Dr. Gisela FontaineTeamleader ICP-MS, Solvias AG

01.10.2019

Inorganic Trace Element Analysisin the Pharmaceutical Industry

• Sources of pictures, diagrams, etc. are all listed at the end of the ppp and as far as possible (some textbook knowledge) with the respective presentation, current pictures from the lab have been taken by the presenter

• Per definition, it is the PRESENTER’S PERSPECTIVE on the topic, but neither necessarily the position of Solvias nor the only way to address the task.

Disclaimer

Master Students, PhD Students,…Who are You?

Gisela Fontaine

- BSc & MSc at ETH Zürich (CH), Chemistry, Focus on Analytics

- PhD Thesis Group of Prof. Dr. Detlef Günther, ETH Zürich (CH) “Fundamental studies on mass bias variability in multi collector-inductively coupled plasma mass spectrometry and the use of isotope ratios in gem authentication”

- Lab head UFAG Laboratorien AG, Sursee (CH): AAS, ICP-OES, ICP-MS, heavy metals limit test, purified water testing, Working under GMP & ISO 17025

- Team leader Solvias AG, Kaiseraugst (CH): ICP-MS (AAS, ICP-OES, Polarography), working under GMP & ISO 9001

Who am I?

Who is Solvias?

Integrated services that enhance the value chain for customers

• Swiss Contract Research Organization (CRO) & CDMO

• Founded in 1999, majority employee and management owned

• > 500 employees

• ISO9001, GMP and GLP certified

• Successfully FDA inspected

• Sales > CHF 75 Mio. (2017)> CHF 90 Mio. (2018)

Centersof Excellence in Switzerland & France, US Subsidiary

HombourgFrance

BaselSwitzerland

KaiseraugstSwitzerland (HQ)

Small Molecule Analysis

Custom Synthesis

Ligands & Catalysis

Polymorph, Salts & Crystallization

Biopharmaceutical Analysis

DNA Analysis

Extractables & Leachables

Troubleshooting

Bio Analysis & Cell Based Bioassays

Process Analytical Technology Probes

Environmental Monitoring

Elemental & Trace Analysis

Medical Device Analytics

Inhaled Drug Products

Quality Control

What We Do

SMAL

L M

OLE

CU

LES

BIO

PHAR

MAC

EUTI

CAL

S

SYNTHESISCustom Synthesis

API SynthesisProcess R&DLigand Supply

Preclinical phase I phase II phase III commercial

SOLID STATESalt/Co-Crystal Selection

Polymorph Selection

Crystallization Development

ANALYTICALCharacterization

Method DevelopmentMethod Validation

QUALITY CONTROL

STABILITY STUDIESForced Degradation

ICH Stability

Post Approval Studies

TROUBLESHOOTING

Supporting the Value-Chain of Our Customers

Who are Our Customers?

Substances Analytical Testing Services + Products Devices

Ligands &Specialty Products

Analytical Services Confarma Biopharma Analytical

Technology

• Ligands• Organocatalysts• Porphyrines• Catalysis and

synthesis services

• Physicochemistry• QC release• Troubleshooting• Solid state• Method

development

• Cell-biology, virology

• Physicochemistry• Products / Kits• Troubleshooting

• Characterization• Stability studies• Method

development• Protein analysis

• Probes• Safety projects• SoftwareProducts

Services

• Large pharma• CMO• Distributors for

ligands

• Large pharma• Small and mid-

size comps.• CMO

• Pharma• Medical Devices• Cosmetics

• Large pharma and biotech

• Medical Devices

• Selling mostly through third party channel

Customers

CRO in the General Analytical Principle

Sample Introduction Excitation Reaction Detection

• Analytical question• Investigation object• Representative sampling• Sample preparation• Analytical sample

• Data Evaluation• Result• Interpretation • Answer• Decision / Actions

Analytical Laboratory

Analytical Instrument

API (Active Pharmaceutical Ingredient) • cis-Platinum / AsIII as cytostatic / anti-cancer agent• Gadolinium as contrast agent for MRI

Included in the final product as• Part of molecular structure of API, e.g. B, Br, Se, Zn,…• Counterion: Na, K,…• Coating: iron oxide, titanium oxide,…• Excipient: TiO2, SiO2, talcum (Mg3Si4O10(OH)2),…

Use during manufacturing (not wanted in final product)• Catalysts: Pd, Rh, Sn, Pt,…• Reagents: NaBH4, tert-Butyl-Li, SnCl2, …• Feed (for bioproduction): salts of Cu, Zn, Fe,…

Inorganic «Elemental Impurities»: what we DON’T want

ExamplesMetals – Use in the Pharmaceutical Industry

PSE

Analytical Questions Trace Elements

What is this greymatter made of?

Release Trouble-ShootingScreenings

For a 100 years…Elemental Impurities

Since 1908: visual limit test withsulfide precipitation of metals withthioacetamide, using Pb as reference, mainly covering As, Mo, Ag, Cd, Sn, Sb, Pt, Pb, Bi, (and Hg) Applied mainly at max. 10 mg/kg

N. Lewen, 2004

• Pharmacopeiae: definition of requirements for drugs and drug ingredients (+reagents, containers, …) for strength, quality and purity

• Specific SOPs (Standard operating procedures): develop, define, validate & follow• Anything which has not been noted has not been done

Ensuring drug products are safe, pure and effective

«By the book»GMP – Good Manufacturing Practice

A attributable: Who, when? L legible: readable (for a long time)C contemporaneous: instantaneous, completeO original: where? A accurate: error free, complete, true,

encompassing

ALCOA-Concept for Documentation

• 4-Eye-Principle: Review

• All data needs to be traceable all the time

• Validation of computer-based systems

• Integrity / Possibility to change electronic (raw) data

• Lifecycle of electronic data: Archiving, saving, readability of data over time

Data Integrity

Enter your password here:

