Simultaneous determination of seven major isosteroidal alkaloids in bulbs of Fritillaria by gas chromatography

The present paper describes the development of a most simple, sensitive, an d specific gas chromatographic method to date, for the direct determination of seven major bioactive isosteroidal alkaloids, namely ebeiedine, ebeiedi none, ebeienine, hupehenine, isoverticine, verticine, verticinone and imper ialine, in Fritillaria species, a commonly used antitussive traditional Chi nese medicinal (TCM) herb. In the present study, a commercially available S upelco SAG-5 capillary column (30 m x 0.25 mm, 0.25 mu m) specifically desi gned for the analysis of steroids was utilized for the direct determination of Fritillaria alkaloids. Calibration curves were obtained by spiking auth entic compounds and the internal standard (solanidine) into herbal samples prior to extraction. Extraction was conducted simply by shaking the pre-alk alized diethyl ether solution (5.0 ml) containing dried herb (0.1 g) for 2 h. All calibration curves showed good linear regressions (r(2)>0.995) withi n test ranges. The assay was reproducible and accurate with the overall int ra- and inter-day variation and accuracy of less than 10% and more than 90% , respectively. The developed GC method was successfully utilized to analyz e seven major bioactive alkaloids in seven Fritillaria species, and the res ults demonstrate that this direct GC analytical method is suitable for the quality control of this commonly used antitussive TCM herb. (C) 2000 Elsevi er Science B.V. All rights reserved.