UV-VIS与HPLC对牛黄类药材中胆红素含量的测定方法及影响因素分析

Detection Method and Influence Factors of Bilirubin in Bovis Calculus by UV-VIS and HPLC

目的:选择和建立对牛黄类药材中总胆红素含量测定的适宜方法。方法:采用HPLC法对牛黄、体外培育牛黄中胆红素含量进行测定,HPLC法条件为Boston Green ODS-C18(150 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-1%冰乙酸溶液(95∶5),流速为1.0 ml/min,检测波长450 nm,柱温30℃;并与UV-VIS法比较其优劣。结果:牛黄类药材中总胆红素含量结果显示,UV-VIS法较HPLC法高,牛黄比体外培育牛黄的差异大;超声温度对不同类型牛黄中胆红素的提取效率影响不同。结论:HPLC法比UV-VIS法更具适用性与专属性,可准确得到牛黄类药材中胆红素含量,且操作简便、快速,应用性较广。

Objective：To compare the UV-VIS method and HPLC method applied in determining bilirubin content of Boris Calculus and to select a appropriate method. Factors affecting the accuracy are also optimized. Methods： UV-VIS method and HPLC method were applied to study the Bovis Calculus and Bovis calculus sativus in bilirubin content. The separation was performed on a Boston Green ODS-C18 （ 150mm ×4.6mm, 5μm） column, the mobile phase was acetonitrile-l% glacial acetic acid （95 ： 5 ） at a flow rate of 1.0ml/min, the UV detection was set at 450nm and the column temperature was 30℃. Results ： The values that UV method collected was higher than that of the HPLC method in bilirubin content of Boris Calculus, and the difference in Bovis Calculus was more obvious than that in Bovis calculus sativus; the temperature of ultrasound has different effect on bilirubin extracting of different kinds of bezoars. Conclusion： HPLC method is more suitable and specific than UV method ; it can accurately determine bilirubin content of bezoar and is simple, rapid, widely applied.