Described in the pharmacopeiae as separate chaptersSpectroscopic Techniques for Element Analysis

Technique Ph. Eur. USP-NF JP

AAS Ph. Eur. 2.2.23Atomic Absorption Spectrometry

USP-NF ⟨852⟩Atomic Absorption Spectroscopy

JP <2.23>Atomic AbsorptionSpectrophotometry

ICP-OES Ph. Eur. 2.2.57Inductively Coupled Plasma-Atomic Emission Spectrometry

USP-NF <730>PlasmaSpectrochemistry / USP-NS <1730> PlasmaSpectrochemistry- Theory & Practice

JP <2.63> Inductively CoupledPlasma-AtomicEmissionSpectrometry andInductivelyCoupled Plasma-Mass Spectrometry

ICP-MS Ph. Eur. 2.2.58Inductively CoupledPlasma-Mass Spectrometry

IntroductionShort description ofmethod+ Reference to furtherinformationInstrument QualificationIQ – Installation QualificationOQ – Operational QualificationPQ – Performance Qualification(with specificrequirements)Disclaimer that specificmonographs applyprior to this chapter

USP42-NF37 1S <730> Plasma SpectrochemistrySet-Up Pharmacopeial Chapters

ProcedureMaterials & reagentsto be usedStandard solutionsto be usedTypes of calibration / quantificationSample PreparationBroad range given, reference toseparate monographsAnalysisCalibration rangeSystem SuitabilityTest (SST)

Validation & Verification«demonstrate that themeasurement is suitable forits intended purpose»Specific validation criteriaAccuracyPrecision (repeatability & Intermediate precision)Different acceptance criteriadrug substance assay, drugproduct assay and impurityanalysisSpecificity AccuracyQuantitation limit (QL/LOQ)LinearityRangeRobustness

Verification«simply verifytheir suitabilityunder actualconditions ofuse»

Possibility tovalidatealternative procedure

Contact person

A Sample Arrives

+

Pharmacopeia monograph

Product-specific customerSOP - external(standard operatingprocedure)

Product-specific Solvias SOP – internal

Solvias SOP

Analysis

CEM, Anal. Chem., 1987, 59 (4), pp 302A–302A

From open digestion to single reaction chamberSample Digestion

http://www.topautoclave.com/s/cc_images/cache_26108279.jpg?t=1403072879

https://media.labcompare.com/m/1/Product/2751880-400x300.jpg

https://milestonesci.com/wp-content/uploads/2017/07/PP_UltraCLAVE_Thumb2.png

Weigh samples, blanks & reference materials

«Metal-free» sample preparation roomKeep it clean!

Add acidsPredigest in ultrasonic bath

Digest at high temperature & pressure

4 Ultra-Clave digestion units1 TurboWave

In the labICP-MS • 3 Agilent (Q-ICP-MS) for screenings + routine analysis

• 1 Agilent triple-quad instrument for improved interference control (complex matrices & interfered analytes)

Short Excursion: more than just «one» instrumentICP-MS

11 specialists• Technicians: sample preparation, analysis,

evaluation• Scientists: validations, method development, lab

planning• Reviewer

The ICP-MS Team

• 30 - 50 analytical requests per week / 1 - 36 analytes/ 1 - 60 samples ~1500 results / week

• Ca. 30 validations / year

Your Turn-

The Analytical Challenge

Your Analytical Challenge

You are an analytical scientist in the trace element team of a contract laboratorywhich works largely according to quality standard GMP (good manufacturingpractice). Imagine you have nearly unlimited equipment available.

Case 1: TroubleshootingA customer from a large pharmaceutical company comes to you because theyhave a problem with one of their production sites where they produce a preciousAPI (active pharmaceutical ingredient): They have found a hair crack in one oftheir centrifuges and black, greasy material outside of the container. Theycontact you for help.• Why is your customer concerned? What are the potential risks?• What might the dark matter coming out of the centrifuge be composed of?• What are key considerations you should focus on as a contract lab?• Which analyses would you suggest?

- OOS: Out of specification / Deviation- Risk Assessment- Stop of production loss of money- Loss of control changing conditions- Loss of product- Contamination / degradation of product

Why is your customer concerned? What are the potential risks?

• Solvent• Grease (https://www.mobilindustrial.com/ind/english/files/tt-components-and-

characteristics-of-grease.pdf, visited 30.09.2019): • base oil: 80-97%: oil with boiling point range 550 - 1050 F, consisting of hydrocarbons with 18 to

40 Cs C, O, H• Thickener: e.g. lithium, aluminum, calcium soaps; clay (AlxSiyOz(OH)x); polyurea; alone or in

combination. • Additives: “oxidation or rust inhibitors, polymers, extreme pressure (EP) additives, anti-wear

agents, lubricity or friction-reducing agents(soluble or finely dispersed particles such as molybdenum disulfide and graphite) and dyes or pigments” (only for color).

• API and degradation products• Rust

What might the dark matter coming out of the centrifuge be composed of?

• What is the customer ready to pay for?• What can you deliver (consider equipment,

time, effort, quality, cost)• Quality standard?• Customer satisfaction: Strategic

considerations• External restrictions: available sample

material

What are key considerations you should focus on as a contract lab?

Scope

Cost

Quality

Time

• Which type of equipment do you have available?• Screening analyses:

• EDX• ICP-MS screening• API: in-house

Which analyses would you suggest?

Liquid collectedin pipette

Greasymaterial collectedoutside crack

67 mg 34 mg

Rust!

