Food for Thoughts and Plastic for Dinner: the Latest ......©2015 Waters Corporation 1 Food for Thoughts and Plastic for Dinner: the Latest Analytical Landmarks in the Field of Contact

©2015 Waters Corporation 1

Food for Thoughts and Plastic for Dinner: the

Latest Analytical Landmarks in the Field of

Contact Material Contamination


Overview

Extractables & Leachables Analysis

– Definition – Workflows – Directives – Simulants

– Incidents

Example #1: Analysis of Primary Aromatic Amines (PAA)

using the ACQUITY H-Class - Xevo TQ-S micro

Example #2: Analysis of N-nitrosamines in Saliva

using the ACQUITY UPLC I-Class - Xevo TQ-S micro

Example #3: Analysis of Polymer Additives in Simulated Migration Solvents

using the ACQUITY UPLC – SQD2

Example #4: Analysis of Polymer Additives in Polymer matrix using the ACQUITY UPLC – QDa

Example #5: Analysis of Phthalates in Beverages using the ACQUITY H-Class - Xevo TQD

Example #6: Analysis of Bisphenols A, B & E in Baby Food and Infant Formula

using the ACQUITY UPLC – Xevo TQD


Definitions

Leachables are chemical entities, both organic and inorganic, that migrate from

components of a container closure system or device into a drug/food product over the

course of its shelf-life. In other words, leachables are chemical species that make their way

into the product under normal application conditions.

Extractables are chemical entities, both organic and inorganic, that will migrate from

packaging or container materials into the contents when exposed to certain solvents under

exaggerated temperature and time conditions. They are used to identify and quantify

potential leachables.

Typical extractables include monomers and oligomers from incomplete polymerization

reactions, plasticizers, stabilizers, fillers, coloring agents, antioxidants, and antistatic

agents, as well as their degradants. Additionally, residues from detergents and mold

release agents that can be present on the resin after the molding process.


Food/Pharmaceutical manufacturing equipment

– Belts, gaskets, lubricants, etc.

Food/Pharmaceutical packaging

– Paper, plastic, carton board, glass, etc.

Dining wares

– Cutlery, bowl, plate, etc.

Toys

Balloons

Cosmetics

Contact Materials/ Substances


Workflows

Identification of extractables from packaging

material

– Sample extraction

– Targeted screening for known or expected

components and their degradants (IAS)

using HR-MS

– Unknown screening of unidentified

components (NIAS) using HR-MS

Targeted analysis of extractables from

packaging material

– Migration experiments using simulated

migration solvents or beads (to mimic a

worst case scenario)

– Targeted analysis using TQ-MS

Compound name: Aniline

Correlation coefficient: r = 0.999930, r^2 = 0.999859

Calibration curve: 11380.5 * x + -898.041

Response type: External Std, Area

Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

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http://www.google.be/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0CAUQjRxqFQoTCNPMo8yMisgCFUhuFAodgOcAhg&url=http://publications.jrc.ec.europa.eu/repository/bitstream/JRC51601/guidelines test conditions_final_ed2009.pdf&psig=AFQjCNFXVXOfk9u9TrjxyNujSmil4fBefQ&ust=1442992613412234

http://www.google.be/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0CAUQjRxqFQoTCNPMo8yMisgCFUhuFAodgOcAhg&url=http://publications.jrc.ec.europa.eu/repository/bitstream/JRC51601/guidelines test conditions_final_ed2009.pdf&psig=AFQjCNFXVXOfk9u9TrjxyNujSmil4fBefQ&ust=1442992613412234


NIAS

Non-intentionally added substances (NIAS) are chemical compounds that are present in a

material but have not been added for a technical reason during the production process.

Their presence in food/pharma contact materials is generally not known by the consumer

and often is a challenge for the producer.

NIAS originate from break-down products of food/pharma contact materials, impurities of

starting materials, unwanted side-products and various contaminants from recycling

processes.


