Detection and identification of Extractables and leachables from Pharmaceutical Products using LC-MS

Detection and identification of potential bioactiveExtractables and leachables from Pharmaceutical Products by using LC-MS analytical technique.

1. Detection & identification of
Potential Bioactive Leachables and
Extractables from Pharmaceutical
Products By Using LC-MS
Presented By: Ashitosh Panchal
M Pharm Sem-I
Pharmaceutical Chemistry
Dr. D.Y. Patil College Of Pharmacy Akurdi, Pune

2. Contents
•What are Extractables and Leachables?
•Why do we needs E & Ls Testing?
•Guidelines
•Confident identification
•Different analytical techniques
•Conclusion
•References

3. Introduction
Extractables:
These are chemical compounds and inorganic elements
that can be forced to migrate from the contact material
(container, syringe, tubing etc.) under aggressive
conditions such as extreme surface exposure, elevated
temperature and/or strong solvents.
Leachables:
These are chemical compounds and inorganic elements
that migrate from the contact material into the product
of concern under normal conditions of use. Leachables
are usually a subset of extractables.

4. Who Needs E&L Study?
Biomedical devicesFood PackagingPharmaceutical Packaging

5. PlasticMaterials: Source of Contamination
Sources of extractables are plastic and elastomeric components
(monomers, polymeric initiators, plastizicers, etc.) ink and adhesives
(label) and degradation products (processing, storage, sterilization)
GLASS
₋ If glass is coated (with silicone),
a study may need to look
specifically for extractables that
are coming from coating. Needs
to be determined based on the
drug product formulation and
the potential for it to interact
with the coating.

6. Harmful Compounds Identified as
Extractable/Leachables:

7. Case Study Extractables and leachables:
In2010,Kellogg’sproductrecallwaslaunchedfor28millionboxesofbreakfastcerealafter
consumersreportedastrangetasteandodour,withsomecomplainingofnauseaand
diarrhoea.Thesesymptomsweresubsequentlyfoundtobecausedbyelevatedlevelsof
methylnaphthaleneintheproductpackaging.
Alsoin2010,aproductrecallwaslaunchedforamedicalproduct,Tylenol(paracetamol),
usedforpainreliefandthereliefofcoldandflusymptoms,afterconsumerreportingofa
strangeodourcomingfromthebottlesinwhichtheproductwasstored.Theodourwas
laterlinkedto2,4,6-tribromophenol,awoodpreservative,thatwaspresentwithinwooden
palletsthatwereusedtotransfertheproduct.
Theseexamplesandmanylikethemthereforehighlighttheneedtounderstandpotential
sourcesofleachablesandtoproactivelylookforandcontrolthem.

8. Risk-Based Approach To Evaluating E&L
Safety Considerations
•Toxicity, immunogenicity etc.
Efficacy considerations
•Leachables interacting with a product
•Loss of activity
•Leachable may induce development of neutralising activity
Quality considerations
•Impact on manufacturing process, product stability etc

10. Guidelines:
•1993 CDRH -Reviewer Guidance for Nebulizers, Metered Dose Inhalers,
Spacers and Actuators
•1998 FDA -MDI/DPI Draft Guidance
•1999 FDA -Guidance for Industry: Container Closure Systems for Packaging
Human Drugs and Biologics
•2002 FDA –Guidance on Inhalation solution, suspension, spray and nasal
spray products
•2005 CHMP, CVMP -Guideline for Plastic Immediate Packaging Materials
•2006 PQRI –Safety Thresholds & Best Practices For Extractables &
Leachables

11. ICH Guidelines:
•ICH Q3A: Chemical Impurities in new drug substances
•ICH Q3B: Impurities in new drug products
•ICH Q3C: Impurities: Residual solvents
ICH Q3E: Guideline for Extractables and Leachables (E&L)
•ICH Q5E: Comparability of biotechnology/biological products subject to
changes in their manufacturing process 2005
•ICH Q7A: GMP of active pharmaceutical ingredients
•ICH Q8: Pharmaceutical Development 2006
•ICH Q9: Quality Risk Management 2006

12. The Analytical Techniques For E & L Analysis

13. Liquid Chromatography-Mass Spectrometry
•Liquidchromatography-Massspectrometry(LC-MS)isapowerful
analyticaltechniquethatcombinestheresolvingpowerofliquid
chromatographywiththedetectionspecificityofmassspectrometry.
•Liquidchromatography(LC)seperatesthesamplecomponentsandthen
introducestothemassspectrometer(MS).TheMScreatesanddetects
chargedions.
•TheLC-MSdatausedtoprovideinformationaboutthemolecularweight,
structure,identityandquantityofspecificsamplecomponents.
•InLC-MSweareremovingthedetectorfromthecolumnofLC-MSand
fittingthecolumntointerfaceofMS.
•InmostofthecasestheinterfaceusedinLC-MSareionizationsource.