Case 2: Routine AnalysisA crack in high quality, well-maintained equipment is something, which wouldusually not occur. However, in the overall production process of an API, theremight be a range of sources for “elemental impurities” which need to becontrolled.• What are “elemental impurities” according to ICH Q3D?• What might be sources of elemental impurities in a production process?• If you had to estimate to which extent they might come out and how this could be

controlled, how would you set up a risk assessment?• If you needed to analyse the Class 1, Class 2B elements plus the used catalyst Pd for

your customer in a drug product (tablet), which is prescribed with a max. daily dose of 2 g per day, which method/s would you suggest for analysis?

• What would be a suitable sample preparation? Which considerations would be different for Tablet A and B (composition below)

• If your customer wanted to implement analysis of those elements for release analysis, how would you ensure that the method is “valid” to analyse them at this level – in compliance with quality standard GMP (good manufacturing practice)?

What are “elemental impurities” according to ICH Q3D?

Class Elements Toxicity Occurrence / Source

Evaluation during Risk Assessement

1 As, Cd, Hg, Pb High Commonly used material

All pot. sources, all admin. routes

2A Co, Ni, V Route-dependent High All pot. sources, all admin. routes

2B Ag, Au, Ir, Os, Pd, Pt, Rh, Ru, Se, Tl

Route-dependent Low Only if intentionally added (in excipient/DS or DP)

3 Ba, Cr, Cu, Li, Mo, Sb, Sn

Relatively low (oral PDE > 500 µg / day)Considerable (parenteral / inhalation)

Oral -> only if intentionally addedParenteral / inhalation -> needed unless PDE > 500 µg / day

4 Al, B, Ca, Fe, K, Mg, Na, W, Zn

No established PDE, addressed by other guidelines (particular elements need to be considered in particular products or for quality consideration)

Permitted Daily Exposure (PDE)ICH Q3D, Step 4 «Elemental Impurities»

Calculation max. permitted concentrationc via daily dose drugproduct Dd

𝑐𝑐𝜇𝜇𝜇𝜇𝜇𝜇 =

𝑃𝑃𝑃𝑃𝑃𝑃( 𝜇𝜇𝜇𝜇𝑑𝑑𝑑𝑑𝑑𝑑)

𝑃𝑃𝑑𝑑( 𝜇𝜇𝑑𝑑𝑑𝑑𝑑𝑑)

Depending on• Toxicity• Route of

administration• Bioavailability

Ishikawa-Diagram, Fishbone-Diagram

What might be sources of elemental impurities in a production process?

Erosion / Abrasion frommanufacturing materials:

Steel: Fe, Cu, Zn, Mo, (W)Aluminium parts

Unsufficient removal of- catalysts,

- (inorganic) reagents- feed metals

- processing aids

Contamination fromenvironment

AirDust

Container closure

system

Contaminated(re)agentsSolventsWater

Organics

If you had to estimate to which extent they might come out and how this could be controlled, how would you set up a risk assessment?

IDENTIFY• Review API, excipient, drug product manufacturing process for known and

potential sources of elemental impurities that may find their way into the drug product

EVALUATE• the presence of a particular elemental impurity in the DP by determining the

observed or predicted level and compare it to the PDE

DOCUMENT• Summarize and document the risk assessment• If needed, identify additional control requirements in the to limit the elemental

impurity in the DP?

If you needed to analyse the Class 1, Class 2B elements plus the used catalyst Pd for your customer in a drug product (tablet), which is prescribed with a max. daily dose of 2 g per day, which method/s would yousuggest for analysis?

- LOQs required- Sensitivity of method- Stabilization of analytes in solution- Digestion of matrix (dilution)

Cover as many analytes as possible in 1 methodChallenge

Comparison mechanism of action of spectroscopicmethods for analysis of trace elements

Main Spectrometry Methods

S. Traxel, «Metallische Verunreinigungen», 06.10.2016

Suitability of Methods for ICH Q3D Option 1 Limit Solution Concentration [μg/L] LODs / LOQs per Method [μg/L]

