QUANTITATIVE DETERMINATION AND PREPARATIVE ISOLATIO N OF TWO MAJOR ALKALOIDS FROM THE VIETNAMESE MEDICINAL HERB EVODIAE FRUCTUS
- Vol.11(11)
- M P Innovare Academic Sciences Pvt Ltd 2019
- 26-32p.
Objective : To develop a simple and accurate HPLC-DAD method fo r 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 C 18 column (150 mm × 4.6 mm I.D., 5 μm), detected at 210 nm. The mobile phase consisted of a cetonitrile (A), methanol (B), and water (C). The v alidated 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 the se compounds were identified using 1 H NMR and 13 C NMR. There was a significant correlation between alkaloid content and fruit size, with a Spe arman correlation coefficient of>0.5 (p<0.001), and there was a large difference in alkaloid contents between three maturity degrees of the frui t. Open-mouth fruits and fruits with average sizes of 4 to 6 mm had the highest alkaloid contents, whereas closed-mouth fruits had the lowes t. Conclusion: This study provided information on the standardizat ion and quality control of evodiamine and rutaecarp ine in Evodiae fructus , as well as a foundation for further pharmacological and tox icological studies.