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


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





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





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





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





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





BrightSpec Thermal Evolution Method-Heatdrypowderinanevacuatedheadspacevial,followedbyvacuum-driventransferof

headspacevaporintoFT-MRRchamberforcharacterization

-Canperformbroadbandinvestigativeanalysis(includingunknowns)ortargetedanalysisofknowncompounds

-Athighertemperatures,degradationproductscanbegeneratedaswellasimpurities(thoughoxygen-freeenvironmentpreventscombustion)

-Verysensitivefordrypowdersbecausevolatileimpuritiescanbeseparatedefficientlyfrommatrix





Profile Results

Chloroethane resolvedincomplexmix





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)





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





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





Analysis Methods1)CharacterizationofFT-MRRsignaturesoftargetanalytesusingbroadbandspectrometer

ConfirmedidentityofDHAAbymeasuringfrequencyshiftsupon13Cisotopicsubstitution– detectedateachpositioninnaturalabundance.(12hourmeasurementtime,~100mgsample)

(Whenlesssampleisavailable,comparisonsbetweenexperimentalandtheoreticalparametersaresufficientforID.)

Data:BrooksPate,Univ.ofVa.

DHAAcomputed3-Dstructurewithexperimentalcarbonpositions(smallcircles)

DHAA(2conformersassigned)

Undesiredproducts(2conformersassigned)

LineFWHM70kHz





Analysis Methods2)Cavity-enhancedspectrometercoupledtosamplingmanifoldforrapidanalysis

Prototypedesign,basedonspectrometerdesignofR.D.Suenram etal.

Currentcycletime:18minuteslimitedbyspeedofthermalcycling,evacuationofreservoirbetweensamples

Firsttestcase:conformerratioinpureDHAA(setbybeamdynamics)

C1 C2

Connectstocontinuousmanufacturingsamplelinedirectly– duetosamplehandlingrequirementsdoesnottakeinsamplecontinuously Currentlycandetectdownto5%impurity,with

workinprogresstoextenddownto<1%





Summary

FT-MRRisbeingappliedtoarangeofproblemsinimpuritycharacterization–bothvolatileresidualimpuritiesandchiralstructuralanalysis(differentinstruments,sametechnique)

Advantagesoverothertechniquesincludesimplermethoddevelopmentandoperation,straightforwardresolutionofcomplexmixtures- includingisomers,andsimplequantitation

Speedandeaseofmeasurement,linearityofresponse,andquantitativeaccuracyareallontracktomeetcustomerexpectations

WeareseekingcollaborationsondevelopingapplicationsofFT-MRRinprocessR&Denvironments!


Thank YouFinancialSupport

Contactus:Science: JustinNeill,CTO

[email protected]:BobLloyd,CEO

[email protected]

770HarrisSt,Suite104bCharlottesville,VA22902+1(434)202-2391

http://www.brightspec.com

Virginia BiosciencesHealth ResearchCorporation

applications of ft-mrr spectroscopy for impurity identification...

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