HPLC双波长切换同时测定玄参药材中哈巴苷和哈巴俄苷的含量

Simultaneous detection of the contents of harpagide and harpagoside in Xuanshen by Dual-wave switching HPLC

目的:建立以双波长切换方式同时测定玄参药材中哈巴苷和哈巴俄苷的含量的方法。方法:采用Shimpack VP-ODS C_(18)柱(250 mm×4.6,5μm)色谱柱,以乙腈(A)-磷酸(0.03%)(B)为流动相进行梯度洗脱:0-10 min,3%-10%(A);10-20 min,10%-33%(A);20-25 min,33%-50%(A);25-30 min,50%-80%(A);30-35 min,80%(A);35-37min,80%-3%(A);检测波长:前10 min为278 nm,10-15 min为210 nm,15 min后为278 nm,柱温为30℃,进样量为10μL,流速为1 mL·min^(-1)。结果:哈巴苷在0.011824-0.11824 mg·mL^(-1)范围内与哈巴俄苷在0.00416-0.0416 mg﹒mL^(-1)范围内线性关系良好。结论:该检测方法稳定可行,可用于同时测定玄参药材中哈巴苷和哈巴俄苷的含量。

Objective： To establish Dual-wave switching HPLC method to detect the contents of harpagide and harpagoside in Xuanshen. Method： The analysis was carried on a Shim-packVP-ODS C18 （250 mm×4.6,5μm） column. The mobile was acetonitrile （A） and 0.3% Phosphoric Acid （B） which was in gradient elution at flow rate of lmL.min-1 as following： 0-10min, 3%-10% （A）, 10-20 rain, 10%-33% （A）; 20-25 min, 33%50% （A）; 25-30 min, 50%-80% （A）; 30-35 min, 80% （A）; 35-37min, 80%-83% （A）. The wave length was set at 278 nm during 0-10 min; 210 nm during 10 -15 min; 278 nm after 15 min. Column temperature was 30℃, and sample volume was 10 gL. Result： The response of harpagide in the range of 0.011824-0.11824 mg﹒mL^-1 and the response of harpagoside in the range of 0.00416-0.0416 mg﹒mL^-1 had a good linear relationship with concentration. Conclusion： The detection method is stable and feasible, which can be used for simultaneous determination the contents ofharpagide and harpagoside in Xuanshen.