Applications of FT-MRR Spectroscopy for Impurity Identification...
Transcript of Applications of FT-MRR Spectroscopy for Impurity Identification...
Molecular Rotational Resonance Spectroscopy
BrightSpec,Inc.|770HarrisSt.#104b|Charlottesville,VA22903|(434)202-2391
©2017BrightSpec,Inc.
Applications of FT-MRR Spectroscopy for Impurity Identification and
Quantification
Justin L. Neill, PhDChief Technology Officer
BrightSpec, Inc.
2017CPACSpringMeetingUniversityofWashingtonMay1,2017
CPCPACSpringMeetingMay1,2017
FT-MRR BasicsFourierTransform–MolecularRotationalResonance
§ Moleculeshavefingerprintrotationalspectrabasedon3-dimensionalmomentsofinertia– isomers,conformers,andisotopologues resolved(Additionalcapabilitiesforenantiomers)
§ Extremelyselective– nofalsepositives
§ SpectrameasuredusingFTtechnique(analogoustoFT-NMR)thatdramaticallyenhancessensitivity
§ Analytetargetsare:§ Volatilizable – measurementisinlowpressuregas
phase§ Polar – interactionisthroughpermanentdipole
moment§ Lowweight– <125amu forroom-temperature
analysis,upto500amu usingmolecularbeams
Simplerelationshipbetweenabinitioelectronicstructureandspectra
Site-specificisotoperesolution
CPCPACSpringMeetingMay1,2017
BrightSpec• Foundedin2012tocommercializeinnovative
technologyfromtheUniversityofVirginia.IPportfolioof7patents(UVa,Harvard,andBrightSpec-filed)
• Teamof10full-timeemployees- 4expertsinFT-MRRtechnique- Complementaryexpertiseinengineering,applicationsdesign,instrumentdesign,software- Experiencedmanagementteam
• BasedinCharlottesville,VA
BrightSpecFT-MRROne BrightSpecDiscoverySeries BrightSpecChiralBroadband
AllproductsCEcertified21CFRPart11compatible
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
OutlineIwillpresentthreeexamplesofclient-drivenanalyseswhereFT-MRRisbeingapplied:
1) QuantificationofresidualsolventimpuritiesinnutritionalIVsolutions,rawmaterials,anddrugproducts
2)ID&quantificationofatracelevelmutagenicimpurityinadrugproduct
3) Chiralpuritymonitoringofacontinuouspharmaceuticalsyntheticprocess
CollaborationwithB.FrankGupton,VirginiaCommonwealthUniversity
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Residual Solvent Analysis by FT-MRRResidualsolventanalysis(USP<467>):
- Pharmaceuticalmanufacturersmustverifythatresidualimpuritiesusedinsynthesisareatsafelevelsintheirproducts– typicallyppmsensitivityrequired(Class2)
- Gaschromatographyisstandard,butsimplerandfastermethodsaredesirableiftheycandemonstrateequivalentperformance
ClientsentsamplesofaIV-administerednutritivesolutionwithimpuritiesofmethanol,ethanol,andisopropanoltheyneededtoquantify.Samplesarealsothermallyunstable- aproblemforGC.
