A Rapid High-Performance Liquid Chromatographic Method for the Determination of Sinapine and Sinapic Acid in Canola Seed and Meal

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1、ARapidHigh-PerformanceLiquidChromatographicMethodfortheDeterminationofSinapineandSinapicAcidinCanolaSeedandMealR.Cai1andS.D.Arntfield*DepartmentofFoodScience,UniversityofManitoba,Winnipeg,Manitoba,CanadaR3T2N2ABSTRACT:Ahigh-performanceliquidchromatographicmethods(1,14),whenusedtodetermineind

2、ividualphenolics,(HPLC)methodhasbeendevelopedtoseparatesinapineandrequirespecificreagentsforcolordevelopment.Furthermore,sinapicacidfromotherphenolicsincanolaseedandmealinaitisdifficulttoavoidtheinterferencefromothercompoundssinglerun.Theseparationwasachievedwithareverse-phaseinthesesystems.

3、SomecolorimetricmethodsandultravioletC18column.Owingtothehigherrecoveryofphenolicsand(UV)spectrophotometricmethodsrequirepurificationproce-easeofuse,refluxingwith100%methanolfor20minwasse-dures(15,16).Gaschromatography(2,17)requiresthehy-lectedastheextractionmethodforHPLCanalysisanddetermi-d

4、rolysisofphenolicestersandthederivatizationoftheresult-nationoftotalphenolicsusingFolin-Ciocalteureagent.A10-ingphenolicacidsbeforedetermination.Therefore,itisanminisocratic/linear/concavegradientanda15-minisocratic/lineargradientwereselectedasthebestgradientsforthesepa-indirectmethodandnot

5、directlyapplicabletononvolatilerationofthesephenoliccompounds.Peakidentitiesforsinap-phenolicesters.High-performanceliquidchromatographyineandsinapicacidwereverifiedwithionexchangesepara-(HPLC)isarapidandsensitivemethodforqualitativeandtionfollowedbyHPLCanalysis.Themethodwascalibratedquanti

6、tativeanalysisofplantphenolics(18).usingsinapinebisulfateandsinapicacidstandards;correlationSeveralHLPCmethodsforthedeterminationofphenoliccoefficients(R2)forthecalibrationcurveswere0.997and0.999compoundshavebeendeveloped.Clausenetal.(19)usedaforsinapinebisulfateandsinapicacid,respectively.

7、Theextinc-reverse-phaseNucleosil5C8columnandaNucleosil5C18tioncoefficientofsinapinewasdeterminedtobe1.16timesthatcolumnwitha30-minlineargradientcomposedofacetoni-ofsinapicacidatthedetectorwavelength(330nm).Applyingtrileandaphosphatebuffertoseparateagroup