摘要
目的:建立LC-MS/MS法同时测定大鼠血浆中黄芩素及其代谢物黄芩苷浓度。方法:色谱分离采用C18色谱柱,流动相为甲醇-0.2%甲酸(74∶26),流速1 mL/min。血浆中加入内标木犀草素,甲醇沉淀蛋白,离心后上清液进行LC-MS/MS分析。检测离子:黄芩素[M+H]+270.9→122.8,黄芩苷[M+H]+447.0→271.3,内标[M+H]+286.9→153.0。结果:黄芩苷、黄芩素在10-2 000 ng/mL范围内,线性关系良好,平均回收率大于75%,日内和日间的RSD小于15%。结论:建立的LC-MS/MS法符合生物样品分析的要求,可用于研究大鼠静注黄芩素后,血浆中黄芩素、黄芩苷的同时测定。
Aim： To develop a rapid and sensitive liquid chromatography-tandem mass spectrometry （LC-MS/MS） method for the simultaneous determination of baicalein and baicalin in the rat＇s plasma. Methods： Plasma samples, into which luteolin was added as internal standard, were deproteinized by methanol and centrifuged for sample clean-up. The yielded supernatant was then evaporated to dryness, and the residue was reconstituted in 100 μL of mobile phase and centrifuged. An aliquot of 20 μL was injected into HPLC column. Chromatographic separation was achieved on a C18 column and the mobile phase consisted of methanol and 0.2% formic acid （74：26） pumped at a flow rate of 1.0 mL/min. The mass spectrometer was performed in the selected reaction monitoring （SRM） mode and the transitions selected for quantitation were [M＋H]^＋ 270.9→122.8 for baicalein, [M＋H]^＋ 447.0→271.3 for baicalin and [M＋H]^＋286.9→153.0 for luteolin, respectively. Results： A liner calibration curve for the assay of baicalein and baicalin in the rat＇s plasma was validated in the range of 10 - 2 000 ng/mL with the recoveries of at least 75% and intra-day and inter-day RSD of 15 %. Conclusion： The method can be used for the simultaneous determination of baicalein and baicalin in the rat＇s plasma.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2007年第2期129-132,共4页
Journal of China Pharmaceutical University
基金
江苏新药创制发展基金资助项目(No.06060)~~