14. High Pressure Liquid Chromatography (HPLC)
•LiquidChromatographygenerallyutilizesverysmallparticlespacked
andoperatingatrelativelyhighpressure,andisreferredtoashigh
performanceliquidchromatography.
•ModernLC-MSmethodsuseHPLCinstrumentationessentiallyfor
sampleintroduction.
•InHPLC,thesampleisforcedbyaliquidathighpressure(themobile
phase)throughacolumnthatispackedwithastationaryphase
generallycomposedofirregularlyorsphericallyshapedparticles
chosenorderivatizedtoaccomplishparticulartypesofseparations.

15. Mass Spectrometry
•MassSpectrometry(MS)isananalyticaltechniquethatmeasuresthe
mass-to-chargeratioofchargedparticles.
•MSworksbyionizingchemicalcompoundstogeneratecharged
moleculesfragmentsandmeasuringtheirmasstochargeratio.
•InatypicalMSprocedure,asampleisloadedontotheMSinstrument
andundergoesvaporization.Thecomponentsofthesampleare
ionizedbyoneofavarietyofmethods(eg.Byimpactingthemwithan
electronbeam)whichresultsintheformationofchargedparticles
ions.Theionsareseparatedaccordingtotheirmasstochargeratioin
ananalyserbyelectromagneticfields.

16. Instrumentation of LC-MS
1.HPLC Constitutes the LC Part:
a)Solvent System ( Mobile Phase )
b)Pumps
c)Mixer
d)Injector
e)Column
2. Mass Spectrometer
A)Ion sources
i) Electrospray ionization
ii) Atmospheric pressure
Chemical ionization
iii) Photoionization

17. B) Mass Analyzers
1.Quadrupole
2.Time-of-flight
3.Ion Trap
4.Fourier transform-ion cyclotron resonance ( FT-ICR )

18. Sample Preparation
Sampleextractswerepreparedbycuttingpackagingmaterialsinto5
mm2piecesandsonicating1gofthepiecesin10mLof2-propanolfor
ninehoursatanaveragetemperatureof45°C.

19. LC Conditions
LC system: ACQUITY UHPLC I-Class FTN
Column(s): CORTECS C18 Column, 90Å, 1.6 µm,2.1 mm X 100 mm
(p/n: 186007095)
Column temp.: 40 °C
Injection volume: 1 µL
Flow rate: 0.4 mL/min
Mobile phase A: Water + 1 mM ammonium acetate +0.1% acetic acid
Mobile phase B: Methanol

20. MS Conditions
MS system: Vion IMS QTof
Ionization mode: ESI+, ESI-
Acquisition mode: HDMSE
Acquisition range: 50–1500 m/z
Scan time: 0.2sec
Source temp.:120°C
Desolvationtemp.: 500 °C
Desolvation gas: 1000 L/hr
Cone gas: 50 L/hr
Reference mass: Leucine enkephalin [M+H]+ m/z556.276

23. •LC-MS identification of E&Ls

24. Conclusion
•ThestudyhasbeenshownthattheLC-MSisthevaluabletool
fordetectionandidentificationofextractablesandleachables.
•Properlyconductedcontrolledextractablesandleachables
studies,whenaccomplishedearlyinthepharmaceutical
developmentprocess,permitapharmaceuticaldevelopment
teamtobeginearlyevaluationofpotentialdrugproduct
leachables.Thisevaluationcanalertthepharmaceutical
developmentteamtopotentialleachableswithtoxicological
concern,allowingadequatetimetobeginappropriatesafety
qualificationstudies.

25. References:
1.https://www.agilent.com/cs/library/eseminars/Public/Analysis_of_
Extractables_Leachable_Compounds_in_Pharmaceutical_Products
2.www.wikipedia.com_extractables_&_leachables
3.USFDA,CodeofFederalRegulations,21CFR211.94http://www.accessd
ata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=211.94
4.Guidance for industry: Container closure systems for packaging
humandrugsandbiologicshttp://www.fda.gov/downloads/Drugs/Gui
dances/ucm070551.pdf
5.https://www.pall.com/content/dam/pall/biopharm/lit-library/non-
gated/scientificandtechnicalreports/12.5361_USTR2859_Extractabl
es_Leachables_Single-Use_Systems_TR_EN.pdf