ElementClass Oral Parenteral Inhalation (Dilution Oral, Option 1) AAS

[µg/g] [µg/g] [µg/g] 1:10 1:50 1:500 F-AAS GF-AASCV/HG-

AAS ICP-OES ICP-MSCd 1 0.5 0.2 0.2 50 10 1 206 0.0061 - 0.21 0.051

Pb 1 0.5 0.5 0.5 50 10 1 1006 0.65 - 21 0.051

As 1 1.5 1.5 0.2 150 30 3 201 0.61 0.021 201 0.051

Hg 1 3 0.3 0.1 300 60 6 2001 155 0.0011 0.21 0.021

Co 2A 5 0.5 0.3 500 100 10 61 0.041 - 0.51 0.024

V 2A 10 1 0.1 1000 200 20 401 0.41 - 51 0.024

Ni 2A 20 2 0.5 2000 400 40 41 0.041 - 0.51 0.0051

Tl 2B 0.8 0.8 0.8 80 16 1.6 101 0.21 - 21 0.011

Au 2B 10 10 0.1 1000 200 20 61 0.21 - >0.1 <12 ≤0.0023

Pd 2B 10 1 0.1 1000 200 20 1006 0.95 - >0.1 <12 ≤0.0023

Ir 2B 10 1 0.1 1000 200 20 50006 13.55 - >0.1 <12 ≤0.0023

Os 2B 10 1 0.1 1000 200 20 10006 - - >0.1 <12 ≤0.0023

Rh 2B 10 1 0.1 1000 200 20 506 0.85 - >0.1 <12 0.00043

Ru 2B 10 1 0.1 1000 200 20 10006 1.55 - >0.1 <12 0.024

Se 2B 15 8 13 1500 300 30 1001 21 0.021 5.91 0.11

Ag 2B 15 1 0.7 1500 300 30 [11] 0.011 - >0.01 <0.12 0.24

Pt 2B 10 1 0.1 1000 200 20 401 0.41 - >0.1 <12 0.00013

Li 3 55 25 2.5 5500 1100 110 0.51 0.41 - 20 0.024

Sb 3 120 9 2 12000 2400 240 301 0.21 - >0.1 <12 0.024

Ba 3 140 70 30 14000 2800 280 101 0.081 - 0.51 0.24

Mo 3 300 150 1 30000 6000 600 301 0.041 - 0.071 0.14

Cu 3 300 30 3 30000 6000 600 11 0.041 - 11 0.011

Sn 3 600 60 6 60000 12000 1200 201 0.21 0.51 301 0.14

Cr 3 1100 110 0.3 110000 22000 2200 21 0.021 - 0.51 0.00511 K.Camman, Instrumentelle Analytische Chemie, Spektrum Akademischer Verlag, Berlin, 2001 4 generally validated LOQs Solvias ASOP-05852 icap ICP-OES brochure Thermo Fisher Scientific 5 User’s Guide Agilent Analytical Methods for Graphite Tube Atomizers3 D. Kutscher et al., Spectroscopy, 2018, 33, 9, p 16–25 6 Varian Analytical Methods for Flame AAS (lowest optimum working range)

Comparison of Analytical MethodsWorking Range Acquisition Cost

Dynamic Range Measurement Speed

Sample PreparationMeasurementslow fast

S. Traxel, «Metallische Verunreinigungen», 06.10.2016

AAS: Analytical ChallengesParameter• Non-linear calibration curves due to

difference in full width at half maximumof primary radiation from HCL andabsorption profile of atomisation unit

• Degree of atomisation, e.g. due toformation of stable species (oxides / phosphates)

• Degree of ionization for easilyionizable elements, e.g. Na, K, Rb, Cs

• Unspecific absorption e.g. bymolecules or light scattering in graphitefurnace by inorganic salts

Mitigation• Try to work in linear part of calibration

curve (adjust concentration)

• Flame: Increase heat: use N2O/C2H2, furnace: increase temperature or avoidformation by chemical intervention(modifier, e.g. Pd(NO3)2, Mg(NO3)2,…)

• intentionally add easily ionizableelement to increase partial pressure offree electrons

• Background compensation withreference radiation, e.g. by continuumsource (deuterium lamp)

concentrationextin

ctio

n

AAS: Characteristics# of Elements 70Data Aquisition Sequentially (slow)Sample Types Liquid, solid (gaseous)

LOD[μg/L]

Sample Volume [mL]

Working Range [μg/L]

Flame 100 2 – 10 100 – 100’000Graphite Furnace (GF) 0.01 – 0.2 0.01 – 0.1 0.01 – 200Hydride (As, Se, Sb, Sn) 0.02 – 0.5 50 0.02 – 10Cold Vapour (Hg) 0.2 50 0.02 – 10Dynamic Range 3 orders of magnitudeAnalytical Expertise Average (chemical expertise GF-AAS increased)Cost Aquisition Low (30’000 – 70’000 €)Running Cost Low (consumables, C2N2/air (N2O))

Comparatively cheapSelectiveSensitive (GF)

Single elementSeveral introductiontypes required

ICP-OES: Analytical ChallengesParameter• Matrix interferences difference in

sensitivity between calibration andsample solutions

• Spectral interferences due tobackground emission

• Spectral interferences of a matrix component / different analyte at same wavelength

Mitigation• Addition of internal standard to

compensate for sample take-up andplasma fluctuations (best of similarchemical characteristics) / Matrix-matched calibration

• Background compensation bymonitoring spectrum around analyticalwavelength and extrapolation + blank subtraction

• Monitor more than one wavelength todetect / avoid spectral interferences

Concentration

Inte

nsity

https://www.thermofisher.com/ch/en/home/industrial/spectroscopy-elemental-isotope-

analysis/spectroscopy-elemental-isotope-analysis-learning-center/trace-elemental-analysis-tea-information/icp-oes-information/icp-oes-data-

analysis.html

ICP-OES: Characteristics# of Elements 70Data Aquisition Simultaneously or sequentially (fast)Sample Types Liquid, solid (after digestion)Limits of Detection 0.1 – 30 μg/L (in solution)Dynamic Range 4 – 5 orders of magnitudeWorking Range 0.01 – 1000 mg/L (in solution)Analytical Expertise AverageCost Aquisition Average (60’000 – 130’000 €)Running Cost High (Ar)

Multi-elementRobustGood selectivity

Sensitivity restricted

ICP-MS: Analytical ChallengesParameter• Matrix interferences difference in

sensitivity between calibration andsample solutions (e.g. degree ofionization, mainly Se, As, Au)

• Isobaric interference at same m/z by− Isotope of another element− Polyatomic species (MO+, MAr+, MX+)− Doubly charged species M++

Mitigation• Addition of internal standard to

compensate for sample take-up andplasma fluctuations (best of similarchemical characteristics) / Matrix-matched calibration

• Don’t add/avoid interferant (if added, e.g. hydrochloric acid)

• Instrument tuning• Monitor more than one isotope/mode to

detect / avoid isobaric interferences, allowmathematical corrections

• Analyse in high resolution mode• Chemical/kinetic resolution by collision /

reaction cell technology

© 1991 IUPAC and adapted from PerkinElmerSciex „Relative Abundance of the Natural Isotopes", 2006

