Established Purification And Quality Evaluation Methods Of Fritillaria Alkaloids

Bulbus Fritillariae (BF), referred to the dried bulbs of several Fritillaria species (Liliaceae), is a common traditional Chinese medicinal herb, which isregistered in the Chinese Pharmacopoeia2010edition. Chemistry and modern pharmacological studies have demonstrated that the various kinds of steroidal alkaloids are the main bioactive ingredients in Fritillaria species.Firstly, extraction procedure was optimized according to the resulting Fritillaria alkaloids’s kind and quality, and then purification method was developed.The crude alkaloids were extracted by changing pH value based on different status of alkaloids under different pH conditions. The fine alkaloids were obtained using prep-HPLC method to remove polar impurity in the crude alkaloids. Secondly, the quality evaluation method of Fritillaria alkaloids was established. The MS fingerprint method was developed to evaluate the quality of Fritillaria alkaloids and there were significant differences among six classes of Fritillaria alkaloids.Finally, the fragmentation behaviors of cevanine type and jervine type alkaloids from thunbergii and unibracteata were studied by LC-ESI-MS/MS. The cevanine type alkaloids showed predominant [M+H-H2O]+ion at50eV CE that indicated the hydroxyl substitution at C20. More fragment ions were obsersved at70eV CE.While the jervine type alkaloids showed characteristic fragmentation ion m/z67.05both at50eV and70eV CE. Based on the fragmentation pathway rules of alkaloids, combining the literatures,8alkaloid compounds from thunbergii and4alkaloid compounds from unibracteata were identified.