| This thesis investigate the isolation and determination of polysaccharides in a Chinese herbs complex prescription used in clinical study for malignant tumor chemotherapy by Chengdu university of traditional Chinese medicine, claimed to have outstanding effect to attenuate adverse effect and enhance curative effect.First, the Chinese herbs complex prescription (CHCP) oral drug was systematically pretested. It was found that there were amino acid, lactone, coumarin, coumarin glycoside, alkaloid, saponin, saccharide, polysaccharide, tannin, phenolic and cardiac glycoside in it. Crude polysaccharides in the CHCP oral drug were isolated preliminarily by multiple deposition in ethanol of 75%~25%. Micro molecule components in crude polysaccharides were removed by dialyze. Four fractions of Polysaccharides (P,P1,P2,P3) qualitatively determined by phenol- sulfuric acid method were eluted out in turn from DEAE- cellulose column chromatography with H2O, 0.5mol/L NaHCO3, 1 mol/L NaHCO3 and 0.5mol/L NaOH. A polysaccharide, HMP-2, was isolated from P1 by Sephadex G-150 column chromatography, with a purity of 97% by HPLC and the structure of ?-furanose uronic acid glycoside identified by IR. P was a mixture of two polysaccharides, which have proximate molecular weight by HPLC and cannot be separated by SephadexG-150 column chromatography.It was also found that first derivative spectrometric value of polysaccharide -phenol- sulfuric acid reaction system is directly proportional to the content of polysaccharides. Basing this, a phenol-sulfuric acid derivative spectrometry for determination of polysaccharides in Chinese herbal medicine is proposed and proved to be an easy, rapid and accurate method with good selectivity, small amount of sample required and little interference of sample matrix color. The method is applied to determination of the content of polysaccharides in the CHCP oral drug (1) and single herbs such as Glycyrrhiza, Astragalus, Angelica etc. with satisfied results.
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