study, carried out according to OECD 453 (Combined Chronic Toxicity / Carcinogenicity Studies), Long-Evans rats (70/sex/dose) were administered ATMP at doses of 0, 50, 150 or 500 mg/kg bw/day (Bio/Dynamics Inc, 1979 cited in HERA 2004; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). Reduced body

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weights, changes in absolute and relative liver, spleen and kidney weights and/or weight ratios, and some erratic increases in testes and kidney weights, were observed only in the high dose group. However, these effects were minor, not consistent with time, were not consistently altered in relation to body weight, and were not accompanied by any histopathological findings. A NOAEL of 500 mg/kg bw/day was determined (highest dose tested). Although not carried out according to GLP, the study still largely followed OECD 453 and hence is considered to be reliable.

In the second 2-year feeding study, Long-Evans rats (70/sex/dose) were fed ATMP (0, 50, 150 and 500 mg/kg bw/day) in the diet (Bio/Dynamics Inc., 1979a cited in HERA 2004). Reduced body weights, changes in absolute and relative liver, spleen and kidney weights or weight ratios, and some erratic increases in testes and kidney weights were observed in the high dose group. However, these effects were not consistent with time, were minor in extent, were not consistently altered in relation to body weight and were not accompanied by any histopathological findings. There were no differences in the incidence of neoplasia in the different treatment groups (see also section 5.2.1.6). Overall, these findings were considered to be adaptive and not toxicologically relevant. In correlation with the study described above, the NOAEL was determined to be 500 mg/kg bw/day (highest dose tested). Although the study was not in compliance with GLP regulations, it conformed in all major aspects to OECD method 453 (Combined Chronic Toxicity / Carcinogenicity Studies).

In the third 2-year feeding study, rats (no., sex and strain unknown) were administered 0, 50, 150 or 500 mg/kg bw/day ATMP (Giovannit Bozzetto S.p.A cited in IUCLID 2000). Reduced body weights and changes in liver, spleen and kidney weights and/or weight ratios were observed at 500 mg/kg bw/day. No changes in histology, biochemistry, urinalysis or haematological parameters were observed. A NOAEL of 150 mg/kg bw/day was determined based on the organ weight changes. This study is considered to be less reliable as limited details about the animals tested are available and no data on the magnitude of body and organ weights are presented.

A 90-day study was also conducted with the sodium salt of ATMP. Ten Sprague-Dawley rats (5/ sex) were administered 161 mg active salt/kg bw/day for males and 175 mg active acid/kg bw/day for females) via the diet (Safepharm Laboratories Ltd., 1982 cited in HERA 2004). No treatment-related effects were observed. A NOAEL of 161 mg active acid/kg bw/day was determined (highest dose tested). The study was not in full compliance with GLP and OECD guidelines, and provided only limited information.

Section E2.3.2 Irritancy and corrosivity

No data were identified relating to the irritancy potential of ATMP in humans.

A number of skin and eye irritation studies have been carried out. In a study carried out according to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) 50 % ATMP was administered to intact skin of three New Zealand rabbits under an occlusive dressing for four hours (Safepharm laboratories Ltd, 1982 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020; Monsanto unpublished report cited in OECD SIDS date unknown). Skin reactions were graded 1, 24, 48 and 72 hours after removal of the dressing. Mild erythema (score = 1) was observed up to 24 hours after treatment. No oedema was observed.

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In a further study, undiluted ATMP powder and 25 % aqueous solution of ATMP was applied under an occlusive dressing for four hours to intact skin of six rabbits, following the Monsanto test protocol (Younger Laboratories, 1967 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). The powder did not cause any erythema or oedema, but the aqueous solution resulted in moderate erythema and oedema, which were reversible within seven days. The powder was considered to be non-irritating and the aqueous solution moderately irritating.

An acute eye irritation study was carried out in rabbits according to the Draize method (Younger Laboratories, 1967 cited in HERA 2004; IUCLID 2000; Monsanto unpublished report cited in OECD SIDS date unknown). 100 mg ATMP powder was administered to the conjunctival sac of the right eye of three rabbits. The left eye served as a control. Rabbits were observed for up to seven days after treatment. Twenty-four hours after administration, lid closure and iris congestion was observed which decreased but were still present after seven days. ATMP was considered to be moderately irritating to rabbit eyes. The study was not in compliance with GLP and OECD guidelines, but was considered to be scientifically robust and reliable (HERA 2004).

ATMP was also moderately irritating or irritating to the eyes of rabbits following 24 h exposure, based on a score of 43.6/110 and 53.6/110, respectively. In the latter study, oedema, discharge, moderate conjunctival redness and mild corneal cloudiness were reported after one hour. Eye lids were closed overnight (no further details available) (Giovanni Bozzetto S.p.A and Albright and Wilson Ltd cited in IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020).

In contrast, ATMP was not irritating in an OECD 405 (Acute Eye Irritation/Corrosion) study in New Zealand White rabbits (Albright and Wilson Ltd cited in IUCLID 2000).

Section E2.3.3 Sensitisation

There is no evidence of human sensitisation during use over 20 years (Albright and Wilson Ltd cited in IUCLID 2000; Monsanto unpublished report cited in OECD SIDS date unknown).

ATMP did not cause any skin sensitisation reactions in a poorly reported variation of the Magnusson and Kligman guinea pig maximisation test, in which twenty guinea pigs were exposed to an aqueous solution of ATMP by an initial intradermal injection. No further study details were available on the second part of the induction (i.e., topical application) or the challenge phase (Henkel KGaA, 1984 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). Due to the absence of detailed study information, the study is not considered to be reliable. However, it still gives some indication that ATMP acid does not cause skin sensitisation.

Section E2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of ATMP in humans.

ATMP was negative in a number of Ames tests. In the first Ames test, carried out according to OECD Guideline 471 (Bacterial Reverse Mutation Test), an aqueous solution of ATMP (0.0065, 0.026, 0.13, 0.65, 1.95 and 6.5 µg/plate) was tested with Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 with and without metabolic activation. The test was

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negative as no increase in revertants was observed (Monsanto unpublished report, 1981 cited in HERA 2004; OECD SIDS date unknown, ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020).

In the second Ames test, ATMP was negative when concentrations of 8 – 5000 µg/plate were tested using Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 with and without S9 metabolic activation (Albright and Wilson Ltd, cited in IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020).

ATMP was also tested in two in vitro mammalian cell gene mutation assays according to OECD 476 (In Vitro Mammalian Cell Gene Mutation test). In the first gene mutation assay (SRI International, 1982 cited in HERA 2004; IUCLD 2000; Monsanto unpublished report cited in OECD SIDS date unknown; ECHA, 2019), an aqueous solution of ATMP (up to 780 µg/L) was added to mouse lymphoma cell L5178Y TK+/- plate assay, both with and without metabolic activation. No increase in mutation frequency was seen in the absence of S9 but a dose-related positive response was observed in the presence of S9.

In the second gene mutation assay, an aqueous solution containing 50 % active acid neutralised with NaOH was tested (up to 0.78 µL/mL) in the presence of S9 only, and showed no increase in mutant frequency (SRI International, 1982 cited in HERA 2004). This result indicates that the positive result seen in the first study was an artefact due to pH perturbation and hence ATMP was not considered to be mutagenic under the test conditions.

ATMP was also tested in a micronucleus assay according to OECD 474 (Genetic Toxicology: Micronucleus Test) (Albright and Wilson Ltd, cited in IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). Male and female CFW-1 mice were administered ATMP at doses of 0, 120, 600 or 1200 mg/kg bw/day for 72 hours. No increase in the frequency of micronucleated erythrocytes occurred.

Section E2.3.5 Carcinogenicity

A 2-year combined chronic toxicity/carcinogenicity study in rats was conducted with ATMP in accordance with OECD 453 (Combined Chronic Toxicity/Carcinogenicity Study) (Bio/Dynamics Inc, 1979 cited in HERA 2004; IUCLID 2000; Monsanto unpublished report cited in OECD SIDS, date unknown; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). Long-Evans rats (70/sex/dose) were administered 0, 50, 150 or 500 mg/kg bw/day for 24 months. Similar numbers and types of tumours were observed in control and test groups. Therefore, ATMP is not considered to be carcinogenic and a NOAEL of 500 mg/kg bw/day was determined (highest dose tested). Although carried out according to OECD guideline the study was not performed in compliance with GLP.

Section E2.3.6 Reproductive and developmental toxicity

No data were identified relating to the reproductive and/or developmental toxicity of ATMP in humans.

In a three-generation study, Long-Evans rats (12 males and 24 females per group) were administered ATMP via diet at dose levels of 0, 300, 1000 or 3000 ppm (converted doses not given) (Bio/Dynamics Inc, 1979 cited in HERA 2004; IUCLID 2000; Monsanto unpublished report cited in OECD SIDS date unknown; ECHA REACH registration dossier for

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Nitrilotrimethylenetris(phosphonic acid) 2020). Treatment started 60 days prior to first mating of the F0 generation. The offspring of the first litter of each generation (F1a, F2a, F3a) were taken for necropsy on lactation day 21 (LD 21), after which the parents were re-mated and the offspring used to breed the F1b and F2b parent generations. No adverse effects were observed in either parental animals or pups at any dose level. The NOAEL for parental, reproductive and developmental effects was 275 mg/kg bw/day (highest dose tested). The study was pre-GLP and was not fully compliant with OECD guidelines. However, the study was assessed to be scientifically reliable. As the study predates current guidelines, it did not evaluate the oestrus cycle, sperm parameters or developmental milestones (HERA 2004).

ATMP (0, 100, 500 or 1000 mg/kg bw/day) was administered to pregnant Sprague-Dawley rats (number not given) by oral gavage from day 6 to 15 of gestation (Bio/Dynamics Inc., 1979 cited in HERA 2004; IUCLID 2000; Monsanto unpublished report cited in OECD SIDS date unknown; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). A slight decrease in maternal body weight gain was observed at the highest treatment rate of 1000 mg/kg bw/day. Common malformations were observed in the pup of the high dose groups, which was related to maternal toxicity. A NOAEL of 1000 mg/kg bw/day was determined (highest dose tested) and it was concluded that ATMP was not fetotoxic or teratogenic.

The same protocol was used to test neutralised ATMP in mice. Pregnant CD-1 mice (number not given) were administered 0, 100, 500 or 1000 mg/kg bw/day ATMP from day 6 to 15 of gestation (Bio/Dynamics Inc., 1979 cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). No embryonic or teratogenic effects were observed in parents or fetuses at any dose and a NOAEL of 1000 mg/kg bw/day was determined (highest dose tested).

Section E2.3.7 Specific considerations

No data were identified relating to other toxicity endpoints for ATMP.

Section E2.3.8 Summary of human relevant health effects.

• No data were identified relating to the toxicokinetics of ATMP in humans.

• In animals ATMP is excreted primarily via faeces (>80 %).

• ATMP is of moderate acute oral toxicity and low dermal toxicity.

• ATMP is mildly irritating to the skin and moderately irritating to eyes.

• No sensitisation reactions were seen with ATMP.

• ATMP caused slight changes in organ weights (i.e. spleen, liver, kidney and testes) following oral exposure in one study, but these were not considered adverse. Predominantly no effects were seen in repeat dose studies.

• Mutagenicity and genotoxicity studies indicate that ATMP does not have mutagenic potential.

• No carcinogenicity potential was seen with ATMP.

• Reproductive and developmental toxicity studies did not show adverse effects of ATMP following oral exposure.

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Section E3.0 Identification of relevant point(s) of


Section E3.1 Critical endpoints for human health risk assessment

purposes

The POD for use in the risk assessment (see Section E4.0) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to ATMP through any exposure route. However, a number of oral repeated dose experimental studies have been carried out for ATMP, for systemic and specific organ toxicity. These are summarised in Table E3.1.

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Table E3.1 Oral route toxicity studies for ATMP




Rat 600 mg/kg bw/day via oral gavage

No treatment related effects were seen. NOAEL of >600 mg/kg bw/day (highest dose tested).

Manley, 1981 (cited in HERA, 2004)


Rat / Long Evans

0 - 1000 mg/kg bw/day via the diet

No treatment related effects were seen. NOAEL of >1000 mg/kg bw/day (highest dose tested).

Bio/Dynamics Inc., 1976 (cited in HERA 2004; IUCLID 2000; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid 2020; Monsanto unpublished report cited in OECD SIDS date unknown)



6000 ppm via the diet No treatment related effects were seen. NOAEL of >6000ppm (highest dose tested).

Albright and Wilson Ltd (cited in IUCLID 2000; OECD SIDS date unknown; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020

Chronic / 2 years

Rat / Long Evans

0 - 500 mg/kg bw/day via the diet

500 mg/kg bw/day - reduced body weights, changes in absolute and relative liver, spleen and kidney weights or weight ratios, and erratic increases in testes and kidney weights (no histopathological findings). NOAEL of 500 mg/kg bw/day (highest dose tested).

Bio/Dynamics Inc, 1979 (cited in HERA 2004; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020

3-generation study

Rat / Long Evans

0, 300, 1000 or 3000 ppm via the diet

No treatment related effects were seen. Parental, reproductive and developmental NOAEL of 275 mg/kg bw/day (highest dose tested).

Bio/Dynamics Inc, 1979 (cited in HERA 2004)



0, 100, 500 or 1000 mg/kg bw/day) via gavage

1000 mg/kg bw/day - slight decrease in maternal body weight gain and malformations in pups related to maternal toxicity.


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Parental, reproductive and developmental NOAEL of 1000 mg/kg bw/day (highest dose tested).


Mouse / CD-1 0, 100, 500 or 1000 mg/kg bw/day) via gavage

No treatment related effects were seen. Parental, reproductive and developmental NOAEL of 1000 mg/kg bw/day (highest dose tested).


Shaded rows represent studies considered suitable for the risk assessment


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The data indicate that there are no significant toxicity effects following oral exposure to ATMP (discussed further in Section E2.3). There is also no evidence from currently available studies that ATMP is carcinogenic or causes reproductive or developmental toxicity.

Section E3.2 Current health-based guidance values


Section E3.3 Identification of POD to be used for the risk

assessment.

Few adverse effects were reported in rats and mice following administration of ATMP. Reduced body weights, changes in absolute and relative liver, spleen and kidney weights or weight ratios, and erratic increases in testes and kidney weights, were observed in a 2-year feeding study from which a NOAEL of 500 mg/kg bw/day was determined. However, such effects were minor, not consistent with time, were not consistently altered in relation to body weight and were not accompanied by any histopathological findings (Bio/Dynamics Inc, 1979 cited in HERA 2004; ECHA REACH registration dossier for Nitrilotrimethylenetris(phosphonic acid) 2020). Due to the lack of toxicological effects, the NOAEL is considered to be 1000 mg/kg bw/day (highest dose tested).

Section E4.0 Drinking water risk assessment


Few adverse effects were seen following oral exposure to ATMP.


Based on the data obtained, the NOAEL is considered to be 1000 mg/kg bw/day (highest dose tested). This is used as the POD.




AF Justification




1 NOAEL used as POD

Use of subchronic data 1 NOAEL based on a developmental study

Inadequate databases 1 Adequate database available

Total AF 100


Based on a NOAEL of 1000 mg/kg bw/day and an AF of 100, the proposed TDI is 10,000 µg/kg bw/day.


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• 60,000 µg/L for a 60 kg adult drinking 2 L water per day

• 20,000 µg/L for a 10 kg child drinking 1 L water per day

• 13,333 µg/L for a 5 kg infant drinking 0.75 L water per day

Exposure assessment

No measured drinking water concentrations were available for ATMP. The maximum modelled concentration of ATMP in drinking water, as reported by WRc (2014), was 20.4 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.


• 0.68 µg /kg bw/day for an adult drinking 2 L water per day

• 2.04 µg /kg bw/day for a child drinking 1 L water per day

• 3.06 µg /kg bw/day for an infant drinking 0.75 L water per day


The maximum intake of ATMP via drinking water by adults, children and infants is less than the TDI (HQ <1) and the maximum modelled concentration of ATMP in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to HEDP via drinking water.

Risk communication

The MOEs for ATMP, based on the NOAEL of 1000 mg/kg bw/day and maximum intakes of 0.68, 2.04 and 3.06 µg /kg bw/day are 1470588, 490196 and 326797 for adults, children and infants, respectively. As the MOEs are >100 this indicates that exposures are not of concern in terms of risk to public health.

References

ECHA REACH registration dossier (2020). Nitrilotrimethylenetris(phosphonic acid). https://echa.europa.eu/registration-dossier/-/registered-dossier/15198 Accessed August 2020.



OECD SIDS (date unknown). Aminotri(methylenephosphonic acid). Accessed Jan 2020.




