additive-467-m1 dl- α -Tocopherol(Vitamin E; INS N

dl-α-TOCOPHEROLPreparedatthe30thJECFA(1986),publishedinFNP37(1986)andinFNP52(1992).Metalsandarsenicspecificationsrevisedatthe61stJECFA(2003).AgroupADIof0.15-2mg/kgbwfordl-α-tocopherolandd-α-tocopherol,concentrate,singlyorincombination,wasestablishedatthe30thJECFA(1986).SYNONYMSVitaminE;INSNo.307cDEFINITIONChemicalnamesdl-5,7,8-Trimethyltocol,dl-2,5,7,8-tetramethyl-2-(4',8',12'-trimethyltridecyl)-6-chromanolC.A.S.numberNosingledefiniteC.A.S.numberisforthissubstance.No.59-02-9isforvitaminE,1406-18-4isforalpha-tocopherol,2074-53-5isforall-rac-alpha-tocopherol,and10191-40-0isforracemic-alpha-tocopherolsynthesizedfromnaturalphytoloritsderivative.ChemicalformulaC29H50O2StructuralformulaFormulaweight430.71AssayNotlessthan96%andnotmorethan102%DESCRIPTIONSlightlyyellowtoamber,nearlyodourless,clear,viscousoil(Itoxidizesanddarkensinairandonexposuretolight.)FUNCTIONALUSESAntioxidantCHARACTERISTICSIDENTIFICATIONSolubility(Vol.4)Insolubleinwater,freelysolubleinethanol,misciblewithetherSpecificrotation(Vol.4)[alpha]20,D:0±0.05o(1in10solninchloroform)Spectrophotometry(Vol.4)Inabsoluteethanolthemaximumabsorptionisabout292nmColourreactionDissolveabout0.01gofthesamplein10mlofabsoluteethanol.Add,withswirling,2mlofnitricacidandheatatabout75ofor15min.AbrightredtoorangecolourdevelopsPURITYRefractiveindexn(20,D):1.503-1.507Spectrophotometry(Vol.4)E(1%,1cm)(292nm):71-76(0.01gin200mlofabsoluteethanol)Sulfatedash(Vol.4)Notmorethan0.1%Test1gofthesample(MethodII)AcidityDissolve1gofthesamplein25mlofamixtureofequalvolumesofethanolandetherthathasbeenneutralizedtophenolphthaleinTSwith0.1Nsodiumhydroxide,add0.5mlofphenolphthaleinTS,andtitratewith0.1Nsodiumhydroxideuntilthesolutionremainsfaintlypinkaftershakingfor30sec.Notmorethan1.0mlof0.1Nsodiumhydroxideisrequired.Lead(Vol.4)Notmorethan2mg/kgDetermineusinganatomicabsorptiontechniqueappropriatetothespecifiedlevel.TheselectionofsamplesizeandmethodofsamplepreparationmaybebasedontheprinciplesofthemethoddescribedinVolume4,“InstrumentalMethods.”METHODOFASSAYGasLiquidChromatographicMethodReagentsandsolutionsInternalStandardSolution:Prepareasolutioninn-hexanecontaininga3mgofhexadecylhexadecanoate,accuratelyweighed,ineachml.StandardPreparation:dissolveabout30mgofUSPAlphaTocopherolReferenceStandard,accuratelyweighed,in10.0mloftheInternalStandardSolution.AssayPreparation:dissolveabout30gofthesampleaccuratelyweighed,in10.0mloftheInternationalStandardSolution.ChromatographicSystemUseagaschromatographequippedwithaflame-ionizationdetectorandaglass-linedsample-introductionsystemoron-columninjection.Undertypicalconditions,theinstrumentcontainsa2-mx4-mmboro-silicateglasscolumnpackedwith2%methylpolysiloxanegumon80-to100-meshacid-basewashedsilinizedchromatographicdiatomaceousearth.Thecolumnismaintainedisothermallybetween240oand260o,theinjectionportatabout290o,andthedetectorblockatabout300o.Theflowrateofdrycarriergasisadjustedtoobtainahexadecylhexadecanoatepeakapproximately18to20minaftersampleintroductionwhena2%columnisused,or30to32minwhena5%columnisused.(NOTE:Cureandconditionthecolumnasnecessary).SystemSuitabilityChromatographasufficientnumberofinjectionsofamixtureinn-hexaneof1mgpermleachofUSPAlphaTocopherolReferenceStandardandUSPAlphaTocopherylAcetateReferenceStandard,asdirectedunderCalibration,toassurethattheresolutionfactor,R,isnotlessthan1.0.CalibrationChromatographsuccessive2-to5-µlportionsoftheStandardPreparationuntiltherelativeresponsefactor,F,isconstant(i.e.,withinarangeofapproximately2%)forthreeconsecutiveinjections.Ifgraphicintegrationisused,adjusttheinstrumenttoobtainatleast70%maximumrecorderresponseforthehexadecylhexadecanoatepeak.Measuretheareasunderthemajorpeaksoccurringatrelativeretentiontimesofapproximately0.51(alpha-tocopherol)and1.00(hexadecylhexadecanoate),andrecordthevaluesasAsandA1,respectively.Calculatetherelativeresponsefactor,F,bytheformula(As/A1)x(C1/Cs),inwhichC1andCsaretheexactconcentrations,inmgperml,ofhexadecylhexadecanoateandofUSPAlphaTocopherolReferenceStandardintheStandardPreparation,respectively.ProcedureInjectasuitableportion(2to5µl)oftheAssayPreparationintothechromatograph,andrecordthechromatogram.Measuretheareasunderthemajorpeaksoccurringatrelativeretentiontimesofapproximately0.51(alpha-tocopherol)and1.00(hexadecylhexadecanoate),andrecordthevaluesasauanda1,respectively.Calculatetheweight,inmg,ofdl-alpha-tocopherolinthesamplebytheformula(10C1/F)x(au/a1).