HPLC测定人工蛹虫草中6种核苷类成分的含量

Determination of the nucleosides in cultured Cordyceps militarisby HPLC

目的建立人工蛹虫草中核苷类成分的HPLC测定方法,为产品质量标准提供参考。方法用高效液相色谱法测定人工蛹虫草中的有效成分,选用Waters XBridgeTMC18柱色谱柱（250×4.6 nm,5μm）;流动相为甲醇-0.2%磷酸二氢钾溶液梯度洗脱,流速为1.0 m L/min,检测波长为260 nm,柱温为30℃。结果虫草素、腺苷、鸟苷、尿苷、胸苷、胞苷分别在0.025~0.5 mg/m L,0.01~0.2 mg/m L,0.01~0.2 mg/m L,0.01~0.2 mg/m L,0.01~0.2 mg/m L,线性关系良好,平均回收率虫草素为99.34%,胞苷为99.04%,尿苷为99.09%,鸟苷为98.75%,胸苷为102.38%,腺苷为99.27%。结论该测定方法简便易行、准确性好、灵敏度高、重复性好,可用于人工蛹虫草中核苷类成分的质量控制。

Objective To determine the content of nucleoside in cultured Cordyceps militaris. Methods The HPLC method was established for the determination of the content of nucleosides in cultured Cordyceps militaris,The HPLC was performed on Waters XBridgeTMC18（ 250 mm × 4. 6 mm,5 μm）column,mobile phaseconsisted methanol-0. 2% monopotassium phosphate gradient elute. column temperature 30 ℃,and flow rate was 1. 0 m L / min. The detection wavelength was 260 nm. Results A good linearity was obtained in the range of 0. 025-0. 5 mg / m L of cordycepin,0. 01-0. 2 mg / m L of cytidine,0. 01-0. 2 mg / m L of uridine,0. 01-0. 2 mg / m L of vernine,0. 01-0. 2 mg / m L of thymidine and 0. 01-0. 2 mg / m L of adenosine. The average recovery rates of cordycepin,cytidine, uridine, vernine, thymidine,and adenosine were 99. 34%,99. 04%,99. 09%,98. 75%,102. 38% and 99. 27%,respectively. Conclusion The established method is simple,accurate and sensitive,and suitable for quality control of cultured Cordyceps militaris.