高效液相-质谱联用测定阿奇霉素的血浆浓度

Determination of azithromycin in human plasma by liquid chromatography mass spectrometry

目的:本实验建立了高效液相-质谱联用(HPLC-MS/MS)法测定正常人血浆中的阿奇霉素浓度。方法:血浆中加入罗红霉素作为内标,以碱化的醋酸乙酯提取,进行HPLC-MS/MS的测定。采用Hanbon Lichrospher C18柱(2.1mm×100mm,5μm),柱温30℃,流动相为乙睛-醋酸铵缓冲液(0.2%醋酸+0.1%醋酸铵溶液)(60∶40),流速0.25mL·min-1;质谱采用选择性离子检测方法。结果:阿奇霉素的线性范围为1.0～1000mg·L-1,最低检测质量浓度为1.0μg·L-1。回收率及精密度的测定均符合生物样品分析的要求。结论:该方法操作简便,灵敏度高,专属性强,可用于测定阿齐霉素的血药浓度。

ORJECTIVE To established an HPLC MS/MS method to determine of azithromycin in human plasma, for pharmacokinetic study. METHODS The drug was extracted by diethyl ether from human plasma with the clarithromycin as internal standard. The evaporated samples were dissolved in mobile phase and separated by Hanbon Lichrospher C18 （100 mm × 2. 1 mm, 5/am） column. The eluate from the ttPLC column was plumbed directly into ESI probe with the mobile phase containing （0. 1% ammonium acetate-0. 2% acetonitrile）-acetonitrile （40：60） at flow rate of 0. 25 mL.min-1. Analysis in the mass spec- trometer was operated in the selection monitoring mode. RESULTS The method demonstrated that good linearity ranged from 1 to l 000μg.L-1. The limit of quantification for azithromycin in plasma was μg.L-1 with good accuracy and precision, and so was the extraction recovery. CONCLUSION The method has been successfully applised to the bioequivalence study of azithro- mycin for 20 healthy human plasma.