Quantitative Analyses

Photo-initiators in ink

– In-situ, after migration

– 2005: Ink curing agent Isopropylthioxanthone (ITX) detected in cardboard packaged milk

Primary Aromatic Amines in textile, printed ink, kitchenware

– In-situ, after migration

– 2007: Poorly manufactured polyamide utensils leach carcinogenic amines

Polymer additives

– In-situ, after migration from packaging

N-nitrosamines

– After migration from paint, cosmetics, balloons

Bisphenol A

– Recent concern involving infant feeding bottles

2-ethylhexyl-phthalate

– 2011: Clouding agent in probiotics substituted by probable carcinogenic compound in order to cut costs

Epoxydised soy bean oil (ESBO)

– 2005: Swiss survey of jarred foods identified concentrations exceeding the TDI allowance


Directive No 10/2011

Commission Regulation (Eu) No 10/2011 of 14 January 2011 on Plastic Materials

and Articles Intended to Come into Contact with Food

This Directive concerns packaging materials which are in direct contact with foodstuffs. This

packaging must be such that it does not transfer a hazardous part of its constituents

into/onto the food thus avoiding contamination, which could lead to adverse effects on both

human health and/or the quality of the food.

This transfer, known as migration, indicates to what extent the material is inert and

therefore a safe product. A specific migration limit (SML) is applicable for each substance. It

is expressed in mg substance per kg food. It is indicated ND if the substance shall not

migrate in detectable quantities.

In many cases a SML of 0.05 mg/kg applies. For substances for which no specific migration

limit is provided, a generic specific migration limit of 60 mg/kg shall apply.


Food Simulants

Food simulants A, B and C are assigned for foods that have a hydrophilic character and are able to extract hydrophilic substances. Food simulant B shall be used for those foods which have a pH below 4.5. Food simulant C shall be used for alcoholic foods with an alcohol content of up to 20 % and those foods which contain a relevant amount of organic ingredients that render the food more lipophilic.

Food simulants D1 and D2 are assigned for foods that have a lipophilic character and are able to extract lipophilic substances. Food simulant D1 shall be used for alcoholic foods with an alcohol content of above 20 % and for oil in water emulsions. Food simulant D2 shall be used for foods which contain free fats at the surface.

Food simulant E (Tenax) is assigned for testing specific migration into dry foods.


Xevo TQ & SQ Portfolio

Xevo TQD

Xevo TQ-S

Xevo TQ-S micro

SQD2

ACQUITY QDa


Example #1: Analysis of Primary Aromatic Amines (PAA)

using the ACQUITY H-Class - Xevo TQ-S micro


Background

The term “primary aromatic amines” (PAA) denotes a group of chemical compounds whose

simplest version is Aniline.

PAA are substances that are used, for example, in the production of certain colorants, so-

called azo pigments, notably in the color range yellow - orange - red.

Whereas a large number of PAA are safe in this respect, some PAA are known human

carcinogens. On the basis of studies involving animal experiments, others are seen as

potentially carcinogenic for humans.

For kitchenware, paper napkins and baker’s bags with colorful print and other printed food

contact items, some PAA may pose a health risk, if they are transferred to food.


Guidelines - Simulant

As regards polyamide kitchenware, the specific migration limit (SML) for PAAs is set as non-

detectable except for those on the positive list of the relevant legislation. They should not

release into foods or food simulants PAAs in a detectable quantity.

In accordance with Regulation (EU) No. 10/2011 on plastic materials and articles intended

to come into contact with food, the transfer of PAA that have not been assessed specifically,

must not be detectable in the sum.

Therefore a group SML of 0.01 mg/kg has been assigned.

As of 01/01/2013 the rules under Regulation (EU) No 10/2011 will apply. These provisions

include that for primary aromatic amine migration from polyamide kitchenware only one

migration test will be carried out.

The test is conducted with simulant B: 3% (w/v) acetic acid, as it has been demonstrated

that this simulant represents the worst case for the migration of PAAs from polyamide

kitchenware.


Compounds - Samples

A mix of 24 PAA in methanol was provided at a concentration of 100 µg/mL. Structures for a

small selection of PAA is given below.