Relative Abundance of the Natural Isotopes

Isotope % % % Isotope

% % %

Isotope

% % %

Isotope

% % %

1 H 99.985 2 H 0.015

61 Ni 1.140 62 Ni 3.634

121 Sb 57.36 122 Sn 4.63 Te 2.603

181 Ta 99.988 182 W 26.3

3 He 0.0001 63 Cu 69.17 123 Te 0.908 Sb 42.64 183 W 14.3 4 He 99.9998 64 Zn 48.6 Ni 0.926 124 Sn 5.79 Te 4.816 Xe 0.10 184 Os 0.02 W 30.67 5 65 Cu 30.83 125 Te 7.139 185 Re 37.40 6 Li 7 Li 8

7.5 92.5

66 Zn 27.9 67 Zn 4.1 68 Zn 18.8

126 Te 18.95 Xe 0.09 127 I 100 128 Te 31.69 Xe 1.91

186 Os 1.58 W 28.6 187 Os 1.6 Re 62.60 188 Os 13.3

9 Be 100 69 Ga 60.108 129 Xe 26.4 189 Os 16.1 10 B 19.9 70 Ge 21.23 Zn 0.6 130 Ba 0.106 Te 33.80 Xe 4.1 190 Os 26.4 Pt 0.01 11 B 80.1 71 Ga 39.892 131 Xe 21.2 191 Ir 37.3 12 C 98.90 72 Ge 27.66 132 Ba 0.101 Xe 26.9 192 Os 41.0 Pt 0.79 13 C 1.10 73 Ge 7.73 133 Ce 100 193 Ir 62.7 14 N 99.643 74 Ge 35.94 Se 0.89 134 Ba 2.417 Xe 10.4 194 Pt 32.9 15 N 0.366 75 As 100 135 Ba 6.592 195 Pt 33.8 16 O 17 O 18 O

99.762 0.038 0.200

76 Ge 7.44 Se 9.36 77 Se 7.63 78 Kr 0.35 Se 23.78

136 Ba 7.854 Ce 0.19 Xe 8.9 137 Ba 11.23 138 Ba 71.70 Ce 0.25 La 0.0902

196 Hg 0.15 Pt 25.3 197 Au 100 198 Hg 9.97 Pt 7.2

19 F 100 79 Br 50.69 139 La 99.9098 199 Hg 16.87 20 Ne 21 Ne 22 Ne

90.48 0.27 9.25

80 Kr 2.25 Se 49.61 81 Br 49.31 82 Kr 11.6 Se 8.73

140 Ce 88.48 141 Pr 100 142 Nd 27.13 Ce 11.08

200 Hg 23.10 201 Hg 13.18 202 Hg 29.86

23 Na 100 83 Kr 11.5 143 Nd 12.18 203 Tl 29.524 24 Mg 25 Mg 26 Mg

78.99 10.00 11.01

84 Kr 57.0 Sr 0.56 85 Rb 72.165 86 Kr 17.3 Sr 9.86

144 Nd 23.80 Sm 3.1 145 Nd 8.30 146 Nd 17.19

204 Hg 6.87 Pb 1.4 205 Tl 70.476 206 Pb 24.1

27 Al 100 87 Sr 7.00 Rb 27.835 147 Sm 15.0 207 Pb 22.1 28 Si 29 Si

92.23 4.67

88 Sr 82.58 89 Y 100

148 Nd 5.76 Sm 11.3 149 Sm 13.8

208 Pb 52.4 209 Bi 100

30 Si 3.10 90 Zr 51.45 150 Nd 5.64 Sm 7.4 210 31 P 100 91 Zr 11.22 151 Eu 47.8 211 32 S 95.02 92 Zr 17.15 Mo 14.84 152 Gd 0.20 Sm 26.7 212 33 S 0.75 93 Nb 100 153 Eu 52.2 213 34 S 4.21 94 Zr 17.38 Mo 9.25 154 Gd 2.18 Sm 22.7 214 35 Cl 75.77 95 Mo 15.92 155 Gd 14.80 215 36 S 0.02 Ar 0.337 96 Zr 2.80 Mo 16.68 Ru 5.52 156 Gd 20.47 Dy 0.06 216 37 Cl 24.23 97 Mo 9.55 157 Gd 15.65 217 38 Ar 0.063 98 Mo 24.13 Ru 1.88 158 Gd 24.84 Dy 0.10 218 39 K 40 K 41 K

93.2581 0.0117 Ca 6.7302

96.941 Ar

99.600

99 Ru 12.7 100 Mo 9.63 Ru 12.6 101 Ru 17.0

159 Tb 100 160 Gd 21.86 Dy 2.34 161 Dy 18.9

219 220 221

42 Ca 0.647 102 Pd 1.02 Ru 31.6 162 Er 0.14 Dy 25.5 222 43 Ca 0.135 103 Rh 100 163 Dy 24.9 223 44 Ca 2.086 104 Pd 11.14 Ru 18.7 164 Er 1.61 Dy 28.2 224 45 Sc 100 105 Pd 22.33 165 Ho 100 225 46 Ti 8.0 Ca 0.004 106 Pd 27.33 Cd 1.25 166 Er 33.6 226 47 Ti 7.3 107 Ag 51.839 167 Er 22.95 227 48 Ti 73.8 Ca 0.187 108 Pd 26.46 Cd 0.89 168 Er 26.8 Yb 0.13 228 49 Ti 5.5 109 Ag 48.161 169 Tm 100 229 50 Ti 5.4 V 0.250 Cr 4.345 110 Pd 11.72 Cd 12.49 170 Er 14.9 Yb 3.05 230 51 V 99.750 111 Cd 12.80 171 Yb 14.3 231 Pa 100 52 Cr 83.789 112 Sn 0.97 Cd 24.13 172 Yb 21.9 232 Th 100 53 Cr 9.501 113 Cd 12.22 In 4.3 173 Yb 16.12 233 54 Fe 5.8 Cr 2.365 114 Sn 0.65 Cd 28.73 174 Yb 31.8 Hf 0.162 234 U 0.0055 55 Mn 100 115 Sn 0.34 In 95.7 175 Lu 97.41 235 U 0.7200 56 Fe 57 Fe

