反相高效液相色谱法测定康肾颗粒中的葛根素

建立了反相高效液相色谱分离、质谱定性、紫外检测定量测定康肾颗粒中葛根素含量的方法。采用YMC-ODS色谱柱(4.6 mm i.d.×100 mm,10 μm),选用甲醇(含0.1%三氟醋酸)和水(含0.1%三氟醋酸)体系为流动相,非线性梯度洗脱分离,流速1.0 mL/min,检测波长250 nm；用基质辅助激光解吸离子化飞行时间质谱(MALDI-TOF MS)分析鉴定对照品和样品的馏分,在确定目标峰后进行紫外检测定量测定。结果表明，葛根素在0.132 ~1.32 μg范围内其峰面积与进样量呈良好的线性关系（r=0.9997）,平均加标回收率高于103%,相对标准偏差低于2.0%(n=6)。应用该方法测定康肾颗粒中葛根素，其平均含量为3.09 mg/g。该方法简单、快速、重现性好。

Determination of Puerarin in Kangshen Granule by Reversed-Phase High Performance Liquid Chromatography

Kangshen granule, containing 12 kinds of traditional Chinese medicine, has complex composition. A new method for the determination of puerarin in Kangshen granule was developed using reversed-phase high performance liquid chromatography coupled with matrix assisted laser desorption ionization time of flight mass spectrometry and ultraviolet detection (RP-HPLC/MALDI-TOF MS/UV). The sample was separated by RP-HPLC with a non-linear gradient elution, and then was qualitatively detected by mass spectrometry to identify the puerarin peak. The chromatographic process was performed on a YMC-ODS column under the optimized conditions with a mobile phase consisting of methanol, water and 0.1% TFA, and detected at 250 nm. The linear range of puerarin was from 0. 132 to 1.32 microg (r = 0.9997). The recovery was higher than 103% and the relative standard deviation was lower than 2.0% (n=6). The puerarin content in Kangshen granule sample was determined and the puerarin content was 3.09 mg/g. The method is simple and feasible for the determination of puerarin with a good repeatability. The method can be used for quality control of Kangshen granule.