反相离子对色谱法同时测定大鼠血浆中五种大黄蒽醌的方法学研究  

Study on reversed phase ion-pair HPLC determination method of rhubarb free anthraquinones in rats plasma

作　　者：王平[1] 缪舒益[1] 孟宪丽[1] 杨永茂[1] 曾勇[1] 

机构地区：[1]成都中医药大学药学院,成都611730

出　　处：《中药药理与临床》2009年第5期47-49,共3页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金资助项目(No:30772764);四川省教育厅自然科学青年基金(No:2006B021)。

摘　　要：目的：建立大鼠血浆中5种大黄蒽醌同时测定的反相离子对HPLC定量方法。方法：以1,8-二羟基蒽醌为内标,血浆预处理采用4倍样本量的甲醇沉淀,上清液在37℃水浴中用N2柔和吹干后流动相复溶进样。分析柱为Scienhome kromasil ODS-1（C18,250mm×4.6 mm,5μm）;流动相为：A：甲醇,B：水相（其中含20mM磷酸二氢钾,含6mM十六烷基三甲基溴化铵,pH=4.86）,A：B=（80：20）;流速：1.00 ml/min;检测波长：254nm;柱温：30℃;内标法定量。结果：本法测得血浆中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的线性范围分别为0.061~29.687μg/ml、0.050~24.450μg/ml、0.062~30.300μg/ml、0.170~33.100μg/ml、0.074~14.494μg/ml,r值均大于0.999,各成分提取回收率均大于69%,准确度93~101%之间,日内与日间精密度RSD均小于10%。结论：本测定方法各成分专属性强,分离度好,分析时长适宜,样本需求量少,灵敏度高,准确、精密、稳定,适合大黄游离蒽醌的多成分药物动力学研究。Objective：A reversed phase ion-pair HPLC method was developed for the simultaneous determination of five Rhubarb Free Anthraquinones in rats plasma.Methods： 1,8-dihydroxyanthraquinone was used as the internal standard（IS）,The plasma samples were pretreated including following steps：（1） to extract rhubarb anthraquinones with methanol,then the methanol layer was evaporated to dryness with N2 in 37℃ water bath.（2） to reconstitute the residue with the mobile phase and inject to HPLC for analysis.Rhubarb anthraquinones and IS were separated on Scienhome kromasil C18 column（250mm×4.6 mm,5μm）;cetyl tetrabutyl ammonium bromidethe（CTAB） was used as the ion-pair reagent and the mobile phase was made up of methanol and water（80：20,v/v;water phrase include 20mM KH2PO4 and 6mM CTAB,pH=4.86）,The ultraviolet detection was 254 nm.Results： Under the separation condition,aloe-emodin,rhein,emodin,chrysophanol,phycsion and IS in plasma were separated.The assay was linear over the range 0.061～29.687μg/ml,0.050～24.450μg/ml,0.062～30.300μg/ml,0.170～33.100μg/ml,0.074～14.494μg/ml（all r0.999） for aloe-emodin,rhein,emodin,chrysophanol,phycsion,respectively.The average recoveries were all above 69%.The precision is between 93~101%,their RSDs of intra-day and inter-day precision were less than 10%.Conclusion： The method is simple,sensitive,accurate,precise and stable,and suitable for the study of Rhubarb anthraquinones in vivo.

关 键 词：大黄蒽醌 反相离子对 HPLC 血浆 

分 类 号：R285.5[医药卫生—中药学]