91.72 2.2

116 117

Sn 14.53 Cd 7.49 Sn 7.68

176 Lu 2.59 Yb 12.7 Hf 5.206 177 Hf 18.606

236 237

58 Fe 0.28 Ni 68.077 118 Sn 24.23 178 Hf 27.297 238 U 99.2745 59 Co 100 119 Sn 8.59 179 Hf 13.629 60 Ni 26.223 120 Sn 32.59 Te 0.096 180 Ta 0.012 W 0.13 Hf 35.100

Taking Interferences into Account «on Paper»

0

500

1000

1500

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

Rela

tive

Inte

nsity

(arb

itrar

y)

m/zPd, 1 mg/kg

Cd, 50 mg/kg

0

10

20

30

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

Isot

ope

Abun

danc

e [%

]

m/zPd

Cd

Concentrationdifference

• Chose interference-free isotope• Mathematical correction

− monitor interference freeisotope of interfering species

− Calculate contribution on m/z of interest

− Subtract calculated intensityfrom analyte signal

Collision-/Reaction CellICP-MS

Adapted from: Thomas: A Beginner's Guide to ICP-MS, Part IX - Mass analyzers Collision/reaction cells

Analyte ions

Isobaric interference

Reaction gasReaction cell quadrupole

Analyzer Quadrupole

Ion-molecule-interactions- Collision- Reaction- Charge transfer

You win some, you loose someICP-MS: Reaction cell

Ar+ + NH3 Ar + NH + 3

Ca + + NH3 no reaction

Gas Flow Rate CH4 [ml/min]

Sign

al In

tens

ity[c

ps]

Adapted from B. Hattendorf, 2002, Diss ETH No. 14926

Reaction cell technology withNH3, O3, H2,... Very selective Matrix-& analyte-dependent Introduction of new

interferences possible

Collision technology with He Widely applicable for wide

range of elements Limited in sensitivity &

selectivity

Sector-field ICP-MSHigh-Resolution ICP-MS

combination of electrostatic

and magnetic sector-field

G. Fontaine, Diss. ETH No. 19483, 2010

m/z

Inte

nsity

75

Inte

nsity

m/z74.921 74.931

75As+

35Cl40Ca

+

Low resolution

High resolution

75As+

35Cl40Ca

+

ICP-MS: Characteristics# of Elements 70Data Aquisition Sequentially (fast)Sample Types Liquid, solid (after digestion), (gaseous)Limits of Detection ~0.001 μg/L (ppt-range, in solution)Dynamic Range 6 – 8 orders of magnitudeWorking Range 0.001 – 10’000 μg/L (in solution)Analytical Expertise HighCost Aquisition High (Single Quad ~100’000 €, triple Quad

~300’000 €, Sector-Field / TOF ~450’000 €)Running Cost High (Ar)

Multi-elementSensitivityGood selectivity

High demands on purityExpensiveProne to interference

57

Wrap-Up

https://milestonesci.com/wp-content/uploads/2017/07/PP_UltraCLAVE_Thumb2.png

AAS

ICP-OES

ICP-MS

Single (few) elementsComplicated matrix

Scre

enin

gs

Specification limits> 1 (10) mg/kg

Specification limits< 1 mg/kg

Scre

enin

gs

Spec

ifica

tion

for

Rel

ease

What would be a suitable sample preparation? Which considerations would be different for Tablet A and B (composition below)

Sample PreparationObtaining a liquid (uniform) sample

Solid sample (organic / inorganic)

Digest (acidic)

Open digestion

+ Direct observation, cheap, fast forsome samples

- Loss of volatile analytes /speciesformed (Hg, Os, B), acid fumes, slowfor many samples

Closed-vesseldigestion

+ Good retaining of volatile elements, high T (<300°C) & P possible(> 300°C), can be automated, lowercontamination, less acidconsumption, less residual carbon

- Memory effects in digestion vials, takes its time during methoddevelopment, high initial cost

Dissolve

In aqueoussolution(mostly

acidified)

+ Fast, low effort, low or known matrixeffects

- Only applicable to very few samples

In organicsolvent

+ Fast, well applicable to organicmatrices

- Komplex matrices (not removed), method development required

Liquid sample (organic)

Liquid sample (aqueous)

Dilute & shoot

Digestion with (Concentrated) AcidsSample Preparation

Preferred acid for dissolution, dissolves and oxidizes a wide variety of samples. Highest purity with low viscosity effects, introduces few interferences (NOx↑)Stabilization of precious metals, dissolution of metals. Cl-interferences (removable by evaporation to dryness and take up in HNO3), some analytes may be lost as volatile metal chlorides – As, Sb, Sn, Se, Ge, Hg

Not favored in ICP-MS due to high boiling point, high viscosity, interference by S-species, can be used to achieve higher temperatures during digestion

(Too) strong oxidizing power, used mainly in steel dissolution processes, can serve as alternative to HCl

Not favored (alone) since analytes tend to adhere to wall of container,acids deter this adhesion and maintain analytes in solution, can be usedfor initial dilution prior to addition of acids

• H2O

• HNO3

• HCl

• H2SO4

• HF

• HClO4

Extreme health hazard, necessary for silicate-based materials, stabilization of Ti & Hf, attacks glass/quartz, inert containers required

Adapted from D. Fontaine

Keeping analytes in solutionGotta catch them all!

