高效液相色谱法测定天然冬虫夏草和蛹虫草及其人工培养物中3个核苷类化合物含量

目的:建立快速分析天然冬虫夏草、蛹虫草及其发酵制品中核苷类化合物成分的高效液相色谱法,比较天然冬虫夏草和蛹虫草及其发酵制品中核苷类化合物的含量。方法:采用超声提取的方法,制备供试品溶液。色谱柱为Hypersil C₁₈柱(200 mm×4.6 mm, 5 μm),流动相为 pH 6.5磷酸盐缓冲液-甲醇(85:15),检测波长为260 nm。结果:天然冬虫夏草和蛹虫草及其人工培养物中的3种核苷类成分在25 min内达到基线分离;腺苷在5.0~80.0 μg·ml^-1范围内与峰面积呈现良好的线性关系,相关系数为0.999 3,样品回收率为99.4%;虫草素在4.37~70.0 μg·ml^-1范围内与峰面积呈现良好的线性关系,相关系数为0.999 7,样品回收率为98.3%;腺嘌呤在6.75~108.0 μg·ml^-1范围内与峰面积呈现良好的线性关系,相关系数为0.999 7, 样品回收率为98.9%。结论:天然虫草与人工虫草培养物有相似的核苷类化合物,但在含量上有较大差异。此方法简便、准确,重复性好,可作为判定天然虫草及其培养物质量的一种方法。

Contents of three nucleosides determined by HPLC in natural and cultured Cordyceps sinensis and Cordyceps militaris

OBJECTIVE To establish a high performance liquid chromatography (HPLC) method for determination of nucleosides from natural and cultured Cordyceps sinensis and Cordyceps militaris, and compare their contents.METHODS The contents of nucleosides were determined using HPLC on Hypersil C₁₈ column with pH 6.0 phosphate-methanol (85:15) as mobile phase and detected at 260 nm.RESULTS Under the optimized conditions, the three nucleosides reached baseline separation in 25 minutes. Adenosine, Cordycepin and Adenine showed good linear relationship in the range of 5.0-80.0 μg·ml^-1 (r=0.999 3), 4.37-70.0 μg·ml^-1 (r=0.999 7) and 6.75-108.0 μg·ml^-1 (r=0.999 7), respectively.CONCLUSION Similar nucleosides have been obtained from natural Cordyceps sinensis and cultured Cordyceps militaris, but with relatively great differences in their contents. This method can be utilized for quantitative analysis of nucleosides in natural and cultured Cordyceps and its substitutes, and it is helpful to control their quality. The method is accurate, fast, and convenient.