天然牛黄和体外培育牛黄中不同胆红素含量及比例的初步探究  

Preliminary Study on Contents and Ratios of Different Types of Bilirubin in Natural and in Vitro Cultured Calculus Bovis

作　　者：夏晶[1] 曹帅[1] 沙祎炜 杨新华[1] 季申[1] 

机构地区：[1]上海市食品药品检验所,上海201203 [2]上海市崇明食品药品检验所,上海202150

出　　处：《中国药学杂志》2017年第11期971-977,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81473347);上海市科学技术委员会项目(15DZ0502700);国家药典委员会人工牛黄质量标准专项研究(2016-61)

摘　　要：目的建立牛黄中共价胆红素、结合胆红素和游离胆红素的专属分析方法,深入比较各胆红素在天然牛黄和体外培育牛黄中的含量和比例,为后续含牛黄类制剂中牛黄鉴定提供依据。方法样品采用二氯甲烷作为基本提取溶剂,辅助加入不同螯合剂来进行各类胆红素的提取,并采用间接计算法得到牛黄中3种胆红素的量。具体采用HPLC法,迪马Diamonsil C_(18)(4.6 mm×150 mm,5μm)色谱柱;流动相为乙腈-1%冰醋酸溶液(95∶5);检测波长为450 nm。结果游离胆红素在天然牛黄中均有检出但含量较低、在体外培育牛黄中除个别样品外均未检出;天然牛黄中总胆红素含量差异较大,而体外培育牛黄中总胆红素含量较为接近;从共价胆红素和结合胆红素占总胆红素的比例来看,天然牛黄中共价胆红素比例一般较高,体外培育牛黄则以结合胆红素为主。结论天然牛黄和体外培育牛黄中不同类型胆红素的含量及比例不同,具较强专属性,为识别含牛黄制剂中牛黄种类及如何评价牛黄质量提供了新的思路。OBJECTIVE To develop a method for determination of different types of bilirubin in bezoar and explore the differ- enees in their contents or ratios between natural and in vitro cultured Calculus Bovis to provide reference for identification of Chinese patent drugs containing bezoar. METHODS Free bilirubin was determined directly by HPLC, while unconjugated and total bilirubin in bezoar were converted into free bilirubin respectively using different chelating agents. Converted bilirubin was eluted on a Diamonsil Cts column（4. 6 mm×150mm,5μm）using isocratic mobile phase composed of acetonitrile-l% acetic acid （95： 5） and detected at 450 nm. RESULTS A small amount of free bilirubin was detected in Calculus Bovis but not found in in-vitro cultured Calculus Bo- vis. The content of total bilirubin in Calculus Bovis from different sources varied greatly while being similar in in vitro cultured sam- pies. Bilirubin in covalent form occupied a great part in Calculus Bovis, and bilirubin in conjugated form hold a great proportionin in vitro cultured Calculus Bovis. CONCLUSION The contents or ratios of different types of bilirubin can be used as the quality control parameters of naturaland in vitro cultured Calculus Bovis. The established method is easy to operate with high effectiveness in quality control, suggesting a new way of identifying bezoar.

关 键 词：天然牛黄 体外培育牛黄 胆红素 含量测定 

分 类 号：R284[医药卫生—中药学]