食药用菌功能性β-葡聚糖荧光法测定研究

以酵母功能性β-1,3/1,6-葡聚糖为对照品,利用苯胺蓝和β-1,3/1,6-葡聚糖特异结合荧光特性,研究了葡聚糖荧光法测定时的各影响因素,建立了荧光法测定食药用菌功能性β-葡聚糖的方法。pH9.6缓冲液,80℃条件下避光反应15min,室温30min冷却后,398nm激发波长,508nm发射波长,20℃下进行荧光测定。在测定浓度2-20μg/mL范围内,荧光强度与浓度具有良好的线性关系(R2=0.9977),其中检出限为45μg/L,测定精密度和加样回收率良好,相对标准偏差(RSD)分别为1.86%和3.40%,并与酶法进行了比对验证,一致性良好,且荧光法更为节约时间和成本,并对灰树花菌、巴氏蘑菇、香菇和鲍氏针层孔菌四种食药用菌β-葡聚糖提取样品进行了葡聚糖纯度和提取率测定。

Fluorometry method for quantitative determination of functional β-glucans from edible and medicinal mushrooms

Based on the fluorescent property of specific integration of aniline blue dye and standard functional yeast β-1,3/1,6-glucan, some influencing factors of glucan fluorometry were investigated and β-glucan fluorescent determination method was developed. Under the conditions of buffer solution of pH 9.6, aniline blue reaction on yeast β-1,3/1,6-glucan at 80℃ for 15min in dark, then cooling at room temperature for 30min and fluorescent analysis at 20℃ with an excitation maximum wavelength of 398nm and an emission maximum wavelength of 508nm, the standard curve had a good linearity with a concentration range of 2-20μg/mL (R2=0.9977), and the detectable limit was 45μg/L. The precision and recovery of fluorescent method were both measured with a relative standard deviation (RSD) as 1.86% and 3.40%. The method was also validated by enzyme method and showed preferable time saving and economization of cost. The purity and yield of extracted β-glucans samples of Grifola frondosa, Agaricus blazei, Lentinula edodes and Phellinus baumii were analyzed by the method.