酸酶水解-HPLC法检测香菇多糖中β-D-葡聚糖含量

摘要

A new method for determination of β-D-glucans content in Lentinan was developed by acidic and enzy- matic hydrolysis coupled with high performance liquid chromatography （HPLC）. Lentinan was hydrolyzed entirely to glucose with exo-1,3-β-glucanase and β-Glucosidase after partial acid hydrolysis for determination of total glucans by HPLC. α-Glucans in Lentinan samples were hydrolyzed to glucose with Amyloglucosidase and then also analyzed by HPLC. The content of β-D-glucan was calculated by the difference between total glucans and α-glucans. Moreover, the above method and phenol sulfuric acid method and Congo red method were used respectively to determine the con- tent of glucans （or β-D-glucan） in laminarin, curdlan and yeast glucan samples whose β-D-glucan content were known. Compared the results from three methods, acidic and enzymatic hydrolysis coupled HPLC method were the most suitable for quantitative analysis of β-D-glueans in Lentinan. RSD of the method were 2.02% -2.94%.%为了更有效地监控食品和医药行业中香菇多糖制品的质量，建立了一种香菇多糖中主要活性成分β-D-葡聚糖含量测定的新方法：香菇多糖样品经酸部分水解后用β-1，3-葡聚糖外切酶和β-葡糖苷酶进一步完全水解以测定总葡聚糖含量；另用淀粉葡萄糖苷酶水解样品中仪一葡聚糖；根据HPLC测定的总葡聚糖与α-葡聚糖含量之差计得β-D-葡聚糖含量。分别用该方法和苯酚硫酸法及刚果红法测定已知含量的β-D-葡聚糖对照样品的结果表明：该方法最接近于对照品标示值，且重复性好，可用作为香菇多糖产品中β-D-葡聚糖含量测定的专属方法。