HPLC波长切换技术同时测定息喘丸中补骨脂素、佛手柑内酯、去甲蟛蜞菊内酯和蟛蜞菊内酯的含量

Simultaneous Determination of Psoralen,Bergapten,Desmethylwedelolactone and Wedelolactone in Xichuan Pills by HPLC Wavelength Switching Technology

目的：建立HPLC法同时测定息喘丸中补骨脂素、佛手柑内酯、去甲蟛蜞菊内酯、蟛蜞菊内酯4个有效成分的含量。方法：采用依利特C18色谱柱（250 mm ＆#215;4.6 mm,5μm）；以甲醇-乙腈（2∶1）与0.5%冰醋酸溶液为流动相进行梯度洗脱；流速为1.1 ml·min-1；柱温为25℃；检测波长（0-20 min、在222 nm波长下检测补骨脂素和佛手柑内酯；20-40 min、在351 nm波长下检测去甲蟛蜞菊内酯和蟛蜞菊内酯）。结果：4个有效成分的线性范围分别为：补骨脂素5.380-107.600μg·ml-1（ r=0.9995）、佛手柑内酯6.870-137.400μg·ml-1（r=0.9997）、去甲蟛蜞菊内酯4.580-91.600μg·ml-1（r=0.9992）、蟛蜞菊内酯7.300-146.000μg·ml-1（r=0.9999）；平均回收率分别为96.82%、98.81%、99.06%、98.39%,RSD分别为0.77%、1.40%、1.20%、0.80%（n=6）。结论：该方法操作准确稳定、灵敏度高、重复性好,可用于息喘丸中上述4个有效成分的定量测定。

Objective：To develop an HPLC method for the simultaneous determination of four active ingredients （ psoralen, ber-gapten, desmethylwedelolactone and wedelolactone） in Xichuan pills. Methods： A Hypersil C18 column was used for the chromato-graphic separation of the four analytes. The mobile phase consisting of methanol-acetonitrile （2∶ 1） and 0. 5% glacial acetic acid solu-tion was used for gradient elution. The flow rate was 1. 1 ml·min-1 . The column temperature was 25℃. Psoralen and bergapten were detected at 222 nm, and desmethylwedelolactone and wedelolactone were detected at 351 nm. Results：There was good linear relation-ship between the concentration of psoralen, bergapten, desmethylwedelolactone and wedelolactone and peak area within the concentra-tion range of 5.380-107.600 μg·ml-1（r =0.999 5）,6.870-137.400 μg·ml-1（r =0.999 7）,4.580-91.600 μg·ml-1（r =0. 999 2） and 7. 300-146. 000 μg·ml-1（r=0. 999 9）,respectively. The average recovery（RSD） was 96. 82%（0. 77%）, 98. 81%（1. 40%）,99. 06%（1. 20%） and 98. 39% （0. 80%）, respectively （n=6）. Conclusion：The developed method is accurate, highly sensitive and well reproducible, which can be used for the determination of the four active ingredients in Xichuan pills.