Study On The Sedative And Hypnosis Active Component From Zizyphus Vulgaris Var.Spinosus

In this paper the application of macropore adsorbed resin for extraction and purification of total alkaloids from Zizyphus vulgaris var. spinosus has been studied as well as evaluated the sedative activity of total alkaloids. The bioassay-directed fractionation afforded sedative constituent I and non-active X , which isolated by a solvent method. Structural determination of cyclopeptide alkaloids I is based on UV,IR,MS,~13C-NMR,amino acid auto-analyses and some chemical reactions.Three different extraction methods of total alkaloids have been investigated in our experiment. The acid extraction method has been selected as a final strategy. The polarity and surface area of macropore adsorbed resin were compared to determine the extraction and refinement. Non-polarity X-5 and mid- polarity AB-8 macropore adsorbed resin were chose as well as studied the solid quantity from different concentration alcohol, UV absorption spectrum was detected to establish 70% alcohol was the best wash solvent , it' sproductive rate was 1. 25% ?Constituent I and X was separated by solvent method, and pharmacological experiment to confirm I was sedative and hypnosis active component, it' s productive rate was 0. 073%.The structure of alkloid constituent I was deduced from Dragendorff s reaction and amino acid auto-analyses of acid hydrolysis , it is a cyclopeptide alkloid consist of leucines isoleucine^ProlineoUV spectra of constituent I has two typical spectral bands at 202nm & 270nm0 Infrared spectrum of acive constituent I showed absorptions of Y NH 3280cm"1 , a , |3 -unsatd OO at 1660 cm"1, and absorptions due to phenol ether at 1240 cm"1. 13C~NMR spectrum showed 27 carbon peaks , the appearance of three OO peaks at 6 171. 786, 170. 822 & 167. 023, eight C=C ion peaks at 6 113. 814~157. 476, the other 16 carbons peaks at 683.721 ?2. 1680 The molecular formular of I was determined by molecular ion peak 484 in EIMS to be C^HioNA. Due to the typical fragments for 3-hydroxyproline m/e96 fO 68 , the structure of contituent I belong to amphibine-B with 14-membered cyclopeptide alkloid, which the bonding amino acid was isoleucine, final amino acid was leucine. It is the first time to find this compound in zizyphus vulgaris var. spinosus.