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Section F: 2-(2-butoxyethoxy)ethanol

(DEGBE)

Section F1.0 Chemical identification, use and


Section F1.1 Reasons for consideration

A report by WRc published in 2014 assessed the risk to drinking water from the use of 2-(2-butoxyethoxy)ethanol (DEGBE) as a solvent in household cleaners and as a moisturising agent and solvent in cosmetics. Although UK consumption decreased from 2310 tonnes/year in 2007 to 2040 tonnes/year in 2012, WRc reported that the drive to make products more water soluble was likely to lead to an increase in its use in the EU. Levels of DEGBE in drinking water were estimated to be in the range 44.1 – 57.4 µg/L; the authors noted that this was based on estimated effluent concentrations of up to five times higher than measured concentrations reported in the literature. DEGBE is included for further consideration here as the estimated exposures through drinking water and bathing were found to be greater than the estimated exposure (SED of 0.0004 mg/kg bw/day) through intended use (WRc, 2014).

Section F1.2 Identification and physicochemical properties.

DEGBE belongs to the group of glycol ethers (EU RAR, 1999). Identifiers and physicochemical properties of DEGBE are outlined in Table F1.1.

Section F1.3 Hazard classifications

DEGBE has been assigned a hazard category of Eye Irrit. 2, with hazard statement H319 – “causes serious eye irritation” in the REACH dissemination page (ECHA REACH registration dossier for 2-(2-butoxyethoxy)ethanol, 2020).

The notified EU classification and labelling according to CLP regulation (EC) No. 1272/2008, includes Eye Irrit. 2, with hazard statement H319 – “causes serious eye irritation”. There is no harmonised classification.

Section F1.4 Occurrence, production and use

Glycol ethers, including DEGBE, are mainly used as solvents. DEGBE specifically is incorporated in paints, dyes, inks, detergents and cleaners to enable dissolution of components of mixtures in aqueous and nonaqueous systems. The use of DEGBE in domestic cleaning products (DCPs) is higher (37 %) than for industrial cleaning products (22 %) (ECETOC, 2005). As detailed above, the annual production of DEGBE in the EU in 2012 was 2040 tonnes/year. Nearly 60 % of this was used in cleaning agents and about 35 % in paints and surface coatings. HERA reported the maximum level of DEGBE in hard surface cleaning products to be 6 %, with ranges of DEGBE in DCPs varying between 0.01 and 4.8 % (by weight) (HERA, 2005). The use of DEGBE is also considered safe for use in cosmetic products at a maximum level of 9 % as a solvent in hair dyes (SCCS, 2006).


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Table F1.1 DEGBE identification and physicochemical properties


Cas No

112-34-5

IUPAC name 2-(2-butoxyethoxy)ethan-1-ol

Chemical Group Glycol ethers

Physical form Liquid (faint butyl odour)

DSL name Ethanol, 2-(2-butoxyethoxy)-

Common name Diethylene glycol monobutyl ether (DEGBE)

Formula: C8H18O3


Structure

Solubility Miscible in water. Very soluble in ether, alcohol and acetone, soluble in benzene

Melting point - 68 °C

Boiling point 228 – 234 °C (1013 hPa)

Density 0.948 – 0.96 (20 °C)

Log Pow 0.56

Vapour Press. 0.027 hPa (20 °C)

Sources: WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for DEGBE

Section F1.5 Human exposure

Occupational exposure is the most likely source of exposure to DEGBE in humans, with dermal and inhalation routes being key. Non-occupational exposure may occur from the use of DCPs, cosmetics containing DEGBE or through ingestion of drinking water that has been contaminated with DEGBE (EU RAR, 1999).

Section F2.0 Human relevant health effects

Section F2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of DEGBE in humans or experimental animals following exposure via the oral route or inhalation.

An in vitro dermal absorption study using human skin carried out to a protocol equivalent to OECD Guideline 428 (Skin Absorption: In Vitro Method) reported absorption of DEGBE at a slow rate (35µg/cm2/h) with a lag time of 2 h (Dugard et al., 1984).

The absorption and elimination of radiolabelled-DEGBE following 24hr dermal occluded exposure was measured in Sprague Dawley rats (male and female) using a protocol equivalent to OECD 417 (Toxicokinetics). The level of excretion of DEGBE was 97 % at 24 h, with complete


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elimination within 48 h. Excretion occurred mainly via the urine as the metabolite 2-(2-butoxyethoxy)acetic acid, with glucuronidate conjugates also being detected (up to 8 % of the total). The dermal absorption of DEGBE was found to be higher in females, with rates of 0.73 and 1.46 mg/cm2/h in males and females respectively (Boatman et al., 1993 – cited in EU RAR, 2009).

Section F2.2 Acute toxicity

SCCP reported that human exposure (route not specified) to DEGBE at a dose of 2 ml/kg (concentration not specified) was associated with cyanosis, tachypnoea, and slight uraemia (SCCP, 2006).

The acute oral toxicity of DEGBE has been tested in several experimental species using protocols equivalent to OECD Guideline 401 (Acute Oral Toxicity). In rats, LD50 values between 5660 and 7291 mg/kg in fasted animals (a higher value of 9633 mg/kg was found in non-fasted rats). An LD50 of 2410 mg/kg was determined in fasted male CD-1 mice for DEGBE (this was higher in non-fasted mice with an LD50 of 5530 mg/kg). An oral LD50 value of 2000 has been reported in the guinea pig, and values of 2200 and 2700 mg/kg in the rabbit. Sublethal effects included inactivity, laboured breathing, rapid respiration, anorexia, weakness, tremors and prostration (SCCP, 2006).

Section F2.3 Repeat dose toxicity

Section F2.3.1 Systemic effects

No data were identified relating to the systemic effects of DEGBE in humans.

30-day study, carried out prior to the introduction of GLP and OECD guidelines, has been reported in which male and female Sherman rats were administered DEGBE by oral gavage at doses of 0, 51, 94, 210, 650, 970 or 1830 mg/kg bw/day. At doses ≥94 mg/kg-day water intake was reduced and micro-pathological (unspecified) changes in liver, kidney, spleen and testis were apparent at doses ≥650 mg/kg-day (Smyth and Carpenter, 1948). Due to the limited scope of data reported by the authors, identification of a NOAEL or LOAEL from this study is not advised (US EPA, 2009).

An industry-led oral gavage study (assumed to be carried out to OECD guidelines or equivalent) of six-weeks duration has been reported in which DEGBE was administered to CD rats at doses of 0, 891, 1782 and 3564 mg/kg bw/day on 5 days per week. Clinical signs of toxicity were noted in the high dose group (dyspnoea, prostration, and unkempt hair coat, as well as bloody urine and blood around the nose and mouth in one animal), reduced body-weight gain and food consumption and histopathological changes in the spleen (congestion, red pulp hypocellularity, and hemosiderin-like pigmentation). Statistically significant (p value not cited) effects in the mid and high dose groups included haematologic indications of RBC damage in the form of decreases in red blood cell (RBC) count, haemoglobin (Hgb) and mean corpuscular haemoglobin concentration (MCHC), and increases in mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH); increased absolute and relative spleen and liver weights and lesions in the kidneys were also reported (statistical significance was not given). At all doses, hyperkeratosis in the stomach was observed; however, the authors questioned the relevance of this to humans as it was likely a result of the bolus


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exposure and direct irritant properties of DEGBE. A NOAEL of < 891 mg/kg bw/day was derived based on haematological effects (Kodak 1984 – cited in US EPA, 2009).

A 90-day study was reported in which DEGBE was administered to F344 rats at doses of 0, 70, 330 and 1630 and 0, 50, 250 and 1270 mg/kg bw/day in males and females respectively, on 5 days per week via oral gavage. Mortality was increased in a dose-dependent fashion, with numbers of surviving animals at the highest dose being too low for statistical analysis. This was considered by the authors to be due to both gavage error and DEGBE toxicity itself. Dose-related haematological effects were reported at 13 weeks as a statistically significant decrease in total white blood cell (WBC) and lymphocyte counts (p <0.05) in females but not males. In males, statistically significant (p<0.05) increases in absolute liver weight were apparent at the lowest dose, and in relative liver weight at the mid dose. In females, evidence of haemoglobinuria was noted at the highest dose (1 surviving female only) in the form of increased hyaline droplet formation in the renal tubular epithelium; in males, this was seen at all doses and not considered to be treatment related. The authors considered the lymphopoenia noted in females to be the most sensitive effect with a LOAEL of 50 mg/kg bw/day and a NOAEL of < 50 mg/kg bw/day (Hobson et al., 1987 – cited in US EPA, 2009).

Johnson et al. (2005) reported a 90-day sub-chronic study carried out to a protocol equivalent to OECD Guideline 408 (Repeated dose 90-day Oral Toxicity in Rodents) in which male and female Fischer 344 rats were administered DEGBE at doses of 0, 50, 250 or 1000 mg/kg bw/day in drinking water. The authors reported that there were no clinical signs of treatment-related toxicity. At the highest dose, treatment-related effects were observed in males and females, including decreased water and food consumption with concomitant reduction in body weight. These effects were not noted in the mid and low dose groups. At the highest dose, statistically significant increases were noted in relative liver weight and levels of hepatic cytochrome P450s and UGT (p=0.05) and decreases (p=0.05) in serum total protein, cholesterol and aspartate aminotransferase. At the mid and high doses, RBC count and Hgb levels were statistically significantly (p<0.05) reduced in a dose-dependent manner, however there were no changes to RBC indices or morphology; WBC parameters were unaffected. Johnson et al. (2005) also noted treatment-related hepatocellular hypertrophy (described as ‘very slight’) and foci of necrotic cells in the centrilobular region of the liver in a number of females (6/10) at the highest dose. Histopathology was otherwise unremarkable. In males, no treatment related effects on sperm characteristics or reproductive tissue were observed. The authors noted that the most sensitive adverse effects of DEGBE were haematological and that although statistically significant, those seen at the mid dose of 250 mg/kg bw/day were within historical control ranges. A NOAEL of 250 mg/kg bw/day was proposed by the authors based on haematological effects in both sexes (Johnson et al., 2005).

Section F2.3.2 Irritancy and corrosivity

No data were identified relating to the irritancy potential of DEGBE in humans.

In a study carried out to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) DEGBE showed slight skin irritation in the rabbit but did not reach the criteria for classification under CLP regulation (Boatman and Knaak., 2001 – cited in SCCP, 2006).


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An acute eye irritation study was reported in the rabbit (not carried out to OECD equivalent guidelines but considered acceptable for evaluation purposes) in which DEGBE resulted in moderate irritation (Ballantyne, 1984 – cited in SCCP, 2006). DEGBE does not meet the criteria for classification under the CLP regulation.

Section F2.3.3 Sensitisation

No data were identified relating to the sensitising potential of DEGBE in humans.

DEGBE was reported to be a non sensitising agent in a guinea pig maximisation test carried out using a protocol similar to OECD Guideline 406 (Skin Sensitisation) (Unilever, 1984 – cited in SCCP, 2006).

Section F2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of DEGBE in humans.

The cytogenic potential of DEGBE has been assessed in an in vivo study carried out using a protocol similar to OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test). A single maximum tolerated dose of 3300 mg/kg/bw administered to male and female mice was not associated with an increase in the incidence of micronucleated polychromatic erythrocytes at either 24, 48 or 72 hr post exposure (Gollapudi et al., 1993). Thompson et al. also reported that DEGBE did not induce sex-linked recessive lethality in Drosophilia melanogaster (Thompson et al., 1984).

In vitro studies have also been carried out to assess the mutagenic potential of DEGBE (Thompson et al., 1984). When tested with Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538, DEGBE was negative in the absence of S9 with both negative and positive results being reported in the presence of S9. Gollapudi et al. (1993) reported weakly positive results for mutagenicity, in the absence of S9, for DEGBE in mouse lymphoma cells L5178Y TK+/- at doses between 0.42 – 7.5 µl/ml, and negative results in CHO cell HPRT locus at doses between 100-5000 µg/ml.

It can be concluded from the available evidence that DEGBE does not have mutagenic potential in vivo.

Section F2.3.5 Carcinogenicity

No data relating to the carcinogenic potential of DEGBE in humans or experimental species were identified.

QSAR modelling

QSAR modelling was carried out (by the authors of this report) using ToxTree, VEGA and the OECD Toolbox.

In ToxTree, DEGBE was negative for genotoxic and nongenotoxic carcinogenicity. In VEGA, DEGBE was assessed using the CONSENUS, CAESAR (2.1.13), SarPy/IRFMN, ISS, KNN/Read-Across, CAESAR (2.1.9), IRFMN/Antares, IRFMN and IRFMN/ISSCAN-CGX models. DEGBE was predicted to be non-mutagenic, with good reliability, which is in line with experimental findings. For carcinogenicity models where DEGBE was inside the Applicability Domain, i.e.


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those of good reliability, results predicted the chemical to be non-carcinogenic. In the OECD Toolbox, there were no alerts for DNA binding or DNA alerts for AMES, chromosomal aberrations or micronuclei.

Section F2.3.6 Reproductive and developmental toxicity

No data relating to the reproductive and/or developmental toxicity of DEGBE in humans were identified.

In a study carried out using a protocol similar to OECD Guideline 414 (Prenatal Development Toxicity Study), the reproductive and developmental toxicity of DEGBE was assessed in female CD rats administered doses of 0, 250, 500 and 1000 mg/kg bw/day by oral gavage for 14 days prior to mating until GD 13 or day 21 of lactation (Nolen et al., 1985 – cited in EPA, 2009). Untreated males were mated with treated females and treated males with untreated females. The only statistically significant treatment related effect noted by the authors was reduced body weight of pups from females administered the highest dose of DGEBE, at only one time point. A decrease in the number of liveborn pups at the highest dose was seen but did not reach statistical significance. There was no evidence of parental toxicity or fertility effects in males given DEGBE from 60 days prior to and through to the end of mating. The authors derived a NOAEL for neonatal toxicity of 500 mg/kg bw/day and for parental/reprotoxicity of 1000 mg/kg bw/day (Nolen et al., 1985 – cited in EPA, 2009).

A study has also been reported in which pregnant Wistar rats were administered DEGBE in the diet at levels of 0, 25, 115 or 633 mg/kg bw/day during GD 0 – 20, using a protocol similar to OECD Guideline 415 (One-Generation Reproduction Toxicity Study); numbers of animals per group were lower than recommended in the guideline (Ema et al., 1988 – cited in EPA, 2009). Uterine and fetal examinations were undertaken at GD 20 in some females and the remainder allowed to deliver spontaneously, with pups reared until 10 weeks of age. Maternal toxicity was reflected at all doses as reduction in body weight gain (statistical significance not cited), but this was not a dose-related trend. No developmental toxicity or teratogenicity was seen, and a NOAEL of 633 mg/kg bw/day was proposed by the authors for both maternal and developmental toxicity (Ema et al., 1988 – cited in EPA, 2009).

In a GLP compliant study, pregnant Swiss CD-1 mice were administered DEGBE (in corn oil) at doses of 0 or 500 mg/kg bw/d by oral gavage on gestation days (GD) 7 to 14 and allowed to litter. Maternal toxicity was not apparent and litter viability was unaffected. Pup postnatal survival, weight gain over 3 days and birth weight were also unaffected. No examinations of pups for malformations or skeletal abnormalities was carried out (Schuler et al., 1984). A NOAEL of 500 mg/kg bw/day can be derived from this study for maternal and developmental toxicity.

Section F2.3.7 Specific considerations

The 90-day repeat dose drinking water study in Fischer 344 rats reported in section 2.3.1. also evaluated several neurotoxicity endpoints. The authors carried out detailed clinical observations, grip strength measurement, motor activity testing and sensory evaluations. No neurotoxic effects were apparent (Johnson et al, 2005).


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Section F2.3.8 Summary of human relevant health effects

• Acute human exposure at a dose of 2 ml/kg (dose and route not specified) resulted in cyanosis, tachypnea, and slight uremia. No further data relating to the potential toxicity of DEGBE in humans was identified.

• In experimental species, DEGBE is excreted primarily via urine with the major metabolite being 2-(2-butoxyethoxy)acetic acid (BEAA) (dermal exposure study only).

• DEGBE is moderately irritant to the eye and slightly irritating to the skin.

• Mutagenicity and genotoxicity studies indicate that DEGBE does not have mutagenic potential in vivo.

• There are no carcinogenicity data for DEGBE in humans or animals. Predicted to be non-carcinogenic using QSAR modelling.

• Repeat dose oral studies showed effects of DEGBE in liver, spleen, kidneys and on haematological parameters.

• Reproductive and developmental toxicity studies did not show adverse effects of DEGBE following oral exposure.

