不同种类牛黄投料的小儿化毒胶囊(散)中胆红素的研究

Study on Bilirubin in Xiaoer Huadu Capsule Made of Different Calculus Bovis

目的:建立"小儿化毒胶囊(散)"中游离胆红素、总胆红素以及部分胆红素的HPLC测定方法,比较以人工牛黄、体外培育牛黄2种牛黄分别投料的小儿化毒散之间差异。方法:分别以二氯甲烷、10%草酸溶液(含0. 15%十六烷基三甲基氯化铵)-二氯甲烷、0. 2 mol/L乙二胺四醋酸二钠-二氯甲烷提取样品中游离胆红素、部分胆红素、总胆红素,用HPLC法测定,固定相为C_(18)色谱柱,流动相为乙腈-1%冰乙酸(95∶5),检测波长450 nm,柱温为25℃,流速为1. 0 m L/min。结果:胆红素在5. 64～282 ng范围内线性关系良好,游离胆红素、部分胆红素、总胆红素平均加样回收率为90%～95%,RSD分别为4. 2%、4. 1%和4. 9%,2种牛黄投料的小儿化毒散中胆红素含量及形态存在明显差异。结论:建立的方法可有效识别"小儿化毒胶囊(散)"中牛黄的投料种类,为其他含有牛黄的中药制剂质量评价提供了借鉴。

Objective: To establish an HPLC methods for free,portion and total bilirubin in Xiaoer Huadu Capsule(Powder)and compare the difference between 2 types of Calculus Bovis. Methods: The free,portion and total bilirubin were extracted with dichloromethane,10% oxalic acid solution(containing 0.15% cetyltrimethylammonium chloride)-dichloromethane,0.2 mol/L disodium ethylenediaminetetraacetate-dichloromethane,determined by HPLC with C 18 column,the mobile phase was acetonitrile-1%glacial acetic acid(95∶ 5).The detection wavelength is 450 nm,and the column temperature was 25 ℃.The flow rate was 1.0 mL/min. Results: Bilirubin showed good linear relationship in the range of 5.64 ng-282 ng,and the average recoveries of free,portion and total bilirubin was 90%-95%,with RSD 4.2%,4.1% and 4.9%.There were significant differences in the content and form of bilirubin of 2 kinds of Calculus Bovis. Conclusion: The method established is easy to operate with high effectiveness in identifying bezoar in Xiaoer Huadu Jiaonang and other Chinese traditional patent medicines containing Calculus Bovis.