反相高效液相色谱法测定藏药材红毛五加中金丝桃苷的含量

Determination of Hyperoside in Acanthopanacis giraldii by RP-HPLC

目的建立藏药材红毛五加药材中金丝桃苷的含量测定方法并比较其不同药用部位及不同产地、不同采收期的红毛五加皮中金丝桃苷的含量。方法Kromasil C18柱(4.6mm×250mm,5μm),柱温30℃,流动相甲醇(A)-0.5%磷酸水溶液(B),梯度洗脱,程序为:基线(A∶B=30∶70),0～5min(A∶B=35∶65),5～15min(A∶B=40∶60),15～30min(A∶B=50∶50);检测波长358nm,流速1ml.min-1。结果金丝桃苷在2.90×10-3～0.58μg范围内峰面积与进样量具有良好的线性关系,回归方程Y=4×106X-14065,r=0.9999(n=9),平均回收率为100.6%,RSD为0.96%(n=6)。结论该法简便、准确、重复性好,有助于红毛五加质量控制方法的研究。

Objective To develop an RP - HPLC method for determination of Hyperoside in Acanthopanax giraldii Harms, and to determine the contents of Hyperoside in different parts, different areas and different collection time of Acanthopanax giraldii Harms. Methods The analysis was carried out at 30 ℃ on a Kromasil Cls column eluted with a mobile phase consisted of methanol （A） - 0.5 % phosphoric acid （B）. The gradient elution was as following ： i The ratio of A and B was 35/65 from 0 to 5 min. ii The ratio of A and B was 40/60 from 5 to 15 min. iii The ratio of A and B was 50/50 from 15 to 30 min. The detective wavelength was at 358 nm with the flow rate of 1.0 ml/min. Results The calibration curve was linear within the concentration range of 2.90 × 10 ^- 3 - 0.58 μg with r of 0.999 9 （ n = 9 ）. The average recovery was 100.6% and RSD was 0.96% （ n = 6 ）. Conclusion This method is simple, accurate and reproducible. It is useful for quality control of Acanthopanax giraldii Harms.