Section F3.0 Identification of relevant point(s) of


Section F3.1 Critical endpoints for human health risk assessment

purposes

The POD for use in the risk assessment (see Section F4.0) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to DEGBE through any exposure route. However, a number of oral repeated dose experimental studies have been carried out for DEGBE, for systemic and specific organ toxicity. These are summarised in Table F3.1.


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Table F3.1 Oral route toxicity studies for DEGBE



Repeated dose / 30 days Rat / Sherman 51 – 1830 via drinking water

At doses ≥94 mg/kg bw/day - water intake was reduced. At doses ≥650 mg/kg bw/day micro-pathological (unspecified) changes in liver, kidney, spleen and testis were apparent. Limited reporting does not allow NOAEL or LOAEL determination.

Smyth and Carpenter, 1948

Repeated dose / 6 weeks (5 days per week)

Rat / CD 891 – 3564 via oral gavage

At doses ≥891 mg/kg bw/day - hyperkeratosis of the stomach was apparent. Considered to be due to direct irritant properties of DEGBE. At doses ≥1,782 mg/kg bw/day - haematologic effects (reduced erythrocyte count, haemoglobin concentration, and MCHC and increased MCV) were apparent. NOAEL of <891 mg/kg bw/day derived based on haematological effects.

Kodak, 1984 (cited in US EPA, 2009)

Repeated dose/ 90 days (5 days per week)

Rat / F344 70 – 1630 (males) 50 – 1270 (females) via oral gavage

At doses of ≥50 mg/kg bw/day in females - decreased total WBC and lymphocyte counts and MCHC At doses of ≥250/330 mg/kg bw/day – mortality was apparent in males and females (authors state that some deaths due to gavage error). LOAEL of ≥ 50 mg/kg bw/day derived based on lymphopenia in females.

Hobson et al., 1987 (cited in US EPA, 2009)

Repeated dose/ 90 days (OECD 408 equivalent)

50 – 1000 via drinking water

At doses ≥250 mg/kg-day - decreased RBC count, haemoglobin, and haematocrit. Note: changes were within

Johnson et al., 2005


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historical control levels at a dose of 250 but not 1000 mg/kg bw/day. At doses of 1000 mg/kg bw/day – liver changes (increases in organ weight and hepatic cytochrome P450s and UGT levels, decreases in serum total protein, cholesterol, and serum AST, and hepatocyte hypertrophy and individual hepatocyte degeneration). NOAEL of 250 mg/kg bw/day derived based on reduced RBC count and haemoglobin in both sexes.

Prenatal Development study /60 days for males and 14 days prior to mating to GD 13 or until weaning of the offspring for females.

250 – 1000 via oral gavage

At a maternal dose of 1000 mg/kg bw/day - reduced body weight of pups during the last week of lactation. Parental or reproductive toxicity (male or female) was not apparent in any dose group. NOAEL of 500 mg/kg bw/day for neonatal toxicity was derived.

Nolen et al., 1985 (cited in EPA, 2009).

One Generation Reproduction Toxicity Study / GD 0 – 20.

25 – 633 via the diet At all doses - reduced maternal body weight gain during pregnancy was noted but this was not related to dose. No prenatal or postnatal developmental toxicity was apparent in any dose group. NOAEL of 633 mg/kg bw/day was derived for maternal and developmental toxicity.

Ema et al., 1988 (cited in US EPA, 2009)

Developmental Toxicity / GD 7 – 14.

500 via oral gavage At a dose of 500 mg/kg bw/day - maternal toxicity was not apparent.

Schuler et al., 1984.


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At a dose of 500 mg/kg bw/day - litter viability was unaffected, pup postnatal survival, weight gain over 3 days and birth weight were also unaffected. No examinations of pups for malformations or skeletal abnormalities was carried out (Schuler et al., 1984). NOAEL of 500 mg/kg bw/day can be derived for maternal and developmental toxicity.



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The data indicate that haematological changes are the most sensitive effects following oral exposure to DEGBE (discussed further in section F3.3). There are no carcinogenicity studies to assess long-term effects. There is no evidence from currently available studies that DEGBE causes reproductive or developmental toxicity.

Section F3.2 Current health-based guidance values

Type of HBGV HBGV European Commission Indicative occupational exposure limit values

(IOELV) – long term exposure limit (8hr TWA) 10 ppm (67.5 mg/m3)

European Commission Indicative occupational exposure limit values (IOELV) – short term exposure limit (15 min)

15 ppm (101.2 mg/m3)

European Commission Cosmetics regulation, Annex III Maximum of 9% in hair dye; not to be used in aerosols

UK HSE Maximum exposure limit (MEL) 120 mg/m3

US EPA Subchronic Reference Dose (RfD) 0.3 mg/kg bw/day

US EPA Chronic Reference Dose (RfD) 0.03 mg/kg bw/day

Section F3.3 Identification of POD to be used for the risk

assessment

Hobson et al. (1987) reported the lowest effect level following repeated oral exposure to DEGBE to be associated with leukopenia and lymphopenia in female rats, with a LOAEL of ≥ 50 mg/kg bw/day. However, confidence in the findings is limited by the high mortality caused by gavage accidents at the highest dose, and inconsistencies in presentation of some reported haematological effects over the duration of the study. In addition, in the studies by Johnson et al. (2005) and Kodak (1984), which used the same strain of rats at equivalent or higher doses, no leukopenia or lymphopenia were apparent.

Consistent haematological findings relating to decreased RBC counts and haemoglobin levels have, however, been recorded following oral exposure to DEGBE. The lowest effect level for these endpoints was reported by Johnson et al. (2005), with a NOAEL of 250 mg/kg bw/day in both male and female animals.

The US EPA (2009) carried out benchmark dose modelling on the data reported by Johnson et al. (2005). The polynomial, power, and Hill models were applied for RBC counts in the male rat, with a benchmark response of 1 standard deviation from the control mean (in the absence of a biologically relevant response). All models provided an adequate fit to the data, with BMDLs of 81, 280 and 328 mg/kg bw/day (males only) determined. The US EPA considered that the lowest BMDL of 81 mg/kg bw/day (based on changes in RBC in male rats) provides the most sensitive POD. This will be used for the drinking water risk assessment (Section 4). It should be noted that the BMDL is below the level of the NOAEL, which could reflect a small sample size.

Section F4.0 Drinking water risk assessment


Few adverse effects were seen following oral exposure to DEGBE in experimental animals.


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Based on the data obtained, the BMDL is considered to be 81 mg/kg bw/day. This is used as the POD.




AF Justification



Use of subchronic data 3 BMDL based on a 90 day study


Total AF 600








Exposure assessment

No measured drinking water concentrations were available for DEGBE. The maximum concentration of DEGBE modelled in drinking water, as reported by WRc (2014), was 57.4 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.


• 1.91 µg /kg bw/day for a 60kg adult drinking 2 L water per day

• 5.74 µg /kg bw/day for a 10kg child drinking 1 L water per day

• 8.61 µg /kg bw/day for a 5kg infant drinking 0.75 L water per day


The maximum intake of DEGBE via drinking water by adults, children and infants is less than the TDI (HQ<1) and the maximum modelled concentration of DEGBE in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to HEDP via drinking water.

Risk communication

The MOEs for DEGBE, based on the NOAEL of 81 mg/kg bw/day and maximum intakes of 1.91, 5.74 and 8.61 µg /kg bw/day, are 42334, 14111 and 9872 for adults, children and infants,


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respectively. As MOEs are > 100 this indicates that exposures are not of concern in terms of risk to public health.

References

Ballantyne B. Eye irritancy potential of diethylene glycol monobutyl ether. J Toxicol, Cut. Ocular Toxicol. 3: 7-16, 1984.

Boatman R, Schum D, Guest D, Stack CR. (1993). Toxicology of diethylene glycol butyl ether 2. Disposition studies with 14C-diethylene glycol butyl ether acetate after dermal application to rats. J. Am. Coll. Toxicol. 12(2), 145-154.

Department of Health and Human Services; Results of Testing Fifteen Glycol Ethers in a Short-Term In Vivo Reproductive Toxicity Assay With Attachments, EPA Doc. 408385037, Fiche No. OTS0521552

Dugard PH, Walker M, Mawdsley SJ, Scott RC. (1984). Absorption of some glycol ethers through human skin in vitro. Env Health Perspectives, 57, 193-197.

ECHA REACH registration dossier (2020). 2-(2-butoxyethoxy)ethanol). https://echa.europa.eu/registration-dossier/-/registered-dossier/15952/2/1 Accessed August 2020.

Ema M, Itami T, Kawasaki H. Teratology study of diethylene mono-n-butyl ether in rats. Drug Chem Toxicol 11: 97-111, 1988.

Gollapudi BB, Linscombe VA, McClintock ML, Sinha AK, Stack CR. Toxicology of diethylene glycol monobutyl ether. 3. Genotoxicity evaluation in an in vitro gene mutation assay and an in vivo cytogenetic test. J Am Coll Toxicol 12: 155-159, 1993.

Hardin BD, Schuler RL, Burg JR, Booth GM, Hazelden KP, MacKenzie KM, Piccirillo VJ, Smith KN. Evaluation of 60 chemicals in a preliminary developmental toxicity test. Ter Carc Mut 7: 29-48, 1987.

Johnson, K.A., P.C. Baker, H.L. Kan et al. 2005. Diethylene glycol monobutyl ether (DGBE): Two- and thirteen-week oral toxicity studies in Fischer 344 rats. Food Chem. Toxicol. 43:467–481.

Krasavage WJ, Vlaovic MS. Comparative toxicity of nine glycol ethers: III. Six weeks repeated dose study. Unpublished data, Corporate Health and Environment Laboratories, report No TX-82-06, March 15, 1982.

Microbiological Associates, Inc.; Salmonella/Mammalian-Microsome Mutagenicity Assay (Ames Test), EPA Document No. 40-8478090, Fiche No. OTS0507511, 1982.

Nolen GA, Gibson WB, Benedict JH, Briggs DW, Scardein JL. Fertility and teratogenic studies of diethylene glycol monobutyl ether in rats and rabbits. Fund Appl Toxicol 5: 1137-1143, 1985.

https://echa.europa.eu/registration-dossier/-/registered-dossier/15952/2/1


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Schuler RL, Hardin BD, Niemeier RW, Booth G, Hazelden K, Piccirillo V, Smith K. 1984. Results of testing 15 glycol ethers in a short term in vivo reprotoxicity assay. Env Health Perspectives, 57, 141-146.

Smyth HF, Carpenter CP, Further experience with the range-finding test in the industrial toxicology laboratory. J Ind Hyg 30: 63-68, 1948.

Thompson ED, Coppinger WJ, Valencia R, Iavicoli J. Mutagenicity testing of diethylene glycol monobutyl ether. Environ Health Perspect 57: 105-112, 1984.

Unilever. Bacterial reverse gene mutation assay with butyl carbitol. Research Report ULR/105D. Unilever Research, UK, 1984.

Unilever. Bacterial reverse gene mutation assay with butyl carbitol. Research Report ULR/105C. Unilever Research, UK, 1984.

Unilever, Magnusson and Kligman guinea pig maximization test with butyl dioxitol. Research Report SSM 84 369, Unilever Research, UK, 1984.


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Section G: Linear alkylbenzene

sulphonate (LAS)

Section G1.0 Chemical identification, use and


Section G1.1 Reasons for consideration

The WRc (2014) report assessed the use of LAS as an anionic surfactant in phosphonate detergents in dish, laundry and household cleaning products. UK consumption increased from 48782 tonnes/year in 2007 to 50530 tonnes/year in 2012. Levels of LAS in drinking water were estimated to be in the range 46.8 – 1030 µg/L. LAS is included for further consideration here as although the estimated exposures through drinking water and bathing were lower than the estimated exposure (SED of 0.042 mg/kg bw/day) through intended use, the difference between the estimated exposures was much less than for other chemicals. Therefore, there is the potential for significant exposure to LAS through the combination of direct, intended use, and subsequent secondary exposure from consumption of drinking water and bathing (WRc, 2014).

Section G1.2 Identification and physicochemical properties

Table G1.1 LAS identification and physicochemical properties


Cas No 68411-30-3

IUPAC name Benzene sulfonic acid, C10-C13 alkyl derivatives, sodium salt

Chemical Group Anionic surfactants


DSL name Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts

Common name Sodium LAS

Formula: C9H28N3O15P5


Structure

Solubility Soluble in water



Density g/cm3 (20 °C)

Log Pow 3.32 (25 °C)

Vapour Press. 1.3 mm Hg (20 °C)


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Sources-WRc 2014; PubChem; ChemID Plus; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020

Section G1.3 Hazard classifications

LAS has been assigned a hazard category of Skin Irrit. 2 with hazard statement H315 – “causes skin irritation”, Eye Dam. 1 with hazard statement H318 – “causes serious eye damage” in the REACH dissemination page (REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020).

The notified EU classification and labelling according to CLP regulation (EC) No. 1272/2008, includes Acute Tox. 4 with hazard 302 – “harmful if swallowed”, Skin Irrit. 2 with hazard statement H315 – “causes skin irritation” and Eye Dam. 1 with hazard statement H318 – “causes serious eye damage”. There is no harmonised classification.

Section G1.4 Occurrence, production and use

Linear alkylbenzene sulphonate is an anionic surfactant. It is used in household detergents, such as laundry powders, laundry liquids, dishwashing products and all-purpose cleaners. Minor uses include textile and fibres, chemicals, and agriculture (HERA 2013).

Section G1.5 Human exposure

Exposure to LAS is predominantly through: direct skin contact from hand washing laundry, laundry tablets, pre-treatment of clothes, hand dishwashing, cleaning sprays, liquid cleaning products (oven cleaners, bathroom cleaners, floor cleaners) and laundry pre-treatment products (spray and liquid spot removers); indirect skin contact from wearing clothes; inhalation exposure from hand dishwashing, from detergent dust during the washing process, cleaning sprays and laundry pre-treatment products (spray spot removers), liquid cleaning products (oven cleaners, bathroom cleaners, floor cleaners); accidental product ingestion, or indirectly from drinking water (HERA 2013).

Section G2.0 Human relevant health effects

Section G2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of LAS in humans. Studies with isolated human skin preparations showed that penetration through skin and subsequent systemic absorption of LAS does not occur to any significant extent at 24 to 48 h (HERA 2013).

Following administration of an aqueous solution of 35S-LAS, 80-90 % was readily absorbed from the gastrointestinal tract. Most of the absorbed 35S was eliminated within 72 hours, with 60-65 % being eliminated in the urine and 35 % being excreted in the bile and reabsorbed completely from the gastrointestinal tract. Sulfophenyl butanoic and sulfophenyl pentatonic acid were identified as major metabolites in the urine and were not reabsorbed from the kidney tubules. Although the metabolites in the bile were not identified, authors showed that no unchanged LAS was eliminated via the bile (Michael, 1968 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020).


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In rats orally dosed with 14C-NaLAS, radioactivity was detected 0.25 h after administration, and reached a maximum after 2 h. The biological half-life was 10.9 h. 14C-NaLAS was distributed to the digestive tract and bladder 4 hours after administration, as well as to the liver, kidney, testis, spleen and lung. 47 % of the radioactivity was excreted in the urine and 50 % in the faeces 68 h after administration (HERA 2013). In Rhesus monkeys administered a single oral dose of 150 mg/kg 14C-LAS (Cresswell et al.,1978 cited in HERA 2013), the concentration of radioactivity in plasma reached a maximum after 4 h and then declined during the following 6-24 h. The half-life was approximately 6.5 h. After seven consecutive daily doses of 30 mg/kg, the plasma concentrations and the biological half-life were similar to those observed after a single dose. Two hours after the last dose, the highest distribution of radioactivity was observed in the stomach, followed by the intestinal tract, kidneys, liver, lung, pancreas, adrenals and pituitary. At 24 h, concentrations were highest in the intestinal tract, indicative of biliary excretion. No accumulation of LAS occurred as the concentrations in the tissues were generally lower than plasma.

A single topical dose of 35S-LAS was administered onto the back skin of rats and guinea pigs. In the guinea pigs, 0.1 % of the total administered dose was excreted in urine. In rats, "relatively large amounts" of radioactivity were measured in the liver and kidneys. The authors concluded that there was no accumulation in specific organs and that LAS was quickly excreted in the urine after being metabolised (Chikara Debane, 1978 cited in HERA 2013).

LAS was not detected in the uterus of pregnant ICR mice following administration of a single oral dose of 350 mg/kg bw LAS on day 3 of gestation (HERA 2013).

Howes (1975) demonstrated that penetration through skin and systemic absorption of LAS does not significantly occur at 24 to 48 h in isolated human skin preparations. Similarly, in rats no radioactivity was detected in urine or faeces following percutaneous administration of 14C-LAS applied to the clipped dorsal skin of rats and washed after 15 min (Howes, 1975 cited in HERA, 2013).