FT-MRRRequirements:- Unambiguouslyresolveanalyteswithoutchromatography- Reachdetectionlimitsof1mg/Lwith<10%measurementaccuracy- Simpler,faster,andeasieranalysisthanexistingmethods
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Analytical MethodsStaticHeadspaceFT-MRR
• SimilaradvantagesasSHS-GC(concentrationofvolatiles;matrixsimplification)
• Onlyconsumableisnitrogenforcleaning
• Interfaceswithautosampler
-Referencelibraryusedtofindlinesofeachanalytewithoutoverlaps
-Methodvalidityestablishedbymeasuringstocksolutionsinconcentrationrange1-100mg/L
-Regularblankstoconfirmnocarryover-Accuracymaintainedincustomer’ssamplematrix–
demonstratedthroughspikedrecoveryanalysis
Description Time
Evacuatevial 2min
Injectsolutionandequilibrate 3min
Transfersampletomeasurementchamber
45sec
Measureanalytes 15sec(5sec/analyte)
Cleansystem 3min
Total 9min
FT-MRRMeasurementCycle
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Method Results
R²=0.9976R²=0.9960R²=0.9941
0
0.0005
0.001
0.0015
0.002
0.0025
0 20 40 60 80 100
Signal(m
V)
Concentra2on(mg/L)or(ppm)
Intensity(mV)vs[Methanol](ppm)
Trial1Trial2Trial3
R²=0.9937
0
0.0005
0.001
0.0015
0.002
0.0025
10 30 50 70 90 110
Signal(m
V)
Concentra2on(mg/L)or(ppm)
Intensity(mV)vs[Methanol](ppm)
Alltrials
0
0.0002
0.0004
Inte
nsity
(mV
)
Isopropanol Signal in Sample (279762.1 MHz)
Trial 1Trial 2Trial 3Trial 4Trial 5279762.1 MHz
279750 279760 2797700
0.0001
Frequency (MHz)
Water Blanks
LDL = 1 mg/L
MDL:Methanol 0.2mg/LIsopropanol 1.0mg/LEthanol 0.3mg/L
Repeatability typ 10%
Linearity R2 >0.99
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Mutagenic Impurity Analysis-PoliciesoutlinedinICH-M7fortheassessmentandcontrolofimpuritiesthatare
potentiallyDNAreactive
-Canarisefromrawmaterialimpurities,sidereactions,ordegradation– andmaybedescribedasasetofrelatedstructures,sostructurecharacterizationmaybeneededaswellasquantification
-Lowdetectionlimits(<1ppm)needed,withgoodquantification– challengeformethoddevelopment
Client(Top25globalpharma)providedformulateddrugcapsuleswithaknownchloroethaneimpurity.Goalistoquantifychloroethane,aswellastolearnotherinformationaboutthesamplesusingFT-MRR.
FT-MRRRequirements:-StructureIDandquantificationcapabilitiesinonemeasurement-Lowdetectionlimits-Goodreproducibilityandaccuracy
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
BrightSpec Thermal Evolution Method-Heatdrypowderinanevacuatedheadspacevial,followedbyvacuum-driventransferof
headspacevaporintoFT-MRRchamberforcharacterization
-Canperformbroadbandinvestigativeanalysis(includingunknowns)ortargetedanalysisofknowncompounds
-Athighertemperatures,degradationproductscanbegeneratedaswellasimpurities(thoughoxygen-freeenvironmentpreventscombustion)
-Verysensitivefordrypowdersbecausevolatileimpuritiescanbeseparatedefficientlyfrommatrix
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Profile Results
Chloroethane resolvedincomplexmix
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Quantitative ResultsBrightSpecsoftwaredirectlydeterminesanalytepartialpressuresinmeasurementchamber,whichareconvertedtoimpuritymassinoriginalsample.
