ABSTRACT
Objective: To develop a simple and accurate HPLC-DAD method for simultaneous determination, the content of major components: limonin, evodiamine, and rutaecarpine in Evodiae fructus and evaluation the quality of Evodiae fructus sold in markets. Methods: Open column chromatography was used to separate and purify rutaecarpine and evodiamine, the two major alkaloids from Evodiae fructus extract as a laboratory standard. Chromatographic separation was achieved using a Germini C18 column (150 mm × 4.6 mm I.D., 5 µm), detected at 210 nm. The mobile phase consisted of acetonitrile (A), methanol (B), and water (C). The validated method simultaneously determined alkaloid content in 40 batches of samples collected from markets in different regions of Vietnam. Results: In one-step purification, our method yielded 326 mg of rutaecarpine and 128 mg of evodiamine from 3.2 g of crude extract, with purities of 98.9 and 98.5%, respectively. The structures of these compounds were identified using 1H NMR and 13C NMR. There was a significant correlation between alkaloid content and fruit size, with a Spearman correlation coefficient of>0.5 (p<0.001), and there was a large difference in alkaloid contents between three maturity degrees of the fruit. Open-mouth fruits and fruits with average sizes of 4 to 6 mm had the highest alkaloid contents, whereas closed-mouth fruits had the lowest. Conclusion: This study provided information on the standardization and quality control of evodiamine and rutaecarpine in Evodiae fructus, as well as a foundation for further pharmacological and toxicological studies.
ABSTRACT
The molluscicidal activity of Hammada scoparia leaf extracts and the principal alkaloids isolated from them (carnegine and N-methylisosalsoline) were tested against the mollusc gastropod, Galba truncatula, the intermediate host of Fasciola hepatica in Tunisia. The results indicated that the molluscicidal activity was correlated with the presence of alkaloids. A significant molluscicidal value, according to the World Health Organization, was found with the methanol extract (LC50 = 28.93 ppm). Further fractionation of the methanolic extract led to the isolation of two principal alkaloids: carnegine and N-methylisosalsoline. These alkaloids are isoquinolines that have not previously been characterised for their molluscicidal activity. The N-methylisosalsoline possesses the highest molluscicidal activity (LC50 = 0.47 ìM against G. truncatula).