Section G2.2 Acute toxicity

A number of acute toxicity tests are available in rats and mice. The oral acute toxicity of LAS was determined in rats (strain unknown) according to OECD 401 (Acute Oral Toxicity) (Huntingdon, 1984 cited in HERA 2013; US EPA 2013). Clinical signs of toxicity included piloerection, hunched posture, abnormal gait (waddling), lethargy, decreased respiratory rate, ptosis, pallor of the extremities and diarrhoea. Autopsy of rats that died revealed isolated cases of pallor of the kidneys or spleen. An LD50 of 1980 mg/kg was determined.

Other OECD 401 study (Acute Oral Toxicity) were carried out in male and female Wistar rats from which LD50 values of 2760, 2190 and 1600 mg/kg were determined. Piloerection, diarrhoea, squatting attitude, diuresis, ataxia, nose bleeding, ataxia, tremor, staggering and slight sedation occurred 30-60 minutes following administration, which subsided in surviving animals after 3-6 days (unnamed study report, 1984a, b, c cited in REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020).


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In another oral acute toxicity study, groups of 5 male and female rats were orally exposed via oral gavage to 0, 1075, 1220, 1360, or 1710 mg/kg bw LAS (Murmann, 1984 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013; OECD SIDS 2005). All animals showed some signs of toxicity within 30 minutes after administration, including diarrhoea, squatting attitude, breathing difficulties, nose bleeding, ataxia, and lethargy. Symptoms had disappeared in surviving animals by 120 h. Mortality was seen at all dose levels, with 4 of 10 animals at the lowest dose level dying. All animals at the highest dose level died. The acute oral LD50 was 1080 mg/kg bw.

A number of acute dermal toxicity tests were identified. In a limit test in rats, performed according to OECD 402 (Acute Dermal Toxicity), five male and female rats were administered 2000 mg/kg LAS (47 %) onto clipped skin under occlusive dressing for 24 h. No deaths and no systemic toxicity were observed. The LD50 was >2000 mg/kg (Kynoch, 1986 cited in HERA 2013; US EPA 2013; Unnamed study, 1986 cited in REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; OECD SIDS 2005).

Three rabbit studies were carried out by Monsanto, 1972 (cited in HERA 2014; US EPA 2013; OECD SIDS 2005). Toxicity was seen at 251-794 mg/kg bw. Clinical observations included reduced appetite, reduced activity, increased weakness and collapse. Necropsy findings consisted of haemorrhagic lungs, liver discoloration, enlarged gall bladder, and gastro-intestinal inflammation in animals that died. These studies had methodological issues as only 1 animal was used per dose, no control groups were used, and different sex animals were used.

A limit test was carried out in rats according to OECD 402 (Acute Toxicity Study), using a single dermal application of 2000 mg/kg bw LAS as 47 % of active matter (Huntingdon, 1986 cited in HERA 2013). No deaths occurred and no systemic toxicity was reported. The dermal LD50 is > 2000 mg/kg bw.

Section G2.3 Repeat dose toxicity

Section G2.3.1 Systemic effects

A number of repeated dose studies have been carried out. LAS was administered to male/female CRJ-SD rats (12/dose) by oral gavage for 28 days at doses of 125, 250 or 500 mg/kg bw/day (Ito et al.,1978 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991). Soft stools and a decrease in body weight gain were observed in both sexes at all doses, and differences in serum biochemistry were seen in the mid and top dose groups compared to controls. The relative liver weight of female animals was decreased at 125 mg/kg bw but significantly increased at 500 mg/kg bw. A decrease in the relative weight of the heart, kidney and thymus was observed at 125, 250 and 500 mg/kg bw, respectively. In males, there was an increase in relative weight of adrenal glands, testes and brain at 500 mg/kg bw and a decrease in the weight of spleen and heart at 500 and 250 mg/kg bw/day, respectively. A NOAEL of 125 mg/kg bw/day was determined based on decreased body weight and body weight gain, clinical chemistry and organ weight ratios.

Fifty and 250 mg/kg bw/day LAS was administered to male and female rats (15/dose/sex; strain unknown) for 90 days in the diet. No adverse effects were reported apart from a slight


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increase in liver weight in females in the top dose group (Oser, 1965 cited in HERA 2013). The NOAEL was reported to be 50 mg/kg bw/day.

In another 90-day study, 0.02, 0.1 or 0.5 % LAS (8.8, 44 or 200 mg/kg bw/day) was administered to male and female rats (10/dose; strain not given) in the diet. No adverse effects were reported and a NOAEL of 220 mg/kg bw/day was determined (highest dose tested) (Kay et al., 1965 cited in HERA 2013).

LAS (100 ppm; 20 mg/kg bw/day) was also administered to mice (no details available) for six months in drinking water (Watari et al., 1977 cited in HERA 2013; Soap and Detergent Association 1991). Hepatotoxicity was reported at one and six months. Some effects were still evident after a two-month recovery period whilst cellular effects had reversed. A LOAEL of 20 mg/kg bw/day was determined (the only dose tested).

LAS (0.07, 0.2, 0.6 or 1.8 % (40, 115, 340 or 1030 mg/kg bw/day) was administered to rats (10/dose/sex) for six months in the diet (Yoneyama et al.,1972 cited in HERA 2014; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). At 115 mg/kg bw/day, male rats had increased weight of the cecum and slight degeneration of the renal tubules, and females had decreased relative kidney weights. Alterations in haematological and clinical biochemistry parameters were also observed. A NOAEL of 40 mg/kg bw/day was determined based on increased cecum weight and slight degeneration of renal tubules.

Yoneyama et al. (1976) carried out 9-month feeding and drinking water study using Wistar JCL rats (Yoneyama et al.,1976 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013; OECD SIDS 2005).

In the feeding study (Yoneyama et al., 1976), male and female rats (8/dose/sex) were administered 0.6 or 1.8 % (260/300/500 or 780/900/10007 mg/kg bw/day) in the diet for 9 months. At the lowest dose, liver weight increased in both sexes (% not reported) and a decrease in LDH in the liver and acid phosphatase in the kidneys was reported. At the high dose, body weight gain was decreased (% not reported), cecum weights were increased in males, liver and cecum weights were increased in females, and changes in liver and kidney enzymes were observed. HERA determined a NOAEL of 260 mg/kg bw/day based on decreased body weight gain and changes in clinical biochemistry. In contrast, the REACH dossier considered the lowest dose of 300 mg/kg bw/day to represent a LOAEL, based on clinical signs, body weight gain, clinical chemistry, haematological signs and change in organ weights (HERA, 2013).

In the drinking water study (Yoneyama et al., 1976) male and female rats (8/dose/sex) were administered 0.07, 0.2 or 0.6 % (85, 145 or 430 mg/kg bw/day) in drinking water for 9 months. At the mid dose there was a decrease in lactate dehydrogenase and Na, K-ATPase, and at the high dose a significant decrease in body weight gain (% not given) was seen in males as well as changes in liver and kidney enzymes (HERA, 2013; US EPA, 2013). HERA determined a

7 US EPA cited 500 and 1000 mg/kg bw/day, REACH dossier cited 300 and 900 mg/kg bw/day and HERA cited 260 and 780 mg/kg bw/day


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NOAEL of 85 mg/kg bw/day based on body weight gain, haematology, and clinical biochemistry.

Yoneyama et al. (1976) also conducted 9-month feeding studies in mice, in which groups of 8 or 9 male/females SLC-ICR mice were fed 0.6 or 1.8 % (500 and 1000 mg/kg bw/day) LAS in the diet or given drinking water containing 0.07, 0.2 or 0.6 % LAS for 9 months (Yoneyama et al., 1976 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013). These doses in drinking water corresponded to 100, 250, 600/900 mg/kg bw/day for males/females (HERA 2013; US EPA 2013) or 0, 133, 380 or 1140 mg/kg bw/day (REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020).

In the feeding study (Yoneyama et al., 1976, an increase in liver weight was seen in male and female mice at 500 mg/kg bw/day, with a significant decrease in lactate dehydrogenase and acid phosphatase in the liver and kidneys in male mice, respectively. A LOAEL of 500 mg/kg bw/day was determined by HERA 2013 and US EPA 2013. No point of departure was determined in the REACH dossier.

In the drinking water study (Yoneyama et al., 1976, a decrease in body weight was seen at the highest dose in males and females, as well as an increase in liver weight in females and significant decreases in renal Na, K ATPase leading to a NOAEL of 250 mg/kg bw/day (mid dose) being determined by HERA 2013 and US EPA 2013 based such effects. In contrast, the REACH dossier cited a NOAEL of 133 mg/kg bw/day (low dose), based on changes in body and organ weight and liver/kidney enzymes.

The Linear Alkylbenzene Alkylate Bottoms Consortium carried out a standard repeated dose test by administering 250, 500 or 1000 mg/kg bw/day to male and female rats (no further information available). Increased hyperplasia and hypertrophy of the follicular epithelial cells of the thyroid were observed at all doses.

Section G2.3.2 Irritancy and corrosivity

No data were identified relating to the irritancy potential of LAS in humans.

A number of skin and eye irritation studies have been carried out. In a study conducted according to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) 0.5 ml aqueous solution of 47 % LAS was administered to intact skin of three New Zealand rabbits under a semi-occlusive dressing for four hours (Huntingdon, 1986 cited in HERA 2004; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013; OECD SIDS 2005). Skin reactions were graded after 30 minutes following removal of the dressing and daily thereafter for 14 days. Irritation was reported in all animals 30 minutes after removal of the dressing (maximum score of 2). Symptoms worsened, and desquamation, necrosis, and hyperkeratinisation were noted by day 4. Symptoms were reversible in one animal by day 12, but not reversible in the other two animals. Overall, it was concluded that LAS is a skin irritant.

LAS (1, 2.5 and 5 %) was also administered to intact and abraded skin of six rabbits for 24 hours. 1 % and 2.5 % LAS did not cause any irritation whereas 5 % caused moderate irritation (BIOLAB, 1983, 1988 and 1989 cited in HERA 2013; US EPA 2013; OECD SIDS 2005).


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0.1 ml of an aqueous solution of 47 % LAS was administered to each eye of three rabbits (Huntingdon, 1986 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013; OECD SIDS 2005). In the first group the eyes were not rinsed, resulting in severe irritation which was not reversible in one rabbit after 21 days. In the second group, eyes were rinsed after four seconds and mild irritation was observed, which was fully reversible within seven days. In the third group, eyes were rinsed after 30 seconds and mild irritation was observed that was reversible within 14 days. Results for the unrinsed eyes were used for classification, due to OECD 405 protocol requiring at least 24 h exposure (HERA 2013).

Other studies were carried out in which 1, 5 or 50 % LAS was administered to six rabbits. LAS was not irritating at 1 %, moderately irritating at 5 %, and significantly irritating to the iris and conjunctivae at 50 %, persisting to day six (BIOLAB, 1984, 1988, 1989 cited in HERA 2013; US EPA 2013).

Lower concentrations (0.01 – 0.1 %) of LAS were also tested. 0.01 % did not cause any irritation in animals (species not reported). Slight irritation of the conjunctivae was seen at 0.05 %, considerable irritation at 0.1% within 2 hours, which resolved at 24 hours, and severe irritation at 0.5 % LAS for 24 h (severe irritation and oedema, increased secretion, turbidity of the cornea and disappearance of the corneal reflex). All effects were reversible within 120 hours (Imori et al., 1972 and Oba et al., 1968 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013).

Section G2.3.3 Sensitisation

Two Human Repeat Insult Patch Tests (HRIPT) were identified in humans. During the induction phase, 95 volunteers were treated with 0.10 % w/v LAS on upper arms under occlusive conditions for 24 h, three times per week for three weeks (Procter & Gamble, 1997 cited in HERA 2013; US EPA 2013; OECD SIDS 2005). After 14 to 17 days, volunteers underwent a challenge applied to the original and alternate arm for 24 h. No signs of sensitisation were seen.

2294 volunteers were also exposed to LAS as a raw substance and 17887 were exposed to LAS in formulations. No sensitisation reactions were reported (Nusair et al., 1988 cited in HERA 2013).

An occlusive epicutaneous test was also carried out (Matthies, 1989 cited in HERA 2013) in which 1 % LAS was applied once to volunteers. No reactions were seen.

LAS was negative in an OECD 406 study (Skin Sensitisation) (unnamed study, 1985 cited in REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020). Ten male and female guinea pigs underwent an induction with 0.1 ml Freund Complete Adjuvant (FCA), 0.1 ml of 25 % LAS in water, and 0.1 ml test substance in FCA in water (final concentration 25 %). On day 7, a second, epicutaneous challenge was carried out with 0.5 ml LAS (25 %) placed on gauze and then on the animals for 48 h. The animals were challenged on day 21 with 0.2 ml of 12.5 % LAS for 24 h and observations made after 48 and 72 h. No positive reactions were seen and the chemical is therefore non-sensitising.


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Guinea pigs injected intradermally with 1 % aqueous solution of LAS and topically challenged showed no skin sensitisation reactions (Shell Research Ltd, unpublished data cited in Soap and Detergent Association 1991).

A Magnusson and Kligman maximisation test in guinea pigs was also carried out. Animals received an induction dose of 0.5-5 % LAS via injection followed by a challenge with 1 % LAS. 76 % of animals tested positive for sensitisation effects. When the study was carried out again using the Buehler guinea pig test, LAS was negative to moderately positive with an induction dose of 2 % or greater. No sensitisation was observed at doses of 1.4 % LAS or less (Robinson et al., (date unknown cited in Soap and Detergent Association 1991; Little, 1991 cited in US EPA 2013; OECD SIDS 2005). The authors suggested that LAS was a weak sensitiser under exaggerated conditions.

Section G2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of LAS in humans.

LAS was negative in two Ames tests carried out according to OECD Guidelines (Huls, 1993; Schoeberl, 1993 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013; OECD SIDS 2005), in which 8-5000 µg/plate LAS (91.3 %) was tested with Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 with and without metabolic activation.

Other Ames tests were also negative, although they deviated from the OECD protocol as they used limited Salmonella typhimurium strains, did not report cytotoxicity or control data, and used low concentrations of LAS (Inoue et al., 1979 cited in HERA 2013; US EPA 2013; OECD SIDS 2005).

LAS (22.2 %) was negative in a transformation test with Syrian hamster embryo (SHE) cells without metabolic activation. Concentrations up to 50 µg/plate were tested (Inoue et al., 1980 cited in HERA 2013; Soap and Detergent Association 1991; OECD SIDS 2005).

0, 6, 10, 18, 30 and 60 µg/ml LAS and 0, 0.6, 1, 1.8, 3 and 6 µg/ml LAS was tested in CHO cells HPRT assay with and without metabolic activation, respectively, according to OECD guideline 476 (In Vitro Mammalian Cell Gene Mutation Test). LAS was cytotoxic above 50 and 100 µg/ml with and without metabolic activation, respectively. There was no biologically significant increase in mutation frequency in treated cells; hence it was concluded that LAS was not mutagenic (Anon, 1995 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020)).

In a chromosomal aberration assay, carried out in accordance with OECD 473 (In Vitro Mammalian Chromosome Aberration Test), CHO cells were exposed to 2.5, 5, 10, 15, 20, 26, 33, and 39 µg/ml LAS with S9, and 20, 39, 58, 78, 104, 130, and 156 µg/ml without S9. LAS produced equivocal results in cells with metabolic activation but negative without S9 mix. The authors concluded that LAS is weakly clastogenic at cytotoxic concentrations but negative at concentrations where cytotoxicity was not seen (Murie and Innes, 1997 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020)).


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An in vivo chromosomal aberration assay in male mice was also carried out (Inoue et al., 1979 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013). LAS (0, 200, 400 or 800 mg/kg bw/day) was administered for one and five days via oral gavage. There was no significant difference between dose groups and controls in the number of chromosomal aberrations in bone marrow 6, 24 or 48 hours after administration.

In a further cytogenetic assay (Masabuchi et al., 1976 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005), male rats and mice were administered 0.9 % LAS (450 mg/kg bw/day and 1170 mg/kg bw/day, respectively) for 9 months via their feed. No increase in chromosomal aberrations were observed in treated groups compared with controls.

In a micronucleus assay, male mice received a single i.p. injection of 100 mg/kg bw/day LAS. No differences in the incidences of polychromatic erythrocytes with micronuclei in the bone marrow in the treated group were observed compared to the control group (Kishi et al., 1984 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005).

Masabuchi carried out a dominant lethal assay in male mice (Masabuchi et al., 1976 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013). LAS (0.6 %; 300 mg/kg bw/day) was administered to 7 male mice in the feed for 9 months. One male mouse was mated with two untreated female mice. There were no significant differences in fertility, mortality of ova and embryos, number of surviving fetuses, or the index of dominant lethal induction between the treated animals and controls.

