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Studies on Isolation, Purification and Chemical Structure for Polysaccharides from Camellia Chrysant

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Tutor: LinHuaJuan;QinXiaoMing
School: Guangdong Ocean University
Course: Of Food Science
Keywords: Camellia chrysantha (Hu) Tuyama,Polysaccharide,Isolation and purification,Chemic
CLC: S567.19
Type: Master's thesis
Year:  2011
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Abstract:
Camellia chrysantha£¨Hu£© Tuyama is a kind of plant which in camellia genus. It is called¡°the queen of the camellia¡¯s world¡±and¡°the ailuropoda of the plant¡¯s domain¡±because it has flavous petal which is the unique variety in the camellia . As a orthodox chinese herbal medicine in Guangxi Zhuang nation Camellia chrysantha£¨Hu£© Tuyama has many physiological function such as depressing fat, sugar and cholesterol in blood serum ,restraining tumor, elevating organism immunity and so on. tea polysaccharide from the leave of Camellia chrysantha £¨Hu£© Tuyama is considered to be one of the important effect components, however its chemical structure is unclear. In this article, the isolation¡¢purification and the chemical structure of tea polysaccharide from the leave of Camellia chrysantha £¨Hu£© Tuyama was studied, this provides an important scientific theory basis for the structure-activity relationships research of tea polysaccharide.The main contents and conclusions were listed as followed:1¡¢Studied of some major components in the concentrated extraction from leaves of Camellia chrysantha £¨Hu£© Tuyama and initial separation of tea polysaccharide from the leave of Camellia chrysantha £¨Hu£© Tuyama. some major components in the concentrated extraction £¨Total sugar, saponins, tea polyphenol and flavonoids£© were assaied, then a crude polysaccharide matter £¨TPS£© was obtained from leaves of Camellia chrysantha £¨Hu£© Tuyama by alcohol precipitation. Sugar analysis showed that the TPS consisted of neutral sugar and acid sugar£¨The contents was 41.9% and 24.2% respectively£©, and the content of neutral sugar was two times of galacturonic acid, implying TPS was some pectins with high ratio of neutral sugar. Sugar composition analysis showed that glucose, galactose, arabinose, mannose and rhamnose were the major compositions in TPS£¨Its glycosyl ratio was 31%¡¢27%¡¢21%,13% and 6%£©, and the results showed that TPS was a complicated composite which contained diversiform polysaccharides.2¡¢With distilled water¡¢0.1 mol/L¡¢0.2 mol/L¡¢0.3 mol/L¡¢0.4 mol/L and 0.5 mol/L NH4HCO3 solution salt elution in turn, TPS was purified by negative ion exchange chromatographic column on DE52-cellulose £¨HCO3- form£©. The components named TPS0¡¢TPS1¡¢TPS2¡¢TPS3¡¢TPS4 and TPS5 were gained. Sugar analysis showed that the TPS0¡¢TPS1 and TPS2 mainly contained neutral sugar, while TPS3¡¢TPS4 and TPS5 Contained higher percentages of acid sugar, implying TPS0¡¢TPS1 and TPS2 belong to neutral polysaccharides, while TPS3¡¢TPS4 and TPS5 belong to pectic polysaccharides. Sugar composition analysis showed the glycosyl composition and ratio of TPS0-TPS5 Were significantly different, the TPS3¡¢TPS4 and TPS5 with high proportion of galacturonic acid, its rhamnose ratio £¨2.46% -4.35%£© higher than the TPS0, TPS1 and TPS2 components which had not detected rhamnose, while the glucose content £¨2.69%, 3.76% and 4.63%£© of TPS3, TPS4 and TPS5 was significantly lower than the neutral sugar components TPS0, TPS1 and TPS2 £¨21.1%, 23.28% and 2.19%£©, Speculated TPS3, TPS4 and TPS5 component is likely to be pectin which had the main chai n Composed by galacturonic acid and rhamnose alternately and side chains with more glucose, and another way there may be a mixture of glucan and pectin which had the main chai n and side chains.TPS3 was further purified by Sephacryl S-200 gel column chromatography, And two components named TPS3-1 and TPS3-2 were gained. Sugar composition analysis showed TPS3-1 and TPS3-2 were all contain galacturonic acid and neutral sugar, and the Neutral sugar content Of TPS3-1 was higher than TPS3-2, and the ratio of neutral sugar and galacturonic acid of TPS3-1 and TPS3-2 was 2.2 and 1.3. Sugar composition analysis showed that rhamnose, arabinose glucose and galactose were the major compositions in TPS3-1 and TPS3-2 £¨Its glycosyl ratio respectively was 6.20: 26.27: 2.64: 18.48 and 5.23: 11.57: 18.39: 10.72£©. The molecular of TPS3-1 and TPS3-2 were 415kDa and 338kDa determined by HPGPC. Speculated that TPS3-1 may be RG pectin with main chain composed by galacturonic acid and rhamnose alternately and side chains with arabinose, galactose and a little glucose. TPS3-1 may be a RG pectin polysaccharide which side chain had More glucose.3¡¢The rusults of IR¡¢1H NMR and 13C NMR showed that TPS3-1 and TPS3-2 were pyranose which had¦Á-glycosidic linkage¡¢¦Â-glycosidic linkage¡¢-COOH and methoxyl. And the rusults of methylation and partial hydrolysis with acid analysis showed that the structure of PS3-1 and TPS3-2 were a kind of pectin which similar to rhamngalacturonan-I.
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