酵母β-葡聚糖是一种应用价值较高的多糖,因其不溶于酸碱,在测定其含量时,存在着一定的困难。刚果红可与酵母β-葡聚糖发生特异性结合,用于定量检测分析。实验对刚果红检测酵母β-葡聚糖的反应条件及反应体系进行优化,结果显示:在0.1 mol/L pH 7.5的磷酸盐缓冲液中,0.2 mg的刚果红溶液可与4~1 600 μg内的β-葡聚糖发生反应,产生的复合物较刚果红发生红移,在540 nm下有最大吸收差值;最佳反应条件为20 ℃,反应15 min;酵母β-葡聚糖含量在4~200 μg时,与吸光度之间有良好的线性关系,其线性回归方程为:Y=0.001 15X-0.001 14,R2=0.998 6;精密度试验相对标准偏差为1.154%;平均回收率为100.32%,并不受碱溶性β-1,3葡聚糖的干扰,具有较高的灵敏度,可用于酵母β-葡聚糖的检测。
The yeast β-glucan is a valuable polysaccharide with a wide range of applications, and because it is insoluble in acids and alkali, there are difficulties in determining its content. Congo red can specifically bind to yeast β-glucan and can be used for quantitative assay analysis. The experiment was carried out to optimize the reaction conditions and reaction system for the detection of yeast β-glucan by Congo red, and the results showed that 0.2 mg of Congo red solution in 0.1 mol/L pH 7.5 phosphate buffer could react with the content of yeast β-glucan in the range of 4-1 600 μg. The UV-Vis spectroscopy results indicated that the absorption edge of Congo red was redshifted to 540 nm. The optimal reaction conditions were 20 ℃ and 15 min. There was a good linear relationship between concentration in the range of 4-200 μg and the absorbance, and the linear regression equation was Y=0.001 15X-0.001 14, R2=0.998 6. The RSD of the precision test was 1.154%. The average recovery rate was 100.32%, and was not interfered by the alkali-soluble β-1,3 glucan, having high sensitivity, which can be used for the determination of yeast β-glucan.
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