Dr. Gisela Fontaine Teamleader ICP-MS, Solvias AG 01.10.2019 Inorganic Trace Element Analysis in the Pharmaceutical Industry
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 [email protected] 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
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www.solvias.com
Gisela Fontaine, [email protected], T +61 845 60 12
Solvias AG