HPLC-RID测定冬虫夏草及发酵虫草制剂中虫草酸的含量

目的：建立发酵虫草制剂中虫草酸的高效液相色谱含量测定方法。方法：采用正向氨基色谱柱Agilent Zorbax NH₂（250 mm×4.6 mm，5 μm），以乙腈-水（78：22）为流动相，流速1.0 mL·min^-1，柱温30 ℃，示差折光检测器检测（内部温度35 ℃）。结果：虫草酸进样量在20.19~100.95 μg范围内与峰面积呈良好的线性关系，平均加样回收率（n=6）为98.5%，RSD为0.85%。金水宝胶囊中虫草酸含量为2.29%~3.77%，均值为2.86%；百令胶囊中虫草酸含量为2.42%~5.28%，均值为3.68%；心肝宝胶囊中虫草酸含量为3.78%~5.02%，均值为4.37%；至灵胶囊中虫草酸含量为3.63%~5.12%，均值为4.17%；宁心宝胶囊中虫草酸含量为2.37%~4.08%，均值为3.37%，不同企业的制剂中虫草酸含量差异较大；虫草头孢菌粉系宁心宝胶囊的原料药，此次测定的样品中虫草酸的含量为2.30%~3.27%。可见，这种多厂家生产原料药，多厂家生产制剂的现状导致宁心宝胶囊工艺、原料难统一，标准亟待完善和提高。结论：经方法学验证，本方法可用于测定冬虫夏草及发酵虫草制剂中虫草酸的含量，亦可用于控制药材及制剂的质量。

Objective: To establish a method for determination of cordycepic acid in Cordyceps sinensis and fermented Cordyceps preparation.Methods: HPLC-RID was conducted on a Agilent Zorbax NH₂ column(250 mm×4.6 mm,5 μm)with a mobile phase of acetonitrile-water(78∶22)at the flow rate of 1.0 mL·min^-1.The column temperature was maintained at 30 ℃,and refractive index detector was used to detect cordycepic acid with the internal temperature was 35 ℃.Results: The linear response was 20.19-100.95 μg for cordycepic acid.The average recoveries (n=6)for cordycepic acid was 98.5% and the RSD was 0.85%.The contents of cordycepic acid were 2.29%-3.77% in Jinshuibao capsules with the average content of 2.86%,2.42%-5.28% in Bailing capsules with the average content of 3.68%,3.78%-5.02% in Xinganbao capsules with the average content of 4.37%.The contents of cordycepic acid were 3.63%-5.12% in Zhiling capsules (the average content was 4.17%),2.37%-4.08% in Ningxinbao capsules,and 2.30%-3.27% in fermented powder(Cephalosporium powder),respectively.The contents of cordycepic acid were different due to the different production process and different sources of fermented powder for fermented Cordyceps preparation of many manufacturers.It is necessary to improve and strengthen the quality control of fermented powder,so as to set reasonable cordycepic acids limits.Conclusion: By methodology verification,the established method can be applied for the determination of the content of cordycepic acid from fermented Cordyceps preparation,which will be benefit to the quality control of the drugs.