23 out of 24 compounds could be detected at the desired levels. 3-chloro-o-toluidine was not

detected. No information is available in literature suggesting sensitivity is a common issue for

this compound.

9 extracts of kitchenware in 3% acetic acid were provided for analysis

Compound Mass Structure

Aniline 93

o-Toluidine 107

2,4-Diaminotoluene

122


Additional Sample Preparation

Some of the compounds are small, basic compounds, which are ionized with low pH. As a

result of their basic properties and the acidic sample solvent, some PAA don’t focus well on

the head of the column, resulting in poor peak shape and/or loss of retention.

Using Ammonium Hydroxide added to the 3% Acetic Acid samples prior to injection, the pH

of the sample is increased and the polar and weakly basic PAA such as Aniline will be in its

neutral form.

We believe this approach is preferred over the use of ion-pair reagent, currently described

in literature.

Sample X Sample X neutralized with NH4OH


A generic gradient was applied to separate the 23 PAAs

Adding Formic Acid to the mobile phase is not advised as it leads to loss

of retention of basic compounds on a reversed phase column. However,

using post-column addition of Formic Acid, a significant increase in

sensitivity could be obtained. The Xevo TQ-S micro integrated

fluidics simplify and fully automate this post-column addition.

UPLC Method

ACQUITY UPLC HSS T3, 1.8 µm, 2.1 x 100 mm

Mobile Phase A Water

Mobile Phase B Methanol

Column Temp 45ºC

Sample Temp 10ºC

Flow Rate 0.4 ml/min

Run Time 10 min

Injection volume 20 µL

Gradient

0 min 5% B

10 min 100% B

12 min 100% B

12.01 min 5% B

15 min 5% B


Typical Chromatogram

Overview of the 23 primary aromatic amines which could be quantified

The concentration of the compounds depicted equals 6.2 ppb (20 µL injection)


Typical Calibration Curve

Calibration curve from 0.8 to 100 ppb

Example of Aniline

Compound name: Aniline





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LOQ

Signal to Noise was calculated on 0.8 ppb standard (20 µL injection)

Compound S/N ratio

LOQ (ppb)

Aniline 377.4 0.021

o-Toluidine 768.1 0.010

2,4-Diaminotoluene 52.4 0.149

o-Anisidine 89.2 0.087

4-Chloroaniline 322.5 0.024

3-Chloro-o-toluidine ND ND

2,4,5-Trimethyl aniline 692.7 0.011

2-Methoxy-5-methylaniline

1444.3 0.005

4-Chloro-2-methylaniline 3503.1 0.002

2-Amino naphthalene 1858.2 0.004

2-Methyl-5-nitroaniline 26.6 0.293

4-Aminobiphenyl 225.5 0.035

2-Aminobiphenyl 271.6 0.029

Benzidine 559.3 0.014

4-Phenyl azoaniline 1930.7 0.004


LOQ

23 out of 24 compounds could be quantified well below the requested levels, with

quantitation limits ranging from 3 ng/L to 300 ng/L, taking into account an injection

volume of 20 µL. 19 out of the 23 compounds have detection limits below 30 ng/L. 3-

chloro-o-toluidine could not be detected, and this is confirmed by literature references.

These sensitivity levels meet the requirements for this assay, i.e. individual quantification

limits of 3 ng/mL per PAA and a total LOQ of 10 ng/mL for total PAA content.

Compound S/N ratio LOQ (ppb)

4,4’-Diamino diphenylmethane

1353 0.006

4,4’-Oxydianiline 311.6 0.025

3,3’-Dimethyl benzidine 164.5 0.047

4,4’-Thiodianiline 2582.3 0.003

o-Amino azotoluene 1745.5 0.004

3,3’-Dimethyl-4,4’-diaminodiphenylmethane

1818.2 0.004

3,3’-Dimethoxy benzidine 528.1 0.015

3,3’-Dichloro benzidine 925.9 0.008

4,4’-Methylene bis (2-chloroaniline)

1522.3 0.005


Repeatability

Repeatability (n=6) was assessed at different concentration levels: 0.4 – 0.8 – 3.1 – 12.5 –

100 ppb

The assay was found to be repeatable with RSD values (n = 6) generally lower than 5%.


