PREDERIVATIZATION AND HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF ALKALOIDS OF BULBS OF FRITILLARIA

A method of prederivatization and HPLC with UV detection was developed for the simultaneous analysis of five major steroidal alkaloids of th e Fritillaria species: verticine, verticinone, isoverticine, ebeiedine , and ebeiedinone. Derivatization was carried out by esterification of the hydroxyl-containing Fritillaria alkaloids to the corresponding na phthoates with 1-naphthoyl chloride. Reaction conditions were optimize d and the yields of the derivatization were between 94 and 100% for al l test alkaloids. Derivatized alkaloids were characterized by mass spe ctrometry and HPLC-MS. The validated HPLC-UV method demonstrated linea r UV response at the sampling ranges used, and a test limit of 1 mu g/ mL was determined for all analytes. This analytical method is simple, convenient, and readily reproducible. The developed method was applied to the analysis of the major pharmacologically active alkaloids in th ree medicinally used Fritillaria species: F. cirrhosa F. thunberghii, and F. hupehensis. Five major Fritillaria alkaloids were simultaneousl y analyzed qualitatively and quantitatively from crude extracts of eac h of these herbs.