Abstract

High-speed counter-current chromatography was used to separate and purify ecdysteroids for the first time from the stems of Diploclisia glaucescens using a two-phase solvent system composed of ethyl acetate–n-butanol–ethanol–water (3:0.2:0.8:3, v/v). Three ecdysteroids were obtained from 260 mg of ethyl acetate extract of the residue obtained after evaporation of the crude ethanolicextractof D. glaucescens in one-step separation, which were identified as paristerone (I, 30.5 mg), ecdysterone (II, 7.2 mg), and capitasterone (III, 8.1 mg) by electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR). Their anti-inflammatory activities were evaluated by measuring the inhibitory ratios of β-glucuronidase release in rat polymorphonuclear leukocytes (PMNs) induced by platelet-activating factor. Compounds I–III showed significant anti-inflammatory activities with IC50-values ranging from 1.51 to 11.68 μM, respectively.

Highlights

  • Diploclisia glaucescens (Menispermaceae) is a medicinal plant widely distributed in Guangxi and Yunnan province, China

  • Phytochemical investigations revealed that ecdysteroidsare the major components of D. glaucescens [2,3,4]

  • 98.3% purity, and 8.1 demonstrate mg of capitasterone with purity determined by HPLC analysis. These results demonstrate the high resolving power of High-speed counter-current chromatography (HSCCC)

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Summary

Introduction

Diploclisia glaucescens (Menispermaceae) is a medicinal plant widely distributed in Guangxi and Yunnan province, China. The leaves and stems of the plant have been used as a folk medicine in the treatment of rheumatism, snake venom, biliousness, and venereal diseases [1]. Ecdysteroids exhibit interesting pharmacological effects on mammals, such as the stimulation of protein synthesis, antidiabetic activity, and antirheumatic activity [5,6,7]. Ecdysteroids are steroid hormones, that play a highly complex ecological role in the chemical communication among the kingdoms [8]. In view of these beneficial biological properties and broad applications, the study on the separation and purification of ecdysteroids from D. glaucescens is necessary

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