Section G2.3.5 Carcinogenicity

No data were identified relating to the carcinogenicity of LAS in humans.

Four carcinogenicity studies are available. LAS (0.02, 0.1 and 0.5 %; 10, 50 or 250 mg/kg bw/day) was administered to male and female rats (50/dose/sex) for two years in the diet. There was no difference in the incidence of tumours in treated and control groups (Buehler et al., 1971 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005).

In another study, Wistar rats were exposed to 0.01, 0.05 or 0.1 % LAS (34.55 %) in drinking water for two years, corresponding to 20, 100 or 200 mg/kg bw/day (Tiba et al., 1972 cited in HERA 2013; US EPA 2013). No treatment related changes in growth, mortality, organ weight or histopathology were reported. There was no description of tumours in the report (HERA 2013; OECD SIDS 2005).

Male and female Wistar rats (5-15/dose/sex) were also given LAS 0.04, 0.16. 0.6 %; 20, 80 and 300 mg/kg bw/day) in the feed for 1, 3, 6, 24 months (Fujii. et al.,1977; Yoneyama et al., 1977 cited in HERA 2013). The authors concluded that LAS did not have any adverse effects on rats.

Lastly, male and female rats (62/group) were administered 0.1 % LAS (200 mg/kg bw/day) for 26 months in drinking water. No treatment-related tumours were reported (Endo et al., 1980 cited in HERA 2013).


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Section G2.3.6 Reproductive and developmental toxicity

No data were identified relating to the reproductive and/or developmental toxicity of LAS in humans.

Several reproductive toxicity studies have been carried out, including two three-generation and a four-generation study. Male and female Charles River rats (50/dose) were treated with 0.02, 0.1 and 0.5 % LAS (14, 70 and 350 mg/kg bw/day) in the diet in the first three-generation reproductive toxicity study (Buehler et al., 1971 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). The F0 generation was administered LAS for 84 days prior to mating. The F1a generation was sacrificed at 21 days of age. Ten days later, females were re-mated with different males from the same group. From this F1b generation, 20 males and females were selected for mating and were fed LAS-containing diets for 80-85 days until mating, to produce the F2b generation. The F2b generation was further treated for eight weeks prior to mating. General reproduction, including fertility gestation, parturition, neonatal viability, lactation, and post-weaning growth, was not affected in any of the test groups and did not deviate from controls. The NOAEL parental, NOAEL F1 offspring and NOAEL f2 offspring were all 350 mg/kg bw/day (highest dose tested).

In a second three-generation reproductive toxicity study, rats (10-20 dose/sex) were administered liquid detergent containing 17 % LAS and 7 % alkyl ethoxylate sulphate via feed (Palmer et al., 1974 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). Dietary concentrations of 0, 0.08, 0.4 or 2 % (0, 40, 200 or 1000 mg/kg bw/day) of the formulation were continuously administered throughout three generations for 60 days prior to mating. The corresponding concentrations of LAS were 0, 6.8, 34 or 170 mg/kg bw/day. No effect on mating performance, pregnancy rate, duration of pregnancy or incidence of malformations was reported. A NOAEL of 170 mg/kg bw/day was determined (highest dose tested).

In the four-generation reproductive toxicity study, male and female Wistar rats were administered 0.1 % LAS (70 mg/kg bw/day) in drinking water. No effects on fertility, parturition, gestation period, or lactation in any of the generations were reported. No further information available (Endo et al., 1980 cited in HERA 2013).

Palmer and colleagues also carried out a number of developmental studies on rats, mice and rabbits (Palmer et al., 1975). Charles River rats (20/dose) were administered LAS at doses of 0.2, 2, 300, 600 mg/kg bw/day on day 6-15 of pregnancy by gavage and sacrificed at day 20 of gestation (Palmer et al.,1975 cited in HERA 2013; unnamed report, 1971 cited in REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). A decrease in body weight gain, associated with disturbance of the gastrointestinal tract, was reported in the top dose group. The decrease partially recovered toward the end of the dosing period. Pregnancy rates were comparable at all doses, and no differences were observed among the dose groups and controls with respect to number of litters, viable young, litter weight, fetal weight, embryonic deaths, implantations, corpora lutea, pre- and post-implantation embryonic loss, major malformations, minor visceral or skeletal anomalies, or incidence of pups with extra ribs. In the HERA document a maternal NOAEL of 300 mg/kg bw/day and a developmental NOAEL of


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600 mg/kg bw/day were determined. However, the REACH dossier cites a NOAEL of 300 mg/kg bw/day for both maternal and developmental toxicity.

In the mouse study, CD-1 mice (20/dose) were administered 0.2, 2, 300, 600 mg/kg bw/day LAS by gavage on days 6-15 of pregnancy, then sacrificed on day 17 (Palmer et al.,1975 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). Mortality was seen in parent animals at 300 and 600 mg/kg bw/day. At 300 mg/kg bw/day, weight gain was decreased only for the first four days. At doses with no maternal toxicity, no differences were observed among the dose group and controls with respect to number of litters, viable young, litter weight, fetal weight, embryonic deaths, implantations or post-implantation embryonic loss. At maternally toxic doses, there was increased fetal loss and reduced litter size due to total litter loss, which was considered to be a secondary effect of maternal toxicity. Authors of the HERA document stated that no NOAEL could be determined due to the large difference between the NOAEL in maternal animals (2 mg/kg bw/day) and the LOAEL (300 mg/kg bw/day). A developmental NOAEL of 300 mg/kg bw/day was determined as no assessment was possible at 600 mg/kg bw/day due to the high mortality rate of maternal animals. In the REACH dossier a maternal NOAEL of 2 mg/kg bw/day and a developmental NOAEL of 300 mg/kg bw/day were determined.

Lastly, New Zealand White rabbits (13/dose) were administered 0.2, 2, 300, 600 mg/kg bw LAS by gavage at days 6-18 of pregnancy and sacrificed at day 29 (Palmer et al.,1975 cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). No effect on bodyweight changes and pregnancy rates of parent animals was seen at 0.2 and 2 mg/kg bw/day. At 300 and 600 mg/kg bw/day, parent animals showed anorexia, diarrhoea, weight loss and death. No effects on litter parameters were seen at lower doses. Maternal toxicity precluded assessment of the effects on litter parameters at higher doses. The HERA document again concluded that reliable NOAELs could not be determined due to the large difference between the maternal dose that caused no effects (2 mg/kg bw/day) and the maternal LOAEL (300 mg/kg bw/day). In addition, effects on litter parameters could not be determined at 300 mg/kg bw/day due to the high mortality rate in maternal animals. However, in the REACH dossier a NOAEL of 2 mg/kg bw/day for maternal and developmental toxicity was determined.

Tiba et al. (1976) also carried out a developmental study in pregnant female rats (16/dose), which were administered 0.1 or 1.0 % LAS (80 and 780 mg/kg bw/day) in the diet from day 0 to 20 of gestation (Tiba et al.,1976 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013). At the highest dose, the number of offspring and the weaning rate were low compared to controls. No other adverse effects were reported. A maternal and developmental NOAEL of 780 mg/kg bw/day was determined.

In another developmental study, female rats (40/dose) and rabbits (22/dose) were administered 0.1 % LAS (383 mg/kg bw/day for rats and 3030 mg/kg bw/day for rabbits) in drinking water from day 6 to 15 (rats) and day 6 to 18 of pregnancy (rabbits) (Endo et al.,1980 cited in HERA 2013; US EPA 2013). A slight decrease in body weight gain was seen in rabbits. The litter parameters of both species did not show any significant differences from those of the controls. Delayed ossification was observed in rabbits, but there was no increase in malformations in either the rabbits or the rats. A maternal and developmental NOAEL of 383


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mg/kg bw/day was determined in rats and maternal and developmental LOAEL of 3030 mg/kg bw/day in rabbits.

Doses of 40, 400 mg/kg bw/day LAS were administered daily by oral gavage to female mice (13-14/dose) from day 0 to day 6 or day 7 to 13 of pregnancy (Takahashi et al.,1975 cited in HERA 2013; US EPA 2013; OECD SIDS 2005). In mice given 400 mg/kg bw/day, pregnancy rate was decreased in the top dose group compared with controls but there was no increase in malformations. No information was available on maternal toxicity but authors stated it appeared likely that maternal toxicity occurred at the top dose (HERA 2013). A maternal NOAEL of 40 mg/kg bw/day and a developmental NOAEL of 400 mg/kg bw/day was determined.

LAS (10, 100, 300 mg/kg bw/day) was also administered to female mice (25-33/dose) daily by gavage on day 6 to 15 of pregnancy (Shiobara et al.,1976 cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005). A decrease in body weight was seen in dams at all doses. At the highest dose, two dams died, and one had a premature delivery and all fetuses died. In the living fetuses, decreased body weight and delayed ossification were reported, although no malformations were observed. A maternal LOAEL of 10 mg/kg bw/day and a developmental NOAEL of 300 mg/kg bw/day was determined.

Section G2.3.7 Specific considerations

No data were identified relating to the other endpoints of toxicity of LAS.

Section G2.3.8 Summary of human relevant health effects

• No data were identified relating to the toxicokinetics of LAS in humans.

• In animals LAS is excreted primarily via urine and to a lesser extent via bile.

• LAS is of moderate acute oral and dermal toxicity.

• LAS is moderately irritating to skin, and severely irritating to eyes.

• No sensitisation potential was seen with LAS either in animals or humans.

• Repeat dose oral studies showed effects of LAS in liver and kidneys.

• Mutagenicity and genotoxicity studies generally indicate LAS does not have mutagenic potential apart from it being weakly clastogenic at cytotoxic concentrations.

• There is no evidence that LAS has carcinogenic potential.

• Reproductive and developmental toxicity studies did not show direct adverse effects of LAS following oral exposure. Any results seen were due to maternal toxicity.

Section G3.0 Identification of relevant point(s) of


Section G3.1 Critical endpoints for human health risk assessment

purposes

The POD for use in the risk assessment (see Section G4.0) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to LAS through any exposure route. However, a number of oral repeated dose experimental studies have been


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carried out for LAS, for systemic and specific organ toxicity. These are summarised in Table G3.1.


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Table G3.1 Oral route toxicity studies for LAS

Type/Duration Species/Strain Dose range (mg/kg bw/day)/route of exposure



Rat / CRJ-SD 125-500 via gavage At all doses – soft stools in both sexes and decreases in body weight gain in both sexes and organ weight changes (liver in females and heart in males) At doses ≥250 mg/kg bw/day - changes in serum biochemistry and decreased kidney weight (females) and heart weight (males) NOAEL of 125 mg/kg bw/day derived based on clinical biochemistry and body weight changes.

Ito et al.,1978 (cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991)


Rat / CRJ-SD 50 and 250 via the diet No treatment related effects were seen. NOAEL of 50 mg/kg bw/day (highest dose tested).

Oser, 1965 (cited in HERA 2013)


Rat / strain unknown 8.8-200 via the diet No treatment related effects were seen. NOAEL of 200 mg/kg bw/day (highest dose tested).

Kay et al., 1965 (cited in HERA 2013)

Subchronic/ 26 weeks

Mouse / strain unknown 20 via drinking water At 20 mg/kg bw/day – hepatotoxicity at one and six months. LOAEL of 20 mg/kg bw/day (only dose tested).

Watari et al., 1977 (cited in HERA 2013; Soap and Detergent Association 1991)

Subchronic/ 26 weeks

Rat / Wistar SLC 40 - 1030 via the diet At doses ≥115 mg/kg bw/day - changes in haematological and biochemistry parameters, increased cecum weight and decreased kidney weight NOAEL of 400 mg/kg bw/day based on haematology, biochemistry and organ weight changes.

Yoneyama et al.,1972 (cited in HERA 2104; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005)

Subchronic/ 9 month

Rat / Wistar JCR 260/300/500 – 780/900/1000 via the

Feeding study. Yoneyama et al.,1976 (cited in HERA 2013;


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diet or 85-430 via drinking water

At the low dose – an increase in liver weight (% not given) and changes in biochemical parameters were seen Drinking water study At the mid dose - decrease in lactate dehydrogenase and Na, K-ATPase, an increase in alanine transferase and a significant decrease in body weight gain NOAEL of 85 mg/kg bw/day (drinking water study) was derived based on clinical biochemistry and organ weight ratios

REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013; OECD SIDS 2005)

Subchronic/ 9 month

Mouse / SLC-ICR 500-1000 via the diet or 100 – 600/900 or 133 – 1140 via drinking water

Feeding study. At the low dose – increased liver weight in both sexes and a decrease in in lactate dehydrogenase and acid phosphatase in the liver and kidneys in male mice Drinking water study At the low dose – increase in relative liver weight and alteration of liver enzymes LOAEL of 500 mg/kg bw/day (feeding study) and NOAEL of 133 mg/kg bw/day (drinking water study) were derived based on increased liver weight and clinical chemistry parameters.

Yoneyama et al.,1976 (cited in HERA 2013; REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; US EPA 2013)

Three generation reproductive toxicity study / F0 generation 84 days prior to mating; F1b generation 80-85 days prior to mating; F2

Rat / Charles River 14 – 350 via the diet No treatment related effects were seen. NOAEL of 350 mg/kg bw/day (highest dose tested).

Buehler et al., 1971 (cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005)


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generation 8 weeks prior to mating

Three generation reproductive toxicity study / 60 days prior to mating

Rat / strain unknown 6.8 - 170 via the diet No treatment related effects were seen. NOAEL of 170 mg/kg bw/day (highest dose tested).

Palmer et al., 1974 (cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005)

Four generation reproductive toxicity study

Rat / Wistar 70 via oral drinking water

No treatment related effects were seen. No NOAEL is derived as only one dose was used.

Endo et al., 1980 (cited in HERA 2013)

Development toxicity study / GD 6 - 15

Rat / Charles River 0.2 – 600 via oral gavage At 600 mg/kg bw/day – decrease in body weight gain that was partially recovered towards the end of the study. No effects seen in the fetus. NOAEL of 300 mg/kg bw/day was derived for maternal toxicity and a NOAEL of 600 mg/kg bw/day for developmental toxicity.

Palmer et al.,1975 (cited in HERA 2013; unnamed report, 1971 cited in REACH registration dossier for Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts 2020; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005)


Mouse / CD1 0.2 – 600 / via oral gavage

At 300 mg/kg bw/day – decreased body weight gain seen for the first four days. At 300 and 600 mg/kg bw/day – mortality observed and increased fetal loss and reduced litter size due to total litter loss, which was a secondary effect due to the maternal toxicity. NOAEL of 2 mg/kg bw/day was derived for maternal toxicity and a NOAEL of 300 mg/kg bw/day for developmental toxicity.



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Rabbit / New Zealand White

0.2 – 600 / via oral gavage

At 300 mg/kg bw/day – anorexia, diarrhoea, weight loss and death occurred. At 0.2 and 2 mg/kg bw/day - no effects on litter parameters seen. Effects at higher doses could not be assessed due to maternal toxicity. NOAEL of 2 mg/kg bw/day was derived for maternal toxicity and developmental toxicity.


Developmental study / GD 0 – 20

Rat / strain unknown

80 – 780 mg/kg bw/day via the diet

At 780 mg/kg bw/day - the number of offspring and the weaning rate was low compared to controls. NOAEL of 780 mg/kg bw/day was derived for maternal toxicity and developmental toxicity.

Tiba et al.,1976 (cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013)

Developmental study / GD 6 – 15 (rat) and 6 – 18 (rabbit)

Rat / strain unknown Rabbit / strain unknown

383 (rat) and 3030 (rabbit) via drinking water

At 383 mg/kg bw/day (rat) – no effects were seen. At 3080 mg/kg bw/day (rabbit) slight decrease in body weight and delayed ossification. NOAEL of 383 mg/kg bw/day was derived for maternal toxicity and developmental toxicity (rat) and a LOAEL of 3080 mg/kg bw/day was derived for maternal toxicity and developmental toxicity (rabbit) (only doses tested).

Endo et al.,1980 (cited in HERA 2013; US EPA 2013)

Developmental study / GD 0 – 6 or GD 7 - 13

Mouse / strain unknown 40 - 400 mg/kg bw/day via oral gavage

At 400 mg/kg bw/day – pregnancy rate, but not malformations, was reduced compared with controls. NOAEL of 40 mg/kg bw/day was derived for maternal toxicity and a NOAEL of 400 for developmental toxicity.

