The Studies On The Extraction, Purification And Determination Of Active Compositions In Solanum Nigrum L.

The extraction, purification and analysis methods of the biologically active composition polysaccharides and TSA in S.N.L. were discussed.(1) Technological microwave-assited extraction process of polysaccharides and TSA was experimented, which excel the traditional reflux extraction in many respects. The solvent of distilled water was used to extract from S.N.L under 325W for a duration of 15 min;ratio of feed-stock weight to the volume of solvent was 1:20; the yield of polysaccharides was 1.94%; the extraction mechanism was also investigated. And the yield of TSA was 26.38μg/g when extracted with 1:20 of 95% ethanol aqueous, at 455W for 8 min. The effect of microwave radiation on the structure of TSA was discussed by TLC and the extraction mechanism was investigated as well.(2) Analysis methods of polysaccharides, protein and clarity were discussed as well as that of TSA. The content of polysaccharides was determined by phenol-sulfuric acid method combined with DNS method and the analysis condition was as follows: for the saccharides, the linear determination range is 18.0⁶6.0μg/mL; for the monosaccharides, the linear determination range is 0.40².00mg/mL. The modified CBB method for determinating the content of protein was studied and the calibration curve was A=-0.0604+0.84543x (R=0.9997). The clarity of extraction solution was analysed by determinating T₆60/% and A₄40 before and after the purification treatment. And the acidic dye colorimetry for determinating the content of TSA was also studied: the linear determination range is 3.20¹6.0μg/mL.(3) Technological process of purification for polysaccharides and elementary characterization on the finally obtained preparation were studied.The unsolvable impurity was removed by high-speed centrifugation at 4000r/min for 10min.Absorption and clarification were completed by the natural ZTC1+1— II clarifier combined with NKA-9 resin and the optimum condition was as follows: the adding sequence is first B and then A; the dosage is 6%(v/v) B and 3% A; the reacting temperature is 80 °C for B while 60°C for A; the concentration of solution was 1:5(g/mL); the elution volume was 3.2mL for every lmL specimen; the dosage of dry resin was 0.4 g for every lmL specimen. And the result shows T?56o rises from 5.40% to 88.8% while the A440 declines from 2.69 to 0.258.Polysaccharides was then deproteinized by Trypsin hydrolyzation combined with Sevag method and the following was the optimum conditions: the specimen was hydrolyzed at 46°C and 6.0pH for 7.5h after adding 2%(v/v) trypsin before extracting for 7 times in Sevag method. The result shows the content of residual protein is only 0.0940% and the clarifier was high to 99.89%.Technological process of further isolation and purification was completed by chromatographic column. Employed with 16X500 mm column, the installation was simple and the adsorption and elution was easy to operate. In this experiment, four fine fractions SNL1, SNL2, SNL3 and SNL4 were obtained by gradient elution with the PB of 7.6pH and NaCl solution through the DE-52 cellulose column chromatography before dialysis and ethanol precipitation. Their homogeneity was further examined by the Sephadex G-100 column chromatography.Physical&chemical characters and homogeneity of the four fine preparations were analysed as well as the structures was investigated by UV and FT-IR spectra.