Determination of Ceramide in Konjac Flying Powder by Precolumn Derivation-High Performance Liquid Chromatography
An analytical method for the determination of ceramide in konjac flying powder by precolumn derivatization-high performance liquid chromatography(HPLC)was developed.Ceramide was deacylated with KOH ethanol solution at high temperature for a period of time,and then reacted with 9-fluorenylmethyl chloroformate(FMOC-Cl).The optimum conditions for deacylation and derivatization were determined as follows:Ceramide was treated with 0.5 mol/L KOH ethanol solution at 110 ℃ for 1.5h;then derived online by 10 mg/mL FOMC-C1.The separation was performed with Waters Symmetry C18 column(250 mm×4.6 mm,5 μm).Water/methanol/phosphoric acid(90/10/0.05,v/v/v)-methanol was used as mobile phase for gradient elution.The linear range of ceramide was 20-500 μg/ml(UV R2=0.9978;FLD R2=0.9978),and the average recovery was 97.3％-103.6％with relative standard deviations(RSD)less than 1.64％.The limits of detection(LOD)and quantitation(LOQ)were 0.48μg/mL(UV)and 0.0082 μg/mL(FLD),and 1.6 μg/mL(UV)and 0.027 μg/mL(FLD).
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