Takahashi et al.,1975 (cited in HERA 2013; US EPA 2013; OECD SIDS 2005)

Developmental study / GD 6 - 15

Mouse / strain unknown 10 – 300 mg/kg bw/day via oral gavage

>10 mg/kg bw/day – decrease in bodyweight in dams and delayed ossification, but not malformations. NOAEL of 10 mg/kg bw/day was derived for maternal toxicity and a NOAEL of 300 for developmental toxicity.

Shiobara et al.,1976 (cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005)


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Carcinogenicity study / 2-year

Rat / strain unknown 10 – 250 mg/kg bw/day via the diet

No treatment-related effects were seen.

Buehler et al., 1971 (cited in HERA 2013; Soap and Detergent Association 1991; US EPA 2013; OECD SIDS 2005)


Rat, Wistar 20 - 200 mg/kg bw/day in drinking water


Tiba et al., 1972 (cited in HERA 2013; US EPA 2013)


Rat, Wistar 10 – 300 mg/kg bw/day via the diet


Fujii. et al.,1977; Yoneyama et al.,1977 (cited in HERA 2013)

Carcinogenicity study / 26-months

Rat (strain unknown) 200 mg/kg bw/day in drinking water


Emdo et al., 1980 (cited in HERA 2013)



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The collated findings indicate that the most sensitive effects following oral exposure to LAS include body weight and organ weight changes, haematological effects and changes in clinical chemistry (discussed further in section G3.3). There is no evidence from currently available studies that LAS exposure is linked to carcinogenicity or reproductive or developmental toxicity.

Section G3.2 Current health-based guidance values


Section G 3.3 Identification of POD to be used for the risk

assessment

The lowest reported NOAEL was 2 mg/kg bw/day in developmental toxicity studies in mice and rabbits, based on maternal toxicity (anorexia, diarrhoea, weight loss and death) seen at 300 mg/kg bw/day (Palmer et al., 1975). However, due to the large dose spacing between 300 and 2 mg/kg bw/day, some authoritative bodies have deemed it inappropriate to determine a NOAEL from this study.

Maternal NOAELs of 10 and 40 mg/kg bw/day were determined in two other developmental toxicity studies (Takahashi et al.,1975; Shiobara et al.,1976). However, both have been discounted, as in one study no maternal toxicity data were reported and in the other only a decrease in body weight of dams was seen, with no additional information provided.

A NOAEL of 85 mg/kg bw/day was determined by Yoneyama et al., (1976) in a 9-month drinking water study in rats, based on body weight gain, haematology, clinical biochemistry and organ weight ratios. This is deemed the most reliable POD based on sensitive endpoints.

Section G4.0 Drinking water risk assessment


The most sensitive endpoint is changes in a decrease in body weight gain, haematology and clinical biochemistry, observed in a 90-day feeding study in rats.


Based on the data obtained, the NOAEL is considered to be 85 mg/kg bw/day, based on changes in liver and kidney enzymes. This is used as the POD.

Assessment Factors (AF)



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AF Justification




1 NOAEL used as POD


Inadequate databases 1 No long term studies are available

Total AF 300








Exposure assessment

The maximum concentration of LAS measured in drinking water, as reported by WRc (2014), was 37.0 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.


• 1.23 µg/kg bw/day for a 60 kg adult drinking 2 L water per day

• 3.70 µg/kg bw/day for a 10 kg child drinking 1 L water per day

• 5.55 µg/kg bw/day for a 5 kg infant drinking 0.75 L water per day


The maximum intake of LAS via drinking water by adults, children and infants is less than the TDI (HQ <1) and the measured concentration of LAS in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to LAS via drinking water.

Risk communication

The MOEs for LAS, based on the NOAEL of 85 mg/kg bw/day and maximum intakes of 123, 370 and 5.55 µg/kg bw/day are 68919, 22973 and 15315 for adults, children and infants, respectively. The MOEs indicate that exposures are not of concern in terms of risk to public health.


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References

ECHA REACH registration dossier (2020). Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts. https://echa.europa.eu/registration-dossier/-/registered-dossier/15879 Accessed August 2020.

HERA (2013). Human & Environmental Risk Assessment on ingredients of European household cleaning products; LAS Linear Alkylbenzene Sulphonate (CAS 68411-30-3). Accessed Jan 2020.

OECD SIDS (2005). SIDS initial assessment report for 20th SIAM. Linear Alkylbenzene Sulfonate (LAS). Accessed Jan 2020.

Soap and Detergent Association (1991). Environmental and Human Safety of Major Surfactants. Volume I. Anionic Surfactants. Part 1. Linear Alkylbenzene Sulfonates. Accessed Jan 2020.

US EPA (2013). Alkylbenzene Sulfonates. Final Work Plan. Accessed Jan 2020.




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Section H: Cocamidopropyl betaine

(CAPB)

Section H1.0 Chemical identification, use and


Section H1.1 Reasons for consideration

A report by WRc published in 2014 assessed the impact on drinking water of the use of the amphoteric surfactant cocamidopropyl betaine (CAPB) as a foam boosting agent in shampoo, hand soap and cosmetics. Consumption of the surfactant in the UK increased from 4604 tonnes/year in 2007 to 6863 tonnes/year in 2012. Although current legislation8 is in place to limit the use of nonbiodegradable surfactants, total degradation of permitted surfactants cannot be guaranteed. With the high volume of use of CAPB there is therefore a possibility of the surfactant reaching drinking water (WRc, 2014).

Levels of CAPB in drinking water were estimated as a maximum of 145 µg/L. The authors noted that this was based on likely overestimated initial concentrations entering the wastewater treatment works. CAPB is included for further consideration here as the estimated exposures through drinking water and bathing comprised a significant proportion (45 %) of the intended use (SED of 0.01 mg/kg bw/day), meaning that the potential for significant exposure in the general population was high (WRc, 2014).

Section H1.2 Identification and physicochemical properties

CAPB is an amphoteric surfactant which is zwitterionic in nature, i.e. both anionic and cationic structures are found in one molecule. As a betaine, the cationic form of CAPB is present at very low pH, but it does not have anionic properties under alkaline conditions (HERA, 2005). CAPB is produced from the hydrolysis of coconut oil, a natural product of mixed fatty acids which varies slightly in composition; the physicochemical properties of CAPB are therefore difficult to standardise and estimated data are outlined in Table H1.1.

CAPB is listed with Cas Nos. 61789-40-0, 86438-79-1 and 83138-08-3, referred to as ‘cosmetic grade’ by WRc (WRc, 2014). The main constituent of CAPB is lauramidopropyl betaine (present at around 50 %) which has the Cas No. 4292-10-8, also referred to as ‘technical grade’ (WRc, 2014).

8 Regulation (EC) No 648/2004 of the European Parliament and of the Council of 31 March 2004 on detergents.


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Table H1.1 CAPB identification and physicochemical properties


Cas No

61789-40-0

IUPAC name 1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-coco acyl derivs., hydroxides, inner salts

Chemical Group Alkylamidopropyl betaines

Physical form Solid

DSL name 1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-coco acyl derivs., hydroxides, inner salts

Common name Cocamidopropyl betaine

Formula C19H38N2O3

Molecular weight 342 (typical for C12)

Structure

Solubility Insoluble

Melting point >25 °C (solid)

Boiling point >300 °C (estimated)9

Density 0.32 (estimated)

Log Pow Not applicable due to dispersibility

Vapour Press. <1.0×10-10 mm Hg at 25°C (estimated)2

Sources: WRc 2014; PubChem; ChemID Plus; ECHA REACH registration dossier for CAPB

Section H1.3 Hazard classifications

CAPB has been assigned a hazard category of Skin Irrit. 2 with hazard statement H315 – “causes skin irritation”, Eye Irrit. 2 with hazard statement H319 – “causes serious eye irritation” and Skin Sens. 1 with hazard statement H317 – “may cause an allergic skin reaction” in the REACH dissemination page (ECHA REACH registration dossier for Cocamidopropyl betaine, 2020).

The EU notified classification and labelling according to CLP regulation (EC) No. 1272/2008, includes Skin Irrit. 2 with hazard statement H315 – causes skin irritation, Eye Irrit. 2 with hazard statement H319 – causes serious eye irritation, Eye damage 1 – with hazard statement H318 - causes serious eye damage and Skin Sens. 1 with hazard statement H317 – may cause an allergic skin reaction. There is no harmonised classification.

9 US EPA (2014)


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Section H1.4 Occurrence, production and use.

CAPB is used in a wide variety of PCPs including shampoos, bath products, cleansing agents, shower gels, bath foam, liquid soaps and skin care products. In addition, CAPB is also widely used in DCPs such as laundry detergents, hand dishwashing liquids and hard surface cleaners (HERA, 2005). CAPB functions in these products to reduce static, condition skin and hair, moderate viscosity and as a foaming agent. It is classed as a high production volume chemical by the US EPA (EPA, 2011).

The alkylamidopropyl betaine is produced in a two-step reaction involving dimethylaminopropylamine (DMAPA) and the fatty acids from coconut or palm kernel oil, which have lauric acid (or its methyl ester) as the main constituent. The second step constitutes reaction with chloroacetic acid, which forms a quaternary ammonium centre (HERA, 2005). Cocamidopropyl betaine is most commonly supplied as a 30 % w/w aqueous solution (a manufacturing concentrate) to a detailed manufacturer’s specification (APVMA, 2017). Around 59,000 metric tons of betaines were produced in 2002 in Western Europe, with around 50 % being used as a cosmetic ingredient (around 29,500 tons/year) and the remainder as a detergent. The Cosmetic Ingredient Review (CIR) Expert Panel reported use of CAPB in cosmetics at concentrations between 0.005 and 11 % (Burnett et al., 2012) whilst HERA reported CAPB use in cleaning products at concentrations between 0.1 and 30 % (HERA, 2005).

Section H1.5 Human exposure

Occupational exposure is a possible source of exposure to CAPB in humans, with the dermal and inhalation routes being key. Consumer exposure is likely to be greater through occupational (e.g. hairdressers, cleaners) and non-occupational uses of products containing CAPB. Several scenarios have been identified including both direct and non-direct dermal exposure, inhalation of aerosols from cleaning sprays, and oral ingestion resulting from residues deposited on dishes and/or ingestion of drinking water that has been contaminated with CAPB. Estimated intakes for indirect exposure via all routes of 0.58 µg/kg bw/day and 0.99 µg/kg bw/day respectively were reported (HERA, 2005).

Section H2.0 Human relevant health effects

Section H2.1 Toxicokinetics

No data were identified relating to the toxicokinetics of CAPB in humans or experimental animals following exposure via any route.

An experimental toxicokinetics study has been reported for the major fraction (around 50 %) of CAPB, lauramidopropyl betaine (LB), which has previously been used to provide read-across data for CAPB (HERA, 2005). 14C-labelled LB was administered by oral gavage at doses of 0, 10 and 30 mg/kg bw to male and female Wistar rats which were followed for 48 h after dosing. Absorption of LB from the GI tract at doses of 10 and 30 mg/kg bw was poor, with only 5 % of the total dose being excreted in urine and < 2 % in exhaled air, with 1 % remaining in the carcass after 48 h. The remainder of the dose was excreted in faeces as unchanged parent material. Distribution of LB following absorption was to the organs associated with


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urinary excretion, i.e. liver, kidney cortex and urinary bladder (Unilever Research, 1992 - cited in HERA, 2005).

Section H2.2 Acute toxicity

No data were identified relating to the acute toxicity of CAPB in humans via any route of exposure.

The acute oral toxicity of CAPB has been tested in several rat strains (Wistar, Sprague-Dawley and CD), administered by oral gavage to males and females as an undiluted solution (containing 30 – 35.5 % active solution). Studies pre-dated OECD Guidelines and as these were industry studies, protocols are unavailable for evaluation. An oral LD50 of ≥ 4900 mg/kg bw has been estimated from these studies (HERA, 2005). Sub-lethal effects included diarrhoea, nasal haemorrhage, salivation, decreased motor activity, coordination disturbance and abnormal body posture.

Section H2.3 Repeat dose toxicity

Section H2.3.1 Systemic effects

No data was identified relating to the systemic effects of CAPB in humans via any route of exposure.

In a 28-day study carried out to OECD guideline 407 (Repeated Dose 28-day Oral Toxicity Study in Rodents), male and female Sprague-Dawley rats were administered CAPB as a 30 % active solution by oral gavage at doses of 0, 250, 500 and 1000 mg/kg bw/day for 5 days per week (Henkel KGaA, 1991 cited in HERA, 2005 and Burnett et al., 2012). Two recovery groups were included which were assessed 28 days following cessation of exposure. Oedema of the forestomach was evident at necropsy in females at the highest dose, and in both males and females at this dose following histopathological investigation. Histopathology also revealed the presence of acanthosis and oedema of the forestomach mucosa and multiple ulcerations and hyperplasia of the forestomach epithelium, with more severe findings in females. The authors noted that the effects in the forestomach were reversible and were likely due to the irritant properties of CAPB. No further macroscopic or microscopic effects were noted in other organs, nor were any changes seen in clinical chemistry or haematological parameters. NOAELs of 500 and 1000 mg/kg bw/day were derived with respect to forestomach and systemic effects respectively (Henkel KGaA, 1991 cited in HERA, 2005 and Burnett et al., 2012).

A 90-day study carried out to OECD guideline 408 (Repeated Dose 90-day Oral Toxicity Study in Rodents) has also been described for CAPB (Th. Goldschmidt AG, 1991c cited in HERA, 2005 and Burnett et al., 2012). Male and female Sprague-Dawley rats were administered a 30 % active solution of CAPB at doses of 0, 250, 500 and 1000 mg/kg bw/day for 5 days per week via oral gavage. Gross pathology showed ulceration of the stomach at the fundus and cardia regions at the highest dose, with forestomach gastritis evident at both the mid and high doses. Histopathological analysis confirmed the presence of squamous hyperplasia, submucosal oedema and inflammatory cell infiltration in males and females. No further macroscopic or microscopic effects were noted in other organs, nor were any changes seen in clinical chemistry or haematological parameters. HERA proposed a NOAEL of 1000 mg/kg


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bw/day with respect to systemic effects and of 250 mg/kg bw/day based on forestomach findings (Th. Goldschmidt AG, 1991c – cited in HERA, 2005 and Burnett et al., 2012).

Section H2.3.2 Irritancy and corrosivity

Vilaplana et al. (1992) reported slight skin irritation in humans exposed to CAPB at concentrations between 0.15 and 3 % (w/v) under occlusion for 2 days. The authors also reported a dose-dependent increase in irritation with increasing levels of the impurity amidoamine and concluded that this was most likely to be the cause of the irritation seen. Slight skin irritation was also reported in some (15 of 1200; 1.25 %) patients with eczema tested with a 1 % (w/v) solution of CAPB under occlusion for 2 days (Angelini et al., 1995).

GHS criteria were applied to findings reported by Beardesca et al. (1990) in which the irritant properties of a 7 % active solution of CAPB to humans was assessed using the plastic occlusion stress test (POST) technique. Following occlusion, a statistically significant increase above controls was found for skin surface water loss in the presence of CAPB, allowing classification of CAPB as a skin irritant in humans.

In a study carried out to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) CAPB applied as an 80 % active paste under semi-occlusive conditions for 4 h did not cause erythema or oedema of rabbit skin. Similarly, under the same conditions, a 30 % active solution of CAPB showed minimal skin irritation. There have been reports of irritating properties with 25 % and 30 % active solutions under more stringent occlusion conditions, but this is a deviation from OECD Guideline 404. Other skin irritation studies in rabbits have been carried out according to the Draize protocol, with CAPB being applied to intact and abraded skin under occlusive conditions for 24 h. Mild irritation was reported for a 10 % active solution and highly irritating/corrosive outcomes for solutions of 14 and 15 % CAPB (HERA, 2005).

Several acute irritation studies in the rabbit have been reported. One study carried out to OECD guideline 405 (Acute Eye Irritation / Corrosion) showed irreversible irritation by CAPBD at a concentration of 80 % active ingredient. All other available studies included deviations to the Guideline protocol and showed CAPB solutions with 25 – 30 % active ingredient to be irreversibly irritating or highly irritating, solutions of 14 – 15 % to be highly irritating, and levels ≤10 % to be mildly to moderately irritating with reversibility after 14 days. Inclusion of an eye rinse step after 30 sec exposure to a 15 % solution of CAPB did not affect the irritation score but did result in effects being reversible (HERA, 2005). CAPB meets the criteria for classification as an eye irritant under the CLP regulation.