Conclusion

23 out of 24 compounds could be detected at the desired levels. 3-chloro-o-toluidine was not

detected.

Using:

– Ammonium Hydroxide to neutralize the acidic samples

– post-column addition of Formic Acid to increase the signal intensity for some critical cpds

a sensitive assay was developed that can reach low ng/L concentrations

The assay was found to be repeatable with %CV values in general lower than 5%.

The samples were all below detection limits except for Aniline which was detected at 0.3 – 0.5

ng/mL.


Example #2: Analysis of Nitrosamines in Saliva

using the ACQUITY UPLC I-Class - Xevo TQ-S micro


Background

N-Nitroso compounds (NOC) are amongst the most potent carcinogens.

N-nitrosamines are used in the manufacturing of cosmetics, pesticides, and in most rubber

products. N-nitrosamines occur in latex products such as balloons, and in many foods and

other consumables.

Primary, secondary and tertiary amines can all be nitrosated to generate nitrosamines. The

secondary amines in general are the most reactive compounds towards nitrosating agents,

generating nitrosamines.

Other sources of N-nitrosamines are

– Elastomers (e.g. rubbers, silicones and thermoplastic elastomers)

– N-nitrosamines may be formed in finger paints under certain acidic conditions if they

include N-nitrosatable substances


Guidelines

Elastomers are extracted with a saliva solution containing nitrite, as ingestion will be

primarily via the mouth.

Finger paints are extracted with

– Water, simulating skin absorption

– Saliva solution containing nitrite, simulating ingestion

SML are in place for individual and the sum of all N-nitrosamines


Sample Analysis

5 nitrosamines and 1 internal standard were used for setting up a quantitative assay.

7 saliva samples were provided by the customer

ESI /EScI / APcI were compared

Linearity, sensitivity and repeatability were investigated


UPLC Method

ACQUITY UPLC I-Class system (Sample manager FTN)

Mobile phases

– Solvent A: water + 2 mM CH3COONH4

– Solvent B: methanol

– Sample manager wash solvent : Solvent B

Flow rate: 0.4 mL/min

Column temperature: 55°C

Injection volume: 30 µL

Column: ACQUITY UPLC CSH C18, 2.1 x 100 mm, 1.7 µm

Analysis time: 6 min

Gradient: from 5%B to 100%B in 4 minutes, after an isocratic hold of 30s.



Chromatogram of a 10 ng/mL standard. Compound names are depicted in the top

right corner of each trace. MRM transitions have been scheduled to maximize dwell time (15 points across a peak)


Ionization Source

NDBzA and NDBA are better in ESI,

IonSabre2 APcI is better for the small compounds like NDMA, NDEA and NePhA

IonSabre2 APcI is better than the combined EScI mode for NDMA, NDPhA and NDEA.

Because NDMA and NDEA are the least sensitive compounds, IonSabre2 APcI is the

preferred ionization mode.

The IonSabre2 APcI probe can be mounted by simply screwing the probe on the source

housing. No tools are needed.


Linearity

Calibration curve (0.1 – 10 ng/mL) for NDBA

Excellent linear correlation with r2 value of 0.9997 and % residuals of maximum 7.5%

Compound name: NDBA





ng/mL-0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0

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S/N at the 0.1 ng/mL level

The S/N for a 30 µL injection of the 5 nitrosamines

S/N based on pk-to-pk noise, no extra processing

Sensitivity


Repeatability

The repeatability (n = 6) of the quantitative analysis was determined at the level of 0.5 ng/mL

and 10 ng/mL. The % RSD’s are:


Conclusions

The nitrosamine assay on the Xevo TQ-S micro is sensitive with detection limits at and below the

desired 0.1 ng/mL level.

The assay is both linear and repeatable.

IonSabre2 APcI is the best ionization source for this multi-residue application.


