蛹虫草多糖单糖组成的方法学研究

建立1-苯基-3-甲基-5-吡唑啉酮（PMP）柱前衍生HPLC法测定虫草多糖中单糖组分的通用性方法。本试验以9种单糖标准品为研究对象,通过单因素和响应面试验对衍生反应的温度、时间、pH和PMP用量进行了优化,并运用改进的方法测定了3批不同产地的蛹虫草多糖的单糖组分。结果表明：PMP与单糖摩尔比12:1、温度80℃、时间60min、 pH 8.2为最佳衍生条件,方法学验证证明该方法稳定、可靠、重现性良好。3批蛹虫草多糖中均稳定检测到甘露糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖,摩尔比约为3.8:1.6:5.4:5.8:1,研究结果可为改善虫草多糖成分分析和构效关系研究提供支持。

Methodological study on the monosaccharide composition of polysaccharides in Cordyceps militaris

A universal method for determination of monosaccharide components of Cordyceps polysaccharide was established by using PMP pre-column derivatization HPLC method. Using nine kinds of monosaccharide standards as the object, the derivative reactive conditions including temperature, duration, pH and PMP derivatization were optimized through single factor and response surface test. The optimized method was then used for determining the monosaccharide composition of polysaccharides in Cordyceps militaris from three different regions. The optimal derivatization conditions were summed up as follows: the molar ratio of PMP to monosaccharide was 12:1; the optimal temperature was 80°C; the reactive duration was 60min, and the optimal derivatization pH was 8.3. The methodology validation has proved that this method is stable, reliable and repeatable. Mannose, galacturonic acid, glucose, galactose and arabinose were detected stably in three batches of Cordyceps militaris polysaccharides, and their molar ratio was 3.8:1.6:5.4:5.8:1. The result provides technical support for monosaccharide composition analysis and evaluation of structure-activity relationship of Cordyceps polysaccharide．