Preparative and Analytical Separation of Amygdalin and Related Compounds in Injectables and Tablets by Reversed-Phase HPLC and the Effect of Temperature on the Separation

Previous HPLC procedures for amygdalin and related compounds in injectables and tablets were either time consuming or produced inadequate separations of D-amygdalin and its epimer. A study of the effects of temperature on the separation resulted in development of an HPLC method for amygdalin and some related compounds, using water as the mobile phase at 15 degrees C. Multimilligram quantities of amygdalin and related compounds were separated by this preparative procedure. The aqueous mobile phase allows the compounds to be recovered by simple lyophilization of the sample after elution. This permitted the carbon-13 NMR spectrum of the isolated aglyconic epimer of amygdalin to be reported for the first time. D-amygdalin, its L-mandelonitrile epimer (D-epiamygdalin), their hydrolysis products (the epimeric amides and epimeric acids), and the sugar gentiobiose were separated by the method.