Example #3: Analysis of Polymer Additives in Simulated Migration

Solvents using the ACQUITY UPLC – SQD2

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0CAcQjRw&url=http://amchamfinland.com/intertek-finland/&ei=q_MaVayALJDlar2egLAF&bvm=bv.89744112,d.cWc&psig=AFQjCNFGUq57ih1SYPW1ce6NQOrzvYh0-Q&ust=1427916038650880



Background

Plastics are made of monomers and other starting substances which are chemically reacted

to a macromolecular structure, the polymer, which forms the main structural component of

the plastics.

To the polymer, additives are added to achieve defined technological effects. The polymer

as such is an inert high molecular weight structure, and usually cannot be absorbed in the

body.

Potential health risk may occur from non- or incompletely reacted monomers or other

starting substances or from low molecular weight additives which are transferred into food

via migration from the plastic food contact material.

Therefore monomers, other starting substances and additives should be risk assessed and

authorized before their use in the manufacture of plastic materials and articles.


Sample Analysis: Objectives

Demonstrate the performance characteristics to quantify polymer additives

At levels of 10 ng/mL in 3% acetic acid and 10% alcohol

At levels of 6 µg/mL in olive oil

This report focuses on the results of the analysis of 10 polymer additives in terms of sensitivity,

linearity and repeatability.

The following Polymer Additives were analyzed: Irgafos 168, Irgafos 168 phosphate, Irganox

1010, Irganox 3114, Irganox 1076, Irganox 1330, Oleamide, Erucamide, PS800, PS802


Instrument Settings

UPLC Column: ACQUITY UPLC BEH Phenyl 1.7 µm, 2.1 mm x 100 mm

UPLC Parameters

MS settings:

– Auto dwell times (15 points across peak) and pos/neg switching in APCI mode

– All compounds in APCI positive ion mode except for Irganox 1330 (APCI-)

– RT windows according to elution time

– One m/z for quantitation and one m/z (in-source fragment) for confirmation (where applicable)

Mobile Phase A 0.1% formic acid in water

Mobile Phase B Methanol

Column Temp 40ºC

Sample Temp 15ºC

Flow Rate 0.4 mL/min

Run Time 8.5 min

Injection V 2 or 20 µL

Gradient 0 min 80% B

6 min 100% B (6)

7 min 100% B (6)

8.5 min 80% B (1)

[MH]+

N2 molecules

[Fragment]+

Collision Area

10-4

mbar

~1 mbar



100 ng/mL - Compound names are denoted on the right side


Acetic Acid & Alcohol

Excellent linearity for all compounds, in both 3% HOAc and 10% alcohol. The residual error

on the calibration plot below 10% for virtually all measured points.

The repeatability of the 6 QC’s of both 10 ng/mL and 50 g/mL standard levels was very good

on all measured concentrations (RSD < 10%).

The instrument sensitivity was enough to provide a S/N of min. 10 (required for LOQ) for a

standard at 5 ng/mL. This is half the required concentration.

IonSabre2 APcI is the best ionization source for this multi-residue application.


Typical Calibration Curve

Calibration curve and residuals for Irganox 1330

Excellent linearity (r2 = 0.999) and % residuals (all below 5.3%)

Compound name: 1330

Correlation coefficient: r = 0.999547, r̂ 2 = 0.999093

Calibration curve: 332.424 * x + 299.498



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Repeatability at 10 ng/mL

Most % RSD values below 5%


Olive Oil

Polymer Additives analysis in diluted olive oil is possible at the requested level of 6 µg/mL.

The confirmation trace is a good way to increase the selectivity for some compounds in this

complex matrix (see chromatograms for PS800).

min3.450 3.500 3.550 3.600 3.650 3.700 3.750 3.800 3.850 3.900 3.950 4.000 4.050 4.100 4.150 4.200 4.250

%

18

F2:SIR of 2 channels,AP+

329.2

20150327 008

IonSabre2 6 ug/g in olie > 60 ppb in vial

1.452e+0054.05

PS800

3.94

5023.55

3.46 3.723.53 3.593.55 3.63

4.17

min

%

1

F2:SIR of 2 channels,AP+

515.35

20150327 008

IonSabre2 6 ug/g in olie > 60 ppb in vial

3.033e+005

PS800

3.94

13865.31*

4.17


Olive Oil Spiked at 6 µg/mL

Some peak detoriation occurs (for 1076 eg)


