人工蛹虫草核苷类成分超声提取工艺优化HPLC定量分析

摘要

This paper aims to optimize the ultrasonic extraction conditions and to develop a method by HPLC for simultaneously determining four nucleotides (uridine,guanosine,adenosine and cordyce-pin) in Cordyceps militaris. Optimization of the ultrasonic extraction conditions was conducted by orthogonal layout. The HPLC analysis working conditions are as follows; column was Amethyst C18-H (4.6 mm x250 mm,5μm) ; volume ratio of methanol-water was 15:85,retention time was 18 min; flowing rate was 0. 8mL·min-1; temperature is 35℃; detective wavelength was 260 nm; injection volume was 10μL. The optimum extraction conditions were confirmed as follows; the ratio of sample to water(m/V) was 1 = 30; solvent is pure water; extraction temperature was 25℃; extraction time was 45 min. The HPLC method used in this experiment was simple and convenient,hence it is valuable for quality evaluation of Cordyceps militaris.%利用正交设计优化人工蛹虫草(Cordyceps militaris)中4种主要核苷成分(尿苷、鸟苷、腺苷和虫草素)的超声提取条件,并建立高效液相色谱定量分析方法.高效液相色谱法定量分析采用的色谱条件:Amethyst C18 -H柱(4.6 mm×250 mm,5μm),V(甲醇):V(水)=15:85,停止时间18 min,流速0.8 mL· min-1,柱温35℃,检测波长260 nm,进样量10μL.优化超声提取条件为料液比(m/V)1:30,提取溶剂为纯水,25℃下超声45 min.本试验采用的高效液相色谱定量分析方法简便可靠,重复性、精密度及回收率良好,可以为人工蛹虫草质量控制和综合评价提供参考.