化学
色谱法
三氟乙酸
乙胺丁醇
蛋白质沉淀
吡嗪酰胺
质谱法
串联质谱法
液相色谱-质谱法
样品制备
定量分析(化学)
高效液相色谱法
利福平
生物化学
抗生素
作者
Zhaoning Gong,Yousef J. Basir,David C. Chu,Melanie McCort-Tipton
标识
DOI:10.1016/j.jchromb.2009.04.023
摘要
Ethambutol and pyrazinamide are two first-line anti-tuberculosis drugs. Though they are normally combined for the treatment, their highly different polarity complicates simultaneous liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis of these two drugs in human plasma with decent peak shape and retention. Here we report a rapid and robust LC/MS/MS method for the simultaneous determination of these two drugs in human plasma. Human plasma samples, together with the isotopically labeled internal standards were extracted using protein precipitation, and then separated on a Chromolith SpeedROD RP-18e column and detected with mass spectrometry. The mobile phase is 0.1% trifluoroacetic acid in water and 0.1% trifluoroacetic acid in methanol. Addition of trifluoroacetic acid in the mobile phases was found to be able to improve peak shape as well as to increase the retention of ethambutol, thus being able to analyze these two drugs at the same time with both drugs having decent peak shape and enough retention on a C18 column. An atmospheric pressure chemical ionization interface was chosen to reduce ion suppression from sample matrix components and provide high sensitivity. The standard curve range was 10.0-5000 ng/mL for ethambutol and 50.0-25,000 ng/mL for pyrazinamide using a plasma sample volume of 50.0 microL. This method has a very short run time of 3.8 min. The method has been fully validated, and <15% relative standard deviation was obtained for both analytes.
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