Example #4: Analysis of Polymer Additives in

Polymer matrix using the ACQUITY UPLC - QDa

LC/MS System


http://www.google.be/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0ahUKEwi-nqCFpszMAhXDLsAKHcS8Bl8QjRwIBw&url=http://www.acesurgical.com/iv-administration/solutions-for-injection-flexible-bags/0-9-sodium-chloride.html&bvm=bv.121421273,d.ZGg&psig=AFQjCNGUBT-N_K7BQ_gG9ErH3zhL1o4ulg&ust=1462859321014838


Background

Customer has several HPLC as well as TLC methods for the detection of Polymer Additives

in extracts of the polymer itself.

The aim of this analysis was to find an alternative for the TLC analyses of the UV

transparent polymer additives:

– PS800

– PS802

– Oleamide

– Erucamide

– Stearic Acid

– Weston 618

– Hostanox SE 10

Required sensitivity levels: 10-100 ppm in AcN extract

– After polymer extraction & precipitation


Polymer Additives

Most common polymer additives are analyzed in APCI mode

– Low ppb levels can be obtained (SQD2 fit for purpose for migration purposes)

– See example #4

QDa only available with ESI probe

– Low ppm levels possible with ESI?

Experimental details

– 6 minute gradient from 80% B to 100%B

– Mobile phases:

o A = 0.02% formic acid in water

o B = methanol

– 1 µL injection volume

o Sample injected without any further sample preparation


Polymer Additives

A 10 ppm mixture of following polymer additives was analyzed (see next slide):

– Irganox PS 800

– Irganox PS 802

– Oleamide

– Erucamide

– Weston 618

– Hostanox SE10

– Stearic acid


Polymer Additives

Irganox PS 800

Irganox PS 802

Oleamide

Erucamide

Weston 618

Hostanox SE10

Stearic acid

10 ppm could clearly be detected for 6 out of 7 compounds. The observed intensities allow

for lower detection limits.

1 out of 7 compounds, Hostanox SE10, is not detected. Based on the structure ([H3C-

(CH2)17-S-]2 of the compound, it is likely that this compound is not MS sensitive at all.


Example #5: Analysis of Phthalates in Beverages

using the ACQUITY H-Class - Xevo TQD




Phthalates

Phthalates cover a large group of compounds, esters of phthalic acid Known toxic effects

− Considered endocrine disruptors, related to reproduction in animal studies Migration can occur during production and storage, from packaging materials,

coatings, equipment coatings, sealants, etc.

In 2013, China banned the importation of European spirits unaccompanied by CoA for phthalates


Challenges in phthalate analysis

Background ~7e4

DBP at 100 ppb

Traditionally, analysed by gas chromatography

– Derivatisation and/ or extraction required

– Non selective m/z 149 monitored for multiple compounds

Growing interest liquid chromatography method

– Reduced sample preparation: dilute and shoot

– Improved selectivity on m/z transitions

Ubiquitous contaminants

– Significant background impacts accurate

quantification by GC and/ or LC


Isolator column: Overview

Ubiquitous contaminants

AcQuity C18 isolator column

− Part number: 186004476

Isolator separates background

contaminants from analytes of

interest

Isolator column

Sample injector

Solvent mixer


Isolator column: Contaminant separation

BBP Background BBP contaminant


Example of spirit matrices spiked with BBP at 100 µg.l-1

Summary of Results

Sample A

Brandy

Sample B

Gin

Sample C

Whiskey

Compound name: BBPCorrelation coefficient: r = 0.997755, r^2 = 0.995515Calibration curve: 839.103 * x + 1264.79Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

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BBP

Compound name: DNOPCorrelation coefficient: r = 0.998477, r^2 = 0.996957Calibration curve: 763.204 * x + 168.408Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

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DNOP

Example of whiskey matrix matched calibration from 5 to 100 µg.l-1


Conclusions

Vast and varied area of analysis with international interest increasing

Extensive legislation in place, with some commonalities and solid base for safety and

analytical approach

Challenges faced by food industry for good communication with packaging manufacturers

Simple and quick “dilute and shoot” method for the accurate quantification of migration of

materials in contact with foodstuffs

Improves consumer confidence and ensures compliance for export with high throughput


Example #6: Analysis of Bisphenols A, B & E in Baby Food

and Infant Formula using the ACQUITY UPLC – Xevo TQD


BPA Background

What is BPA?