Section H2.3.3 Sensitisation

The sensitising potential of CAPB has been assessed in three human volunteer studies using between 0.9 % and 10 % active solutions. No evidence of sensitisation was seen. Although in one study slight reactions were noted, these were concluded by the authors to be due to the irritant properties of CAPB, and have since been attributed to the presence of the sensitising impurities amidoamine (AA) and dimethylaminopropylamine (DMAPA) (HERA, 2005). The CIR expert panel reported that levels of AA and DMAPA varied between manufacturers, with levels between 0.3 to 3.0 % and 0.0003 to 0.02 % respectively (Burnett et al., 2012). A number of studies have also been carried out in individuals occupationally exposed to CAPB (for


122

example, hairdressers), or those with suspected contact dermatitis or unspecified eczema (Andersen et al., 1984, Cameli et al., 1991, Su et al., 1998, Korting et al., 1992, Van Haute et al., 1983, Taniguchi et al., 1992, Mowad, 2001, Ross et al., 1991, Bonneau et al., 1990 – cited by HERA, 2005). Positive reactions varied between 0.3 and 8.6 % which was considered very low when viewed in the context of the use of CAPB in a wide range of products (Jackson, 2001 – cited in HERA, 2005). The impurity AA undergoes hydrolysis on the skin surface releasing DMAPA and both substances have been shown to produce skin reactions in individuals with a CAPB allergy (Angelini et al., 1995; Fowler et al., 1997; McFadden et al., 2001 – cited by HERA, 2005). In addition, these substances have been reported to be key in causing skin sensitisation (Pigatto et al., 1995, Angelini et al., 1995, McFadden et al., 2001, Hunter et al., 1998, Fowler et al., 1997, Foti et al., 2003 – cited by HERA 2005). These studies highlight the importance of controlling the specification of CAPB.

The sensitising potential of CAPB has been assessed in two Magnusson and Kligman guinea pig maximisation tests (GPMT). Arimura et al. (1998) reported negative findings whilst Rantuccio et al. (1983) found CAPB to be weakly positive. However, no re-challenge was performed to verify the result. A Draize and modified Draize test performed with CAPB were also negative (HERA, 2005). CAPB meets the criteria for classification as a weak sensitiser under CLP regulation.

Section H2.3.4 Genotoxicity and mutagenicity

No data were identified relating to the genotoxicity or mutagenicity of CAPB in humans.

A mouse micronucleus assay was reported in which OF1 mice were administered between 20 and 200 mg/kg bw of a 27 % active CAPB solution by i.p. injection (Goldschmidt France, 1987 – cited in HERA, 2005). No increase in the incidence of micronucleus formation was seen compared to negative controls.

In vitro studies have also been carried out to assess the mutagenic potential of CAPB (HERA, 2005). When tested with Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 at doses between 1 and 5000 µg/plate carried out according to OECD guideline 471 (Bacterial Reverse Mutation Test), CAPB was negative in both the presence and absence of S9. Cytotoxicity was noted at doses ≥580 µg/plate, which is in line with its bactericidal properties (Henkel KGaA, 1988; Kao Corporation, 1996 – cited in HERA, 2005). CAPB was negative in mouse lymphoma cells L5178Y TK+/- at doses between 0.001 and 100 µl/ml, both in the presence and absence of S9 (HERA, 2005).

It can be concluded from the available evidence that CAPB at around 30 % active ingredient does not have mutagenic potential in vivo or in vitro (HERA, 2005).

Section H2.3.5 Carcinogenicity

No data relating to the carcinogenic potential of CAPB in humans or experimental species via any route of exposure were identified.

QSAR modelling could not be applied as CAPB is a zwitterion.


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Section H2.3.6 Reproductive and developmental toxicity

No data relating to the reproductive and/or developmental toxicity of CAPB in humans via any route of exposure were identified.

No experimental studies of the reproductive toxicity of CAPB were identified. As reported in section 2.3.1, no evidence of toxicity to organs, including to the ovaries and testes, was noted in a 90-day oral study at a dose of 1000 (equivalent to 300) mg/kg bw/day of a 30 % active solution (Th. Goldschmidt AG, 1991c - cited in HERA, 2005). A NOAEL of 1000 mg/kg bw/day can be derived from this study for reproductive toxicity.

In a study carried out to OECD Guideline 414 (Prenatal Development Toxicity Study), the developmental toxicity of CAPB (28.9 % active solution) was assessed in female CD rats administered doses of 0, 330, 990 and 3300 mg/kg bw/day (equivalent to 0, 95, 286 and 950 mg/kg bw/day respectively) via drinking water, on gestation days 5 – 19 (CESIO, 2004 cited in HERA, 2005). Dose-related maternal toxicity was evident as reduced body weights and stomach ulcers at doses of 990 and 3300 (equivalent to 286 and 950) mg/kg bw/day. At the highest dose, embryotoxic effects including an increased number of resorptions, decreased number of viable fetuses and decreased fetal body weight were noted. The authors derived NOAELs of 330 and 990 (equivalent to 95 and 286) mg/kg bw/day for maternal and developmental toxicity respectively (CESIO, 2004 – cited in HERA, 2005).

Section H2.3.7 Specific considerations

No other relevant studies were identified in humans or animals via any route of exposure.

Section H2.3.8 Summary of human relevant health effects

• Using lauramidopropyl betaine (LB) as a model for CAPB, absorption from the GI tract is considered to be poor.

• The acute oral toxicity of CAPB (as a 30 – 35.5 % active solution) in rats is low.

• CAPB is classified under CLP as a skin irritant (although weak) in humans. Impurities such as amidoamine may contribute to this.

• CAPB is classified under CLP as an eye irritant, with severity dependent on the percentage of active ingredient.

• CAPB is classified under CLP as a skin sensitiser (although weak) in humans. Impurities such as amidoamine may contribute to this.

• Mutagenicity and genotoxicity studies indicate that CAPB does not have mutagenic potential in vivo.

• There are no carcinogenicity data for CAPB. QSAR models predict CAPB to be non-carcinogenic.

• Subchronic oral studies indicate low toxicity of CAPB, with reversible effects following cessation of exposure.

• Reproductive toxicity studies do not show CAPB to adversely affect reproduction.

• Developmental toxicity was apparent in one OECD compliant study at maternally toxic doses of CAPB. Conversely, another study carried out to OECD Guidelines did not show fetotoxicity, even at maternally toxic doses.


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Section H3.0 Identification of relevant point(s) of


Section H3.1 Critical endpoint for human health risk assessment

purposes

The POD for use in the risk assessment (see Section 4) should, where possible, be derived from a repeated dose toxicity study, preferably via the oral route of exposure. There are no human studies relating to toxicological effects following exposure to CAPB through any exposure route. However, a number of oral repeated dose experimental studies have been carried out for CAPB, for systemic and specific organ toxicity. These are summarised in Table H3.1.


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Table H3.1 Oral route toxicity studies for CAPB



OECD 407 Repeated dose / 28 days (5 days per week) – Recovery group included for 28 days post cessation of exposure

Rat / Sprague-Dawley 250 – 1000 via oral gavage (30% active)

At the highest dose – acanthosis and oedema of the forestomach mucosa and multiple ulcerations and hyperplasia in the forestomach, with more severe findings in females. Reversible effect. No further macroscopic or microscopic effects noted in other organs, no changes seen to clinical chemistry or haematological parameters. NOAELs of 500 and 1000 mg/kg bw/day derived based on forestomach and systemic effects respectively.

Henkel KGaA, 1991 (cited in HERA, 2005)

OECD 408 Repeated dose / 90 days (5 days per week)

Rat / Sprague-Dawley 250 – 1000 via oral gavage (30% active)

At the highest dose – ulceration of the stomach at the fundus and cardia regions in males and females. At doses ≥ 500 mg/kg bw/day - forestomach gastritis with squamous hyperplasia, submucosal oedema and inflammatory cell infiltration in males and females. No further macroscopic or microscopic effects in other organs including the ovaries and testes. No changes seen to clinical chemistry or haematological parameters. NOAELs of 250 and 1000 mg/kg bw/day derived based on forestomach and systemic effects respectively. NOAEL of 1000 mg/kg bw/day derived based on reproductive toxicity.

Th. Goldschmidt AG, 1991c (cited in HERA, 2005)

OECD 414 Prenatal Development Toxicity Study / GD 5– 19.

Rat / CD 95 – 950 mg/kg bw/day via drinking water

At a dose of ≥ 286 mg/kg bw/day - maternal toxicity was evident as reduced body weights and stomach ulcers.

CESIO, 2004 (cited in HERA, 2005)


126



At a dose of 950 mg/kg bw/day - embryotoxicity apparent as an increased number of resorptions, decreased number of viable fetuses and decreased fetal body weight. NOAEL of 95 mg/kg bw/day derived for maternal toxicity. NOAEL of 286 mg/kg bw/day derived for developmental toxicity.

Shaded rows represent studies considered suitable for use in the risk assessment


127

The data indicate that the most sensitive effects following oral exposure to CAPB are forestomach effects (discussed further in section 3.3). There are no carcinogenicity studies available for CAPB. There is no evidence from currently available studies that CAPB exposure is linked to reproductive or developmental toxicity.

Section H3.2 Current health-based guidance values


Section H3.3 Identification of POD to be used for the risk

assessment

ECHA report a derived no effect level (DNEL) of 500 mg/kg bw/day. This was derived from a 28-day oral repeated dose toxicity study in rats and was based on the observed forestomach effects (Henkel KGaA, 1991 - cited in HERA, 2005).

Both the 28-day and 90-day oral repeated dose studies (section 2.3.1.) illustrate the irritant properties of CAPB on the rat forestomach, with NOAELs of 500 and 250 mg/kg bw/day respectively reported for this endpoint. However, as these effects are likely to be due to direct irritation by CAPB, possibly enhanced by gavage administration, and were found to be fully reversible, they are not considered to reflect systemic toxicity. In addition, the applicability to humans of forestomach effects in rats is uncertain.

For the reasons outlined above, the NOAEL of 1000 mg/kg bw/day derived for systemic toxicity in these studies is considered to be more representative for human health. It is noted that the CAPB test material from which the NOAEL was derived contained 30 % active substance. To reflect a 100 % solution, a NOAEL of 300 mg/kg bw/day will be used for the drinking water risk assessment (Section H4.0).

Section H4.0 Drinking water risk assessment


Few adverse effects were seen following oral exposure to CAPB.


Based on the data obtained, the NOAEL is considered to be 300 mg/kg bw/day. This is used as the POD.




AF Justification



Use of subchronic data 3 NOAEL based on a 28 and 90 day study


Total AF 600


128


Based on a POD of 300 mg/kg bw/day and an AF of 600, the proposed TDI is 500 µg/kg bw/day.




• 1000 µg/kg bw/day for a 10 kg child drinking 1 L water per day

• 667 µg/kg bw/day for a 5 kg infant drinking 0.75 L water per day

Exposure assessment

No measured drinking water concentrations were available for HEDP. The maximum concentration of CAPB measured in drinking water, as reported by WRc (2014), was 145 µg/L. This is significantly below the DWELs calculated above, indicating no risk to public health.


• 4.83 µg /kg bw/day for a 60kg adult drinking 2 L water per day

• 14.5 µg /kg bw/day for a 10kg child drinking 1 L water per day

• 21.75 µg /kg bw/day for a 5kg infant drinking 0.75 L water per day


The maximum intake of CAPB via drinking water by adults, children and infants is less than the TDI (HQ<1) and the maximum modelled concentration of HEDP in drinking water is less than the DWEL. Therefore, no adverse public health effects are anticipated following exposure to CAPB via drinking water.

Risk communication

The MOEs for CAPB, based on the NOAEL of 300 mg/kg bw/day and maximum intakes of 4.83, 14.5 and 21.75 µg /kg bw/day, are 62069, 20690 and 3908 for adults, children and infants, respectively. As the MOEs are >100, they indicate that exposures are not of concern in terms of risk to public health.

References

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Angelini G., Foti C., Rigano L., Vena GA. (1995) 3-Dimethylaminopropyl: a key substance in contact allergy to cocamidopropyl betaine. Contact Dermatitis, 32 (2):96-99. Cited in HERA, 2005.

Arimura M., Yokozeki H., Katayama I., Nakamura T., Masuda M., Nishioka K. (1998) Experimental study for the development of an in vitro test for contact allergens. Int Arch Allergy Immunol, 115 (3):228-234. Cited in HERA, 2005.


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Bonneau JC. (1990) Allergie a la Tegobetaine: a propos d´un cas. Allergie et Immunologie, 22(5), 195. Cited in HERA, 2005.

Burnett CL, Bergfeld WF, Belsito DV, et al. Final report of the Cosmetic Ingredient Review Expert Panel on the safety assessment of cocamidopropyl betaine (CAPB). Int J Toxicol. 2012;31(4 Suppl):77S-111S. doi:10.1177/1091581812447202

Cameli N., Tosti G., Venturo., N., Tosti A. (1991) Eyelid dermatitis due to cocamidopropyl betaine in a hard contact lens solution. Contact Dermatitis, 25 (4):261-262. Cited in HERA, 2005.

CESIO (2004) Prenatal development toxicity study in rats with cocamidopropyl betaine by oral administration - according to OECD guideline 414 - DRAFT. Essen, LPT Study No. 17155/03, 1-50. Cited in HERA, 2005.

ECHA Reach Registration Dossier 2020. Cocamidopropyl betaine. https://echa.europa.eu/registration-dossier/-/registered-dossier/25362/2/1 [accessed August 2020].

Foti C., Bonamonte D., Mascolo G., Corcelli A., Lobasso S., Rigano L., Angelini G. (2003) The role of 3-dimethylaminopropylamine and amidoamine in contact allergy to cocamidopropylbetaine. Contact Dermatitis, 48, 194-198. Cited by HERA 2005.

Fowler JF., Fowler LM., Hunter JE (1997). Allergy to cocamidopropyl betaine may be due to amidoamine: a patch test and product use test study. Contact Dermatitis 37 (6):276-281. Cited by HERA 2005.

Goldschmidt France S.A. (1987) TEGO Betain L7, batch 9775. Micronucleus test (Schmid method). Report No. 703201 3 March 1987, 1-18. Cited in HERA, 2005.

Henkel KGaA (1988), Banduhn N. Dehyton K - Prüfung auf Mutagenität im Ames-Test (Abschlußbericht). 880078; Feb. 1988, 1-33. Cited in HERA, 2005.

Henkel KGaA (1991) Dehyton K; 28-Tage-Test mit wiederholter oraler Verabreichung an Ratten. TED 910119; Juli 1991, 1-139. Cited in HERA, 2005.

HERA (2005). Human & Environmental Risk Assessment on ingredients of European household cleaning products; Phosphonates (CAS 6419-19-8; 2809-21-4; 15827-60-8). [Accessed July 2020].

Hunter JE., Fowler JF. (1998) Safety to human skin of cocamidopropyl betaine: A mild surfactant for personal-care products. J Surf Det 1(2):235-239. Cited in HERA, 2005.

Jackson EM (2001) The case for/against CAPB in shampoos. Cosmet Derm 14(4):60-62. Cited in HERA, 2005.

KAO Corporation (1996) Betadet HR: reverse mutation assay "Ames Test" using Salmonella Typhimurium. Haga, Tochigi, 140/473, 1-20. Cited in HERA, 2005.

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Korting HC., Parsch E-M., Enders F., Przybilla B. (1992) Allergic contact dermatitis to cocamidopropyl betaine in shampoo. J Am Acad Dermatol, 27 (6 Part 1):1013-1015. Cited in HERA, 2005.

McFadden JP., Ross JS., White IR., Basketter DA. (2001) Clinical allergy to cocamidopropyl betaine: reactivity to cocamidopropylamine and lack of reactivity to 3-dimethylaminopropylamine. Cont Derm Newsletter, 5:72-74. Cited in HERA, 2005.

Mowad CM. (2001) American Journal of Contact Dermatitis, 12 (4), 223-224. Cited in HERA, 2005.

Pigatto PD, Bigardi AS & Cusano F (1995) Contact dermatitis to cocamidopropyl betaine is caused by residual amines: relevance, clinical characteristics, and review of the literature. Am J Contact Dermat, 6:13-16. Cited in HERA, 2005.

Ross JS., White IR. (1991) Eyelid dermatitis due to cocamidopropyl betaine in an eye make-up remover. Contact Dermatitis, 25, 64. Cited by HERA, 2005.

Su L-H., Sun C-C. (1998) Positive patch test to cocamidopropyl betaine in a hairdresser. Contact Dermatitis, 38(3):168-169. Cited in HERA, 2005.

Taniguchi S, Katoh J, Hisa T, Tabata M, Hamada T (1992) Shampoo dermatitis due to cocamidopropyl betaine. Contact Dermatitis, 26, 139. Cited in HERA, 2005.

Th. Goldschmidt AG (1991c) Tego-Betain. 90 day oral (gavage) subchronic toxicity study in the rat. Essen, Th. Goldschmidt AG, 954-348-155, 26.04.1991, 1-375. Cited in HERA, 2005.