Chloroethanemass
(w/w,ppm)
RSD(n=3)
LotA 8.8 11%
LotB 8.4 12%
LotC 14.0 3%
MDL <0.1
Chloroethane/N2 standard,partialpress.13µTorr
3samplesfromlotC
(propionaldehyde)
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Chiral Analysis in Continuous Manufacturing-Interestincontinuousmanufacturingofpharmaceuticalsisgrowingrapidly
-InAPIsynthesis,chiralityisacriticalattributethatisverychallengingtomeasure– withnogoodmethodforautomated,on-lineanalysis
-FT-MRRisuniqueinthatevenverysubtlestructuralchangescauseverycleardifferencesinthespectrum
-Indrugswithmultiplechiralcenters,unwantedsideproductsmaybediastereomersofthemainproduct– givingdifferentfingerprintsinFT-MRR
-Inthelastfewyears,twomethodshavebeendevelopedtoresolveenantiomersbyFT-MRR:1)3-wavemixing(D.PattersonandJ.Doyle,Harvard) – enantiomersproduce
radiationoppositeinphaseunderparticularconditions2)Chiraltagging(B.H.Pate,Univ.ofVa.) – sampleiscomplexedwithachiralresolving
agent,producingstructurallydistinctspecies
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Artemisinin-Wellknownantimalarial(discoveredbyTuYouyou,2015NobelPrizeinMedicine)
NaturalproductoriginallyisolatedfromaformofwormwoodnativetoAsia,butnowchemicallysynthesized
ArtemisinincombinationtherapyrecommendedfortreatmentofP.falciparummalaria
B.F.Gupton,Va.CommonwealthUniversity
Virginia BiosciencesHealth ResearchCorporation
Artemisinic Acid(Madebygenetically
modifiedyeast)
DihydroartemisinicAcid
Firsttarget:Hydrogenationstepcreatesa5th chiralcenterandHPLC+NMRarecurrentlyusedoff-linetoassessthereactionspecificityafterthefact.Anon-linesolutionisdesired.
Artemisinin
Ru/Ccatalyzedhydrogenation
1)O2/hn2)Acid-cat.
cyclization
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Analysis Methods1)CharacterizationofFT-MRRsignaturesoftargetanalytesusingbroadbandspectrometer
ConfirmedidentityofDHAAbymeasuringfrequencyshiftsupon13Cisotopicsubstitution– detectedateachpositioninnaturalabundance.(12hourmeasurementtime,~100mgsample)
(Whenlesssampleisavailable,comparisonsbetweenexperimentalandtheoreticalparametersaresufficientforID.)
Data:BrooksPate,Univ.ofVa.
DHAAcomputed3-Dstructurewithexperimentalcarbonpositions(smallcircles)
DHAA(2conformersassigned)
Undesiredproducts(2conformersassigned)
LineFWHM70kHz
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Analysis Methods2)Cavity-enhancedspectrometercoupledtosamplingmanifoldforrapidanalysis
Prototypedesign,basedonspectrometerdesignofR.D.Suenram etal.
Currentcycletime:18minuteslimitedbyspeedofthermalcycling,evacuationofreservoirbetweensamples
Firsttestcase:conformerratioinpureDHAA(setbybeamdynamics)
C1 C2
Connectstocontinuousmanufacturingsamplelinedirectly– duetosamplehandlingrequirementsdoesnottakeinsamplecontinuously Currentlycandetectdownto5%impurity,with
workinprogresstoextenddownto<1%
CaseStudy1:Routineanalysisofresidualsolventimpurities
CaseStudy2:Genotoxicimpuritiesinafinaldrugproduct
CaseStudy3:Chiralanalysisduringcontinuousmanufacturingprocess
CPACSpringMeetingMay1,2017
Summary
FT-MRRisbeingappliedtoarangeofproblemsinimpuritycharacterization–bothvolatileresidualimpuritiesandchiralstructuralanalysis(differentinstruments,sametechnique)
Advantagesoverothertechniquesincludesimplermethoddevelopmentandoperation,straightforwardresolutionofcomplexmixtures- includingisomers,andsimplequantitation
Speedandeaseofmeasurement,linearityofresponse,andquantitativeaccuracyareallontracktomeetcustomerexpectations
WeareseekingcollaborationsondevelopingapplicationsofFT-MRRinprocessR&Denvironments!
CPCPACSpringMeetingMay1,2017
Thank YouFinancialSupport
Contactus:Science: JustinNeill,CTO
[email protected]:BobLloyd,CEO
770HarrisSt,Suite104bCharlottesville,VA22902+1(434)202-2391
http://www.brightspec.com
Virginia BiosciencesHealth ResearchCorporation