Simultaneous Determination of L-Ascorbic Acid and L-Ascorbylpalmitate Using RP-HPLC Method

Abstract

A rapid reversed phase highperformance liquid chromatography (RP-HPLC) method was developed for the simultaneous analysis of L-ascorbic acid and L-ascorbylpalmitate (AP). The chromatography was performed on a SSI model 2300-525 high performance liquid chromatographequipped with anAlltechApollo C18 column at 30 oC.The mobile phase was acetonitrile-water (90:10,v/v) with the flow rate of 1.0 mL/min. UV detection wavelength was 250 nm. This method permits the simultaneous determination of ascorbic acid and AP in the synthesis of AP transesterified with methyl palmitate and ascorbic acid. The detection limit of ascorbic acid and AP was 0.07μg/mL and 0.12μg/mL, respectively. The recovery was 90.59 ± 3.04% for ascorbic acid and 101.3 ± 4.81% for AP. The linearity range for ascorbic acid and AP was in the range of 0.1 - 0.7 mg/mL and 0.4 - 4.0 mg/mL, respectively. Correlation coefficients (R2) were 0.9910 for ascorbic acid and 0.9986 for AP. The proposed method could be used for routine quality control of AP synthesis with methyl palmitate and ascorbic acidby enzymatic transesterification.