Studies On The Quality Standards Of Cholic Acid-Verticinone Ester And Acid Percolation Method For Extracting Total Alkaloid From Fritillaria

Using the method of structural combination, a new compound named Verticinone- cholic Acid Ester was synthesized. It is a combination of cholic acid, which is one of the main active ingredients of snake's galls, and verticinone, which is one of the main active ingredient of Fritillariae. The synthetic reaction occurs between the carboxyl group of cholalic acid and the 3-hydroxy group of verticinone. By taking out pharmacological experiments, we discovered its remarkable antitussive, expectorant, antiasthmatic effects, thus making it a prospective new drμg. We carried out a preliminary study on its quality standards including physical and chemical properties.We chose HPLC-ELSD as the method for the determination after comparing it to the method of acid-dye colorimetry. Experimental conditions were as follows: Column:PurospherRSTARRP^-18e;mobile-phase:methanol:acetonitrile:triethylamine (50:50:0.1); flow rate: 1.0mL·min^-1; column temperature:25℃, detector:ELSD, Spray Chamble70℃,Drift Tube 90℃. The detection limit of Verticinone- cholic Acid Ester was 1.5μg.mL^-1, while the quantification limit was 4μg.mL^-1.The linear range was from 5.1μg.mL^-1 to 101.3μg.mL^-1, the correlation coefficient r was 0.9995. The average recovery was 98.96%. The methodology verification proves the method of HPLC-ELSD is practical in the determination of Verticinone- cholic Acid Ester.Here we carried out an orthogonal experiment on extracting total alkaloids by acid percolate methods. We compared the extraction efficiency of acid percolate methods with that of hot reflux. Acid dye colorimetry was employed for the quantitative determination. The result showed that the highest extraction efficiency occurred when: the HCl concentration was 1%, the amount was 1700 mL, the immersion time was 24 h and the percolate speed was 10 mL.min^-1·kg^-1. This method simplified the large-scale extraction of verticinone form Fritillariae hupehensis and set a foundation for accumulating the material for synthesizing Verticinone- cholic Acid Ester.