– BPA is the starting material for polycarbonate polymerization

– It has been an important industrial chemical not only to make polycarbonate plastic but

also epoxy resins; both of which are used in a wide variety of applications.

o Polycarbonate plastics are used in baby bottles, tableware and other food containers

• (Eyeglass lenses, medical equipment, digital media (e.g., CDs and DVDs), cell

phones, consumer electronics, computers and other electrical equipment, …)

o Epoxy resins are used in can coatings, industrial floorings, adhesives, industrial

protective coatings, powder coatings, and printed circuit boards


BPA Background

What are the potential health issues?

– Can leach into food from the epoxy resin lining of cans and from

consumer products such as polycarbonate tableware, food storage

containers, water bottles, and baby bottles.

– BPA is a weak "estrogen like" endocrine disruptor which can mimic

circulatory endogenous hormones and is suspected to be linked to a

higher incidence of certain cancers, reproductive disorders, diabetes

and heart disease in humans


International Regulations

Canada - First country to take action, listed it as a toxic substance

US-FDA - Released a report in 2010 expressing some concern regarding exposure of fetuses, infants and young children to BPA. Report encourages further studies into the safety of BPA

European Union - EU directive (2011/8/EU) has banned the use of BPA in infant feeding bottles

o France Bans Bisphenol A in All Food Contact Materials: French National Assembly has been voting to ban BPA from 2014 on. Concerning baby nutrition products the ban will step into force in 2013

Japan - Between 1998-2003, canning industry voluntarily replaced its BPA-containing epoxy resin can liners with BPA-free polyethylene terephthalate (PET) in many of its products


Column: ACQUITY UPLC BEH C18 2.1 x 50 mm, 1.7 m

Column temp 40 C

Mobile phase A 0.5 % NH4OH in H2O

Mobile phase B 0.5 % NH4OH in MeOH

Elution 3 minute linear gradient from 5% (B) to 95% (B)

Flow Rate 0.5 mL/min

Injection volume

50 L

Experimental Conditions


Part 1 – Protein precipitation

Part 2 - DisQUE

Part 3 – OASIS HLB

Optimised Sample Prep Protocol

Precipitation (10g sample, 10mL CH3CN) Centrifuge and collect supernatant

Add contents from DisQuE tube 1. Shake.

Centrifuge & collect 10 mL supernatant

Add contents from DisQuE tube 2. Shake.

Centrifuge & collect supernatant.

Dilute supernatant with 70 ml H2O. Load on Oasis HLB (3cc)

Wash – 2 mL 40 % MeOH Elute – 1 mL 100 % MeOH

Dilute eluate with 1 mL H2O Inject 50 µL for LC-MS/MS


Spiked Compounds: 1 ng/mL in Samples

S/N > 3


Correlation coef f icient: r = 0.998164, r^2 = 0.996332

Response type: Internal Std ( Ref 4 ), Area * ( IS Conc. / IS Area )

Curve type: Linear, Origin: Exclude, Weighting: 1/x^2,

Conc-0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0

Res

po

nse

-0.00

0.10

0.20

0.30

0.40

0.50

0.60

0.70

0.80

0.90

1.00

1.10 Bisphenol Infant Formula (10 pg/μL spike)

BPA 102% (3.2%)

BPB 95% (5.5%)

BPE 81% (4.6%)

Linearity and Recovery

Bisphenol Baby food

(20 pg/μL spike)

BPA 110% (7.8%)

BPB 112% (6.7%)

BPE 99% (6.1%)


Thank you for your attention


Additional Information about Legislation

Documents

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