Study On The Extraction Isolation And Determination Of Bamboo-leaves Polysaccharide

Bamboo is perennial evergreen plant, belongs to Bambuaoideae Nees of the family poaceae. It has been used for food and medicines for a long time in China. Now the study on bamboo are focused on flavor, the importance of polysaccharide are overlooked. The study take Phyllostachys pubescens as materials, studied bamboo polysaccharide's extraction purification and the activity of antitumor. The effect on extraction efficiency were investigated by means of single factor and orthogonal experiments and Compared the effects of water extraction. Ultrasonic wave extraction, microwave extraction on extraction efficiency. Some powdery BPS were gained by using Sephadex and DEAE column chromatography and comfirmed its composition by using HPTLC and GC and the activity of antitumor is studied by using High-throughput Screening technology. The results as follows:1 The method of phenol-sulfuric acid was developed. The results showed there was a good linear relationship with R~2=0.9989 between the absorbance and the concentration within the range from 20 μg/ml to 100μg/m when the wavelength was 490 nm.2 The parameter of extract technology was perfected, and the technology of ultrasonic and microwave was applied to the bamboo polysaccharide's extraction for the first time. The optimal water extraction parameters were as follows: temperature 80℃, extracting 3 times, each time 90min, solid-liquid ratio 1 ∶ 25; The optimal ultrasonic extraction parameters were as follows temperature 70℃, extracting 3 times, each time 20min, solid-liquid ratio 1 ∶ 20,The optimal microwave extraction parameters were as follows: power 500W, extracting 3 times, each time 2min, solid-liquid ratio 1∶15. The yield of bamboo polysaccharide was 0.661%,0.708% and 0.655% respectively.3 The crude polysaccharides were deproteinizated by Sevag method. Then they were purified on DEAE-cellulose, followed by an elution with water, aqueous sodium chloride(0.05M, 0.1M, 0.3M, 0.5M) and Fractions were examined with phenol-sulfuric acid method. The fractions were divided into four groups, named as BPS1,BPS2, BPS3, BPS4 and the yield is 46.45%, 4.31%, 1.62%, 0.53% separately. The BPS1 were further purified by a column chromatogram of DEAE-cellulose, Sephadex G-75 to get the BPS 111. Their homogeneity were proved by Sephadex G-100 chromatography. According to HPTLC and GC the BPS111 was composed of rhamnose: arabinose: xylose: mannose: glucose: galatose=2.91: 9.27: 4.42: 6.38: 25.59: 9.44.4 Bamboo polysaccharide's activity of antitumor is studied by using High-throughput Screening technology, the results showed when Bamboo polysaccharide's concentration is 10 μ g/ml have function on THP-1.