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摘要:目的是为了建立一种准确、快速测定姬菇多糖含量的方法,本实验采用蒽酮- 硫酸法测定姬菇多糖含量并对其测定的条件进行优化。通过热水浸提法从姬菇中提取水溶性多糖,并用离子交换柱以及凝胶过滤柱对姬菇多糖进行分离纯化,并测定其分子量。 结果表明:姬菇多糖粗品用Savage法脱除蛋白后,采用DEAE-Cellulose52和Sepharose 4B柱层析纯化,得较为均一的姬菇多糖(PCNPⅠ)。最后根据用不同分子量标准葡聚糖做出的标准曲线,测定出PCNPⅠ分子(Mw)3134873.6±10453.3。本方法准确、快速、重复性好,完全能满足常规分析要求。 关键词:姬菇;多糖;分离纯化;分子量测定
Abstract:Anthrone-sulfuric method was used to determine content of polysaccharide in Pleurotus cornucopiae and the conditions of colourreaction of anthrone-sulfuric method were optimized.The experimental results showed that the RSD in accuracy experiment is1.6%. A water-soluble polysaccharide from Pleurotus cornucopiae (PCNP)was obtained by boiling water extraction and ethanol precipitation,followed by consecutive purification on DEAE--Cellulose52 column and Sepharose 4B column,and was subjected to freeze-drying.The elution profile of PCNP on Sepharose 4B column presented only a single symmetrical peak,indicating that the PCNPⅠwas purified to a homogeneous polysaccharide.The polysaccharide content in dried. PCNPⅠ powder was determined by anthrone-sulfuric acid assay,and no protein components were detected in the powder.The molecular weight of the PCNPⅠ was calculated as 303773.6D,according to the calibration curve of standard dextrans. The method is rapid, accurate, sensitive and suitable forgeneral analysis. Key words: Pleurotus cornucopiae; polysaccharides; isloation and purification; determination of molcular weight
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