苯酚硫酸法测定萌发菌HL-003多糖方法的研究
Study on phenol-sulfuric acid method for determination of polysaccharides from germination fungus HL-003
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摘要: 为确定苯酚硫酸法测定萌发菌HL-003多糖的最佳参数。选取苯酚浓度和浓硫酸量为自变量,检测不同苯酚浓度和浓硫酸量下的最大吸收波长和吸光值,从而确定最佳的苯酚浓度和浓硫酸量,在此基础上检验方法的显色时间、精密度、线性范围和回收率。结果表明:最佳苯酚浓度为5%,浓硫酸量为3.5 m L,最大吸收波长为488 nm,显色时间15 min;葡萄糖浓度在0200 mg/m L范围内线性良好;在优化方法下进行精密度实验,葡萄糖、胞内多糖和发酵液多糖测定结果的相对标准偏差（RSD）分别为0.474%、1.015%和0.712%,测定结果稳定可靠;本方法对葡萄糖和葡聚糖的回收率均在99.10%以上,回收率高,与真实值吻合;通过测定,萌发菌HL-003胞内和发酵液多糖的含量分别为2.028%和2.998 g/L。Abstract: In order to determine polysaccharides of germination fungus HL- 003, parameters of the phenol-sulfuric acid method were optimized. Phenol concentration and volume of concentrated sulfuric acid was the independent variable, the maximum absorption wavelength and absorbance were detected under different concentrations of phenol and concentrated sulfuric acid volume. Immediately,the color of time,precision,linearity and recovery were measured in order to determine the optimum parameters. The results showed that phenol concentration of 5%,concentrated sulfuric acid content of 3.5 m L,the maximum absorption wavelength of 488 nm,coloration time 15 min. The methods had a high precision and recovery results were reliable,and consistent with the true value. The concentration of glucose in the range of 0~200 mg/m L had a good linearity.Intracellular and fermentation of polysaccharide contents were 2.028% and 2.998 g/L,by detecting with optimized method.
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