• Hg: stabilizer

• PGE: Platinum group elements, Ru, Rh, Ir, Os, Pt + Au

• Ag

https://www.inorganicventures.com/periodic-table

HCl

HCl

https://www.inorganicventures.com/osmium

Compromise orstabilize separately

HNO3Digestion CxHxNx

HF

Tablet A: HNO3 + HCl or HClO4 + HF Tablet B: HNO3 + HCl / HClO4

• Typical independent external calibration in combination with internal standards• Recovery of the procedure is monitored with several standards & reference materials

QuantificationICP-MS

Sample

• Accurately weigh in sample (ca. 50 – 500 mg) and reference materials to characterize digestions procedure, prepare blank, (spike with recovery standard)

• Add acid mixture for digestion, proceed with microwave digestion procedure

Digestedsolution

• Check whether solution is clear• Fill to final weight/volume with diluent (dilution step 1, an acid concentration of 2-10% is

ideal for storage)

Dilution

• Add e.g. Y, In (Ir, Sc, Bi, Lu) for internal standardization (example)• Dilute to final weight/volume (typical dilution factors are 100-10’000, matrix should be

below 500 mg/kg)

ICP-MS

• 0.1-100 µg/kg typically for traces (in solution)• Major elements can be quantified up to 100s of mg/kg (in solution)

• Let system settle

• Run calibration

• Run calibration check solutions (SCV, recovery)

• Run digestion blanks

• Run reference materials

• Run samples

• Check stability (recovery)• Cover analyte range found

(recovery)

(ICP-MS): Measurement Sequence (Example)

• Look for patterns (which do not fit)• Get an isotope abundance table• Different concentrations obtained from different isotopes / wavelengths• Unexpectedly high concentrations e.g. for Hg (WO+?)

• Check common contaminations• Fe together with Ni and Cr steel contamination? From sample or from

preparation/analysis?• Al, B, Zn: extracted from quartz/glass? From sample container? During preparation?

• Check potential carry-over / cross contamination• Carry-Over: highly concentrated samples run directly before affected sample? Wash-out from

calibration standards?• Cross Contamination: within same run also highly concentrated samples? In the run before?

Find and identify what’s left of contamination and interferencesICP-MS: Data Evaluation

“If you are in charge of quality control laboratories in manufacturing companies, it is important to distinguish between the variability of a product and the variability of the analysis.

When analyzing tablets on a pharmaceutical production line, variability in the results of an analysis has two contributions: from the product itself and from the analytical procedure. Your bosses are interested in the former, and you, the analyst, must understand and control the latter.

It is usually desired to use methods of analysis for which the repeatability is much less than the variability of the product, in which case the measured standard deviation can be ascribed entirely to the product.“

Quality Assurance for the Analytical Chemistry Laboratory, D. Brynn Hibbert, 2007

Finding the Right Tool

Instrument Qualification

URS • User Requirement Specification

DQ • Design Qualification

IQ • Installation Qualification

OQ • Operational Qualification

PQ • Performance Qualification

SOP – Standard Operating Procedure

If your customer wanted to implement analysis of those elements for release analysis, how would you ensure that the method is “valid” to analyse them at this level – in compliance with quality standard GMP (good manufacturing practice)?

Validation of Analytical Method

SOP • Method

VP • Validation plan

✔ • Experiments (OOS, deviations)

VB • Validation report

Method Validation acc. to Ph. Eur. 2.4.20 / USP <233>Validation of Analytical Method

Parameter Solutions Acceptance CriteriaAccuracy Sample spiked 50% - 150% of

specification limit (n = 3 per level)70 – 150% recovery (per level)

Precision (Repeatability)

Sample spiked at 100% specification limit (n = 6)

RSD max. 20%

Intermediate Precision Repetition precision different analyst / day / instrument (total n = 12)

RSD max. 25%

Specificity “unequivocally assess each element in presence of matrix”Quantitation limit Shown via accuracy (50%)

Range Shown via accuracy Linearity Shown via accuracy

• Highly regulated (except for troubleshooting)• Follow the method (if available)• Use qualified instruments• Avoid contamination• Digest samples if possible to remove matrix• Choose method best suited for purpose (not necessarily most sensitive one)• Calibrate suitably• Carry out an SST• Analyse your samples• Carry out 2nd part of SST• Compare result to specification• Document your analysis• Get it reviewed

Trace Element Analysis in Pharma

Determination of minute sample volumes: Flow injection 10 – 20 µL sample material Separation of element species: Cr(IV) / Cr (VI), As(III) / As (V)

ICP-MS = DetectorOn it: Coupling HPLC-ICP-MS

Separation of element species = SpeciationCoupling HPLC-ICP-MS

Agilent Handbook: Time Resolved Analysis, Chromotographic Data Analysis, Hyphenated Technique (incl. Laser Ablation), R. Wahlen, LGC, «Fast and Accurate Determination of Arsenobetaine in Fish tissues using HPLC-ICP-MS, Agilent application note

• Different toxicity of inorganic & organic compounds• As(III)• As(V)• Anti-cancer agent As(III) in solution is oxidized to As(V)• Within the same matrix, As(III) as well as As(V) is to be analysed

Arsenic trioxideSpeciation: HPLC-ICP-MS

𝐴𝐴𝐴𝐴(𝐼𝐼𝐼𝐼𝐼𝐼)−2𝑒𝑒−

𝐴𝐴𝐴𝐴(𝑉𝑉)

