Micellar electrokinetic chromatography for the analysis of D-amygdalin and its epimer in apricot kernel

We have developed a simple, rapid and reproducible method for the determination of D-amygdalin and its epimer by using micellar electrokinetic chromatography (MEKC). Separation of D-amygdalin was performed in a 20 mM sodium borate buffer (pH 8.5) containing 300 mM sodium dodecyl sulfate using a bare fused-silica capillary. The eluates were monitored by the absorbance at 210 nm. The applied electric field was 278 V/cm, and the time needed for the separation of D-amygdalin did not exceed 6 min. The calibration curve for D-amygdalin showed excellent linearity in the concentration range of 5-500 microg/ml. The migration time and the corrected peak area show relative standard deviations (n=6) of 0.86% and 1.48%, respectively. The limit of detection (S/N=3) for D-amygdalin was 2 microg/ml. Under acidic and neutral conditions, amygdalin exists only as the D-form; however, under basic conditions, it shows both the D- and L-forms with a concentration ratio of 1:1.3 (D-amygdalin/L-amygdalin). Results of HPLC, UV-Vis spectrophotometry, and mass spectrometry reconfirmed the identification of D-amygdalin and its epimer. The number of theoretical plates of D-amygdalin is about 100,000 in MEKC, which is significantly higher than approximately 8,000 of HPLC. This method has been successfully applied to the determination of amygdalin epimers in various apricot kernel extracts and pharmaceutical products.