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1、FCCVMonographs/Biotin/49andrecordtheareasasdirectedunderProceduretoensureREQUIREMENTSthattherelativestandarddeviationdoesnotexceed2.0%forIdentificationDissolve200mgof3,3′-dimethoxybenzidinethe3-tert-butyl-4-hydroxyanisoleisomerand6.0%forthe2dihydrochlorideinamixtureof40
2、mLofmethanoland60tert-butyl-4-hydroxyanisoleisomer;theresolutionbetweenmLof1Nhydrochloricacid.Add5mLoftheresultingtheisomersisnotlessthan1.3;andthetailingfactordoesdianisidinesolutionto10mLofwaterand2mLofa3:100notexceed2.0.solutionofsodiumnitrite.Addthissolutionto10mLof
3、aProcedureSeparatelyinjectsuitableportions(about51:100,000solutionofsample:methanol.Anorange-redcolor�L)oftheStandardPreparationandtheAssayPreparationappearswithin3min.Add5mLofchloroform,andshake.intothegaschromatograph,andrecordthechromatograms.Thechloroformlayerexhibi
4、tsapurpleormagentacolorthatMeasuretheareasunderthepeaksforeachisomerandthefadeswhenexposedtolight.InternalStandardineachchromatogram,andcalculatetheAssayNotlessthan99.0%ofC15H24O.quantity,(I),inmilligrams,ofeachisomerinthesampletakenResidueonIgnitionNotmorethan0.002%.by
5、theequationI=10×CS×(RU/RS),TESTSinwhichCSistheconcentration,inmilligramspermilliliter,AssayThesolidificationpoint(seeSolidificationPoint,ApoftheisomerintheStandardPreparation;RUistheratioofpendixIIB)ofasampleisnotlowerthan69.2°,indicatingatheareaoftheisomertothatoftheIn
6、ternalStandardinthepurityofnotlessthan99.0%ofC15H24O.chromatogramfromtheAssayPreparation;andRSistheratioResidueonIgnitionTransferabout50gofsample,accuoftheareaoftheisomertothatoftheInternalStandardinratelyweighed,intoataredcrucible,igniteuntilthoroughlythechromatogramfr
7、omtheStandardPreparation.Calculatecharred,andcool.Moistentheashwith1mLofsulfuricacid,theweight,inmilligrams,ofC11H16O2inthesampletakenandcompletetheignitionbyheatingfor15-minperiodsatbyaddingthequantitiesofthetwoisomers.800°�25°toconstantweight.ResidueonIgnitionDetermin
8、easdirectedinMethodIunderResidueonIgnition,AppendixIIC,ignitinga10-gPackagingandStorageStoreinwell-closedconta