Thank you

+ Jochen Bergmann & Marco Born for company slides

Happy to serve you

Now, later, just before the exam…In case questions arise:

Gisela Fontaine +41 (0) 61 845 6012 gisela.fontaine@solvias.comSolvias AGRömerpark 44303 KaiseraugstSwitzerland

(1) DH Carr, J Brown, GM Bydder, RE Steiner, HJ Weinmann, U Speck, AS Hall and IR Young, American Journal of Roentgenology. 1984;143: 215-224

(2) https://www.srf.ch/kultur/wissen/wenn-nach-der-roehre-metall-im-kopf-zurueckbleibt(3) Vecteezy.com(4) https://orgspectroscopyint.blogspot.com/2013/07/atomic-absorption-spectroscopy.html(5) S. Traxel, Merck, «Metallische Verunreinigungen», Concept Heidelberg, 06.10.2016(6) Jan Thieleke, Institute of Inorganic Chemistry (Hanover); translated by Lisa Amelung,

http://2014.igem.org/Team:Hannover/Background_ICP_OES(7) G. Fontaine, Diss. ETH No. 19483, 2010(8) http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532003000200007, Bernhard

Welz; Helmut Becker-Ross; Stefan Florek; Uwe Heitmann; Maria Goreti R. Vale, «High-resolution continuum-source atomic absorption spectrometry – What can we expect?”, J. Braz. Chem. Soc. vol.14 no.2, 2003

(9) Andrew Teasdale, Cyrille C. Chéry, Graham Cook, John Glennon, Carlos W. Lee, Laurence Harris, Nancy Lewen, Samuel Powell, Helmut Rockstroh, Laura Rutter, Lance Smallshaw, Sarah Thompson, Vicki Woodward, Katherine Ulman “Implementation of ICH Q3D Elemental Impurities Guideline: Challenges and Opportunities”, Pharm. Tech., Volume 39, Issue 3

(10) http://lab-training.com/2013/05/08/graphite-furnace-atomisation/(11) http://blogs.maryville.edu/aas/wp-content/uploads/sites/1601/2013/05/edl1-e1368818232951.gif(12) https://chem.libretexts.org/Bookshelves/Analytical_Chemistry/Book%3A_Analytical_Chemistry_2.

0_(Harvey)/10_Spectroscopic_Methods/10.1%3A_Overview_of_Spectroscopy

Most schematics in this presentation are not intellectiual propertyof the presenter but belong to the respective owners

Sources / References

(13) https://www.perkinelmer.com/CMSResources/Images/44-74440BRO_FIMS-100-400-006858D_01.pdf

(14) https://www.shsu.edu/~chm_tgc/primers/pdf/HGAAS.pdf(15) Dr. Daniel Fontaine(16) Prof. Dr. Detlef Günther(17) Dr. Bodo Hattendorf(18) H.Willard, LMerritt, J.Dean, F.Settle: Instrumental Methods of Analysis; Wadsworth Publishing

Company(19) L. Flamigni et. al. Plasma-particle interaction studied by laser scattering and optical emission

spectrometry(20) 1991 IUPAC, adapted from PerkinElmerSciex „Relative Abundance of the Natural Isotopes", 2006(21) Agilent (2005) ICP-MS Primer(22) Thomas: A Beginner's Guide to ICP-MS, Part IX - Mass analyzers Collision/reaction cells(23) Thermo ICP-MS Shortcourse(24) Bruker Shortcourse on ICP-MS(25) J. Nobrega, Federal University of São Carlos, Microwave-Assisted Sample Preparation for

Spectrochemistry – Fundamentals, Short Course during the Rio Symposium 2014(26) Agilent Handbook: Time Resolved Analysis, Chromotographic Data Analysis, Hyphenated

Technique (incl. Laser Ablation), R. Wahlen, LGC, «Fast and Accurate Determination ofArsenobetaine in Fish tissues using HPLC-ICP-MS, Agilent application note

Sources / References

(27)https://www.perkinelmer.com/lab-solutions/resources/docs/GDE_Concepts-of-ICP-OES-Booklet.pdf

(28)NZZ, 22.02.2019, Uta Neubauer https://www.nzz.ch/wissenschaft/periodensystem-der-elemente-150-jahre-ordnung-im-reich-der-chemie-ld.1456162

(29)N. Lewen et al., J. Pharm. Biomed. Anal. 35, 739–752 (2004) (30)https://www.scinexx.de/dossierartikel/meilenstein-karlsruhe/(31)https://openi.nlm.nih.gov/detailedresult.php?img=PMC4342972_rsta20140182-

g18&req=4

Sources / References

AAS Atomic absorption spectroscopy / spectrometryC(D)MO Contract (Development and) Manufacturing OrganizationCRO Contract Research OrganizationF-AAS Flame-AASGF-AAS Graphite Furnace AASETV-AAS Electrothermal Vaporisation AAS (=GF-AAS)HG-AAS Mercury-AASCV-AAS Cold vapour AAS (= HG-AAS)AES Atomic Emission spectroscopy / spectrometryGMP Good Manufacturing PracticeHPLC High performance / pressure liquid chromatographyICP-OES Inductively coupled plasma optical emission spectroscopy / spectrometryICP-AES Inductively coupled plasma atomic emission spectroscopy / spectrometry (=ICP-

OES)ICP-MS Inductively coupled plasma mass spectrometryJP Japanese PharmacopeiaPh. Eur. European PharmacopeiaUSP-NS United States Pharmacopeia – National Formulary

List of Abbreviations

Römerpark 2, 4303 Kaiseraugst, Switzerland

www.solvias.com

Gisela Fontaine, gisela.fontaine@solvias.com, T +61 845 60 12

Solvias AG