Unilever Research (1992) The fate of tegobetaine (TB) in the rat Study AM890809. Authors: Howes D, Moule RC, Fordham SJ, Curnock C, 1-53. Cited in HERA, 2005.

Van Haute N, Dooms-Goossens A (1983) Shampoo dermatitis due to cocobetaine and sodium lauryl ether sulphate. Contact Dermatitis 9, 169. Cited in HERA, 2005.

Vilaplana J, Mascaró JM, Trullás C, Coll J, Romaguera C, Zemba C & Pelejero C (1992) Human irritant response to different qualities and concentrations of cocoamidopropylbetaines: a possible model of paradoxical irritant response. Contact Dermatitis 26(5):289-294. Cited in HERA, 2005.

WRc (2014). Risk to drinking water from Personal Care Products and Domestic Cleaning Products. [Accessed July 2020].


131

4.0 Risk assessment outcomes and

conclusions

A summary of the findings from the risk assessment for each chemical of interest for adults is shown in Table 4.1 and for children and infants in Table 4.2.

Table 4.1 Risk assessment outcomes for adults

Chemical name Concentration in drinking

water (µg/L)

POD (mg/kg

bw/day)

TDI (µg/kg

bw/day)

Maximum intake (µg/kg

bw/day)

MOE HQ

1,2,3-benzotriazole 0.79 305 305 0.03 >100 0.0003

1H-benzotriazole, 4(or 5)-methyl 0.07 150 250

0.0023 >100 0.0000

(1-hydroxyethylidene) diphosphonic acid

(HEDP)

20.5 1620 1800 0.68 >100 0.0004


3.47

83

277

0.12 >100 0.0004

Amino tris(methylene phosphoric acid)

(ATMP)

20.4 1000

3333

0.68 >100 0.0002

2-(2-butoxyethoxy)

ethanol (DEGBE)

172 81

135

1.91 >100 0.0607


37

85

94

1.23 >100 0.0131

Cocamidopropyl betaine

145 300 500

4.83 >100 0.0512


132

Table 4.2 Risk assessment outcomes for children and infants


water (µg/L)

POD (mg/kg

bw/day)

TDI (µg/kg

bw/day)

Maximum intake (µg/kg bw/day)

MOE (µg/kg bw/day)

HQ

Child Infant Child Infant Child Infant

1,2,3-benzotriazole 0.79 200 305 0.079 0.1185 >100 >100 0.0008 0.0013

1H-benzotriazole, 4(or 5)-methyl 0.07 150 250

0

.007 0.0105

>100 >100

0.0001 0.0001


20.5

1620

1800

2.05 3.1

>100 >100 0.0011

0.0017


3.47

83

277

0.35 0.5

>100 >100 0.0013

0.0019

Amino tris(methylene phosphoric acid)

(ATMP)

20.4 1000

3333

2.04 3.1

>100 >100 0.0006

0.0009

2-(2-butoxyethoxy)

ethanol (DEGBE)

172 81

135

5.74 25.8

>100 >100 0.1821

0.2732


37

85

94

3.70 5.6

>100 >100 0.0392

0.0588


133


water (µg/L)

POD (mg/kg

bw/day)

TDI (µg/kg

bw/day)

Maximum intake (µg/kg bw/day)

MOE

(µg/kg bw/day) HQ

Child Infant Child Infant Child Infant

Cocamidopropyl betaine

145 300 500

14.5 21.8

>100 >100 0.1535

0.2303


134

• Hazard identification and characterisation was possible for all chemicals; however the availability of chronic toxicity studies was limited. This was reflected in the larger AF’s used to derive the HBGVs. All TDIs, with the exception of that for DEGBE, were derived from NOAELs and in several cases (HEDP, ATMP, 1,2,3-benzotriazole) this was the maximum dose tested in the study. This means that the derived ADI for these chemicals may be conservative in nature, with the ‘true’ NOAEL possibly being higher than the doses tested in the studies. A BMDL was available for DEGBE, which is the preferred POD for use in risk assessment as this approach takes into account all the available data points. This was modelled by the US EPA and based on data from a sub-chronic (90-day) study.

• Where available, the exposure assessment utilised measured data reported by WRc (2014). However, when measured data were unavailable the maximum predicted levels of each chemical in drinking water were used, as a worst-case scenario, for risk assessment purposes. The exposure assessment phase is generally considered to be the weakest part of the risk assessment process. For the three chemicals (1,2,3-benzotriazole, 1H-Benzotriazole, 4(or 5)methyl-, and LAS) where measured data were available, the measured levels were in line with the minimum predicted concentration. Use of the maximum predicted drinking water levels in the risk assessment of the remaining chemicals is therefore conservative in nature.

• The risk characterisation phase calculated the potential intake of each chemical from drinking water by receptors in three age groups. This was to take into account the effect of body weight on intake levels, with infants being seen as potentially the most sensitive age group for risk assessment purposes. Intake levels were compared against the derived TDIs which are indicative of the concentration of the substance that may be ingested over a lifetime without appreciable risk to health. For all chemicals, the estimated intake was below the TDI, and the HQ was <1.

• In addition, all DWELS were higher than measured concentrations, where available, or maximum intake levels, both previously reported (WRc, 2014).

• For each chemical and age group the MOE was also calculated to illustrate the degree of ‘safety’ between estimated intake levels and the levels associated with adverse effects. For noncarcinogenic compounds (as for those in this study) the US EPA stated that an acceptable MOE based on a NOAEL is 100 (US Environmental Protection Agency, 2012). The lowest MOE calculated in this study was 3908 for infants exposed to CAPB.

• Taken together, the evidence presented in this risk assessment indicates that the levels of PCP and DCP chemicals of interest that are potentially present in drinking water, are not anticipated to pose an appreciable risk to public health. The MOEs are sufficiently large for all chemicals to allow for additional intakes through use of the products.


135

Annex A – Literature search strategy

Table A1: Search terms and exclusion criteria

Chemical name CAS No. Search String Exclusion criteria

1,2,3-benzotriazole 95-14-7 ((“95-14-7”[Title/Abstract] OR “202-394-1”[EC/RN Number] OR “1,2,3-benzotriazole”[Title/Abstract] OR "1H-benzotriazole"[Title/Abstract]) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND (English[Language]).

Duplicates Not toxicity Aircraft Aquatic Fish Microalgae Plant Daphnia Anticancer Cytotoxic Chordata, Ascidiae Wastewater

1H-benzotriazole, 4(or 5)-methyl

29385-43-1 ((“29385-43-1”[Title/Abstract] OR “249-596-6”[EC/RN Number] OR “1H-benzotriazole, 4(or 5)-methyl-”[Title/Abstract] OR "Methyl-1H-benzotriazole"[Title/Abstract] OR “1-methyl-1H-1,2,3-benzotriazole"[Title/Abstract] OR “1H-Benzotriazole, 4(5)-methyl-”[Title/Abstract] OR “4(or5)-methyl-1H-benzotriazole”[Title/Abstract])) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND (English[Language].

Duplicates Not toxicity Aircraft Aquatic Fish Microalgae Plant Daphnia Anticancer Cytotoxic Chordata, Ascidiae Wastewater

(1-hydroxyethylidene) diphosphonic acid (HEDP))

2809-21-4

(("2809-21-4"[Title/Abstract] OR "220-552-8"[EC/RN Number] OR "HEDP"[Title/Abstract] OR "Etidronic Acid"[Title/Abstract] OR "(1-hydroxyethylidene diphosphonic acid "[Title/Abstract] OR "1-hydroxy-1-phosphonoethyl phosphonic acid"[Title/Abstract] OR "(1-Hydroxyethane-1,1-diyl)bis(phosphonic acid)"[Title/Abstract])) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND English[Language].

No abstract Duplicates Not in English Therapy Treatment Bone


15827-60-8

((“15827-60-8”[Title/Abstract] OR “239-931-4”[EC/RN Number] OR “DTPMP”[Title/Abstract] OR "Diethylenetriamine penta(methylene phosphonic acid)"[Title/Abstract] OR “Phosphonic acid, P,P',P'',P'''-[[(phosphonomethyl)imino]bis[2,1-ethanediylnitrilobis(methylene)]]tetrakis-“[Title/Abstract]) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR

No abstract Duplicates Not in English Bone


136

Chemical name CAS No. Search String Exclusion criteria

teratogen*[Title/Abstract])) AND (English[Language]).

Amino tris(methylene phosphoric acid) (ATMP)

6419-19-8 ((“6419-19-8”[Title/Abstract] OR “229-146-5”[EC/RN Number] OR “ATMP”[Title/Abstract] OR "Aminotris(methylene phosphonic acid)"[Title/Abstract] OR “Phosphonic acid, P,P',P''-[nitrilotris(methylene)]tris-"[Title/Abstract] OR “Aminotri(methylene phosphonic acid)”[Title/Abstract] OR “(1-Hydroxyethane-1,1-diyl)bis(phosphonic acid)”[Title/Abstract]) OR “nitrilotrimethylenetris(phosphonic acid)”[Title/Abstract]) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND (English[Language]).

No abstract Duplicates Not in English Therapy Advanced Therapy


112-34-5 ((“112-34-5”[Title/Abstract] OR “203-961-6”[EC/RN Number] OR “2-(2-butoxyethoxy)ethanol”[Title/Abstract] OR “DEGBE”[Title/Abstract] OR "2-(2-butoxyethoxy)ethan-1-ol"[Title/Abstract]) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND (English[Title/Abstract]):

Not toxicity Aquatic


68411-30-3 ((“68411-30-3”[Title/Abstract] OR “270-115-0”[EC/RN Number] OR “Linear alkylbenzene sulphonate”[Title/Abstract] OR "Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts"[Title/Abstract] OR “Benzenesulfonic acid"[Title/Abstract] OR “Sodium LAS”[Title/Abstract] OR “Na LAS”[Title/Abstract]) OR “Sodium alkylbenzene sulfonate”[Title/Abstract] OR “Linear alkylbenzenesulfonate, sodium salt”[Title/Abstract]) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND (English[Language].

Not toxicity Aquatic Microalga

Cocamidopropyl betaine (cosmetic grade) Cocamidopropyl betaine (technical grade)

61789-40-0; 83138-08-3; 86438-79-1 4292-10-8

((“61789-40-0”[Title/Abstract] OR “263-058-8”[EC/RN Number] OR “Cocamidopropyl betaine”[Title/Abstract]) AND ((*toxic*[Title/Abstract] OR mutagen*[Title/Abstract] OR carcinogen*[Title/Abstract] OR teratogen*[Title/Abstract])) AND (English[Language].

Not toxicity Aquatic Microalga


137


138

Annex B – Secondary screening

Table B1 below provides details of the primary literature identified for secondary screening (see section 2.0). Note that for 1H-benzotriazole, 4(or 5)-methyl and ATMP, all titles were excluded during initial screening. Table B1: Primary literature identified for secondary screening

Chemical name Publication details

1,2,3-benzotriazole Ren, Y., Zhang, L., Zhou, C-H., Geng, R-X. (2014) Recent development of benzotriazole-based medicinal drugs. Medicinal chemistry, 4, 640.

Sills, R., Hailey, J., Neal, J., Boorman, G., Haseman, J., Melnick, R. (1999) Examination of low-incidence brain tumor responses in F344 rats following chemical exposures in National Toxicology Program carcinogenicity studies. Toxicologic pathology, 27, 589-599.


Ballal, N., Das, B., Rao, S., Zehnder M, Mohn D. (2019). Chemical, cytotoxic and genotoxic analysis of etidronate in sodium hypochlorite solution. International endodontic journal, 52, 1228-1234.

Zhou, Y., Beyene, D., Zhang, R., Kassa, A., Ashayeri, E., Sridhar, R. (2008) Cytotoxicity of etidronic acid to human breast cancer cells. Ethnicity & disease 18(2 Suppl 2), S2-87-92.


US EPA. (1984) Health effects assessment for glycol ethers. Available at: https://nepis.epa.gov/Exe/ZyPDF.cgi/2000DAN7.PDF?Dockey=2000DAN7.PDF

Dugard, P., Walker, M., Mawdsley, S., Scott, R. (1984) Absorption of some glycol

ethers through human skin in vitro Environmental health perspectives, 57, 193-197.

Goen, T., Korinth, G., Drexler, H. (2002) Butoxyethoxyacetic acid, a biomarker of exposure to water-based cleaning agents. Toxicology letters, 134, 295-300.

Kerhsaw, Y. (2000) Risk assessment and risk reduction strategies PPCJ, Polymers Paint Colour Journal, 190.

Laitinen, J., Pulkkinen, J. (2005) Biomonitoring of 2-(2-alkoxyethoxy)ethanols by analysing urinary 2-(2-alkoxyethoxy)acetic acids. Toxicology letters, 156, 117-126.

Lorber, M. (1972) Hemotoxicity of synergized pyrethrin insecticides and related chemicals in intact and totally and subtotally splenectomized dogs. Acta Hepato-Gastroenterologica, 19, 66-78.

Schuler, R., Hardin, B., Niemeier, R., Booth, G., Hazelden, K., Piccirillo, V., Smith, K. (1984) Results of testing fifteen glycol ethers in a short-term in vivo reproductive toxicity assay. Environmental health perspectives, 57, 141-146.


139



Agarwal, C., Mathur, A., Gupta, B., Singh, A., Shanker, R (1990) Synthetic detergents induced-biochemical and histological changes in skin of guinea pigs. Zeitschrift fur die Gesamte Hygiene und ihre Grenzgebiete, 36, 316-318. Cresswell, D., Baldock, G., Chasseaud, L., Hawkins, D. (1978) Toxicology studies of linear alkylbenzene sulphonate (LAS) in rhesus monkeys. II. The disposition of [14C] LAS after oral or subcutaneous administration. Toxicology 11(1): 5-17. Glowienke, S., Frieauff, W., Allmendinger, T., Martus, H., Suter, W., Mueller, L. (2005) Structure-activity considerations and in vitro approaches to assess the genotoxicity of 19 methane-, benzene-and toluenesulfonic acid esters. Mutation research, 581, 23-34. Heywood, R., James, R., Sortwell, S. (1978) Toxicology studies of linear alkylbenzene sulphonate (LAS) in rhesus monkeys. I. Simultaneous oral and subcutaneous administration for 28 days. Toxicology, 11, 245-250. Mathur, A. (2005) Effects of dermal application of chromium and linear alkylbenzene sulphonate alone and in combination in guinea pigs. Toxicology International, 12, 9-12. Mathur, A., Raizada, R., Srivastava, M., Singh, A. (2005). Effect of dermal exposure to paraphenylenediamine and linear alkylbenzene sulphonate in guinea pigs. Biomedical and environmental sciences, 18, 238-240. Palmer, A., Readshaw, M., Neuff, A. (1975) Assessment of the teratogenic potential of surfactants. Part III--dermal application of LAS and soap. Toxicology 4, 171-181. Tusing, T., Paynter, O., Opdyke, D. (1960) The chronic toxicity of sodium alkylbenzene-sulfonate by food and water administration to rats. Toxicology and applied pharmacology, 2, 464-473.

Cocamidopropyl betaine Bailey, K., Tilton, F., Jansik, D., Ergas, S., Marshall, M., Miracle, A., Wellman, D. (2012) Growth inhibition and stimulation of Shewanella oneidensis MR-1 by surfactants and calcium polysulfide. Ecotoxicology and environmental safety, 80, 195-202.

Benassi, L., Bertazzoni, G., Magnoni, C., Rinaldi, M., Fontanesi, C., Seidenari, S. (2003) Decrease in toxic potential of mixed tensides maintained below the critical micelle concentration: an in vitro study. Skin pharmacology and applied skin physiology, 16 156-164.

Benassi, L., Bertazzoni, G., Seidenari, S. (1999) In vitro testing of tensides employing monolayer cultures: a comparison with results of patch tests on human volunteers. Contact dermatitis, 40, 38-44.

Burnett, C., Bergfeld, W., Belsito, D., Hill, R., Klaassen, C., Liebler, D., Marks, J. Jr., Shank, R., Slaga, T., Snyder, P., Andersen, F. (2012) Final report of the Cosmetic Ingredient Review Expert Panel on the safety assessment of cocamidopropyl betaine (CAPB). International journal of toxicology, 31, 77S-111S.


140


Cvikl, B., Lussi, A., Gruber, R. (2015) The in vitro impact of toothpaste extracts on cell viability. European journal of oral sciences, 123, 179-185.

Leidreiter, H., Gruning, B., Kaseborn, D. (1997) Amphoteric surfactants: processing, product composition and properties. International Journal of Cosmetic Science, 19, 239-253.

Suuronen, K., Pesonen, M., Aalto-Korte, K. (2012) Occupational contact allergy to cocamidopropyl betaine and its impurities. Contact dermatitis, 66, 286-292.

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