HPLC-MS同时测定人血浆中吡格列酮及其活性代谢产物的浓度

 目的建立高效液相色谱质谱联用测定人血浆中吡格列酮及其活性代谢产物M-Ⅲ(酮类衍生物)和M-Ⅳ(羟化衍生物)的方法。方法采用WatersXTerraTN C₁₈色谱柱(2.1mm×150mm,3.5μm),Phenomenex C₁₈保护柱,柱温50℃,流动相:乙腈-30mmol·L^-1醋酸铵溶液(含0.1%的甲酸和0.05%的三氟醋酸)(35:65),流速0.22mL·min^-1。质谱采用电喷雾电离源正离子模式(ESI⁺),选择性监测质荷比(m/z)357.4(吡格列酮),358.2(内标),371.5(M-Ⅲ),373.2(M-Ⅳ)的准分子分子离子峰;血浆样品经盐酸酸化后,采用叔丁基甲醚-氯仿提取,内标法定量。结果吡格列酮,M-Ⅲ和M-Ⅳ分别在11.16～1748.60,3.33～520.50,5.00～687.50ng·mL^-1内线性良好(r≥0.9997),最低检测浓度分别为2.90,1.10,1.20ng·mL^-1,3个化合物的方法回收率均在90%-110%范围内,日间和日内RSD皆小于15%。结论本方法操作简单,灵敏准确,稳定性好,适用于临床吡格列酮治疗药物监测,药动学及其药物相互作用的研究。

Simultaneous determination of pioglitazone and its two active metabolites in human plasma by HPLC-MS

OBJECTIVE To establish a HPLC-MS method for simultaneous determinating of pioglitazone and its two active metabolites: M-Ⅲ(keto-derivative) and M-Ⅳ(hydroxy-derivative) in human plasma.METHOD The separation was performed on a Waters XTerraTM C₁₈ column(2.1 mm×150 mm,3.5 μm) with guard column Phenomenex C₁₈. The column temperature was 50℃. The mobile phase consisted of acetonitrile-30 mmol·L^-1 ammonium acetate solution(added with 0.1% formic acid and 0.05% trifluoroacetic acid) (35:65), with a flow rate of 0.22 mL·min^-1. The compound was inoized in the electrospray ionization(ESI) ion source of the mass spetrometer and selected ion mass spectral(m/z) 357.4(PIO), 358.2(is), 371.5(M-Ⅲ), 373.2(M-Ⅳ)to quantify. Human plasma samples were extracted with 1:2 chlor-form:methyl t-butyl ether after acidification RESULTS The linear ranges were 11.16～1 748.60 ng·mL^-1 for pioglitazone,3.33～520.50 ng·mL^-1 for M-Ⅲ and 5.00～687.50 ng2mL^-1 for M-Ⅳ(r≥0.9997), and their detect limits were 2.90, 1.10, 1.20ng·mL^-1. Recoveries were within 90%～110% , and intra-and inter-day RSDs were all less than 15%.CONCLUSION The method is found to be sensitive, rapid and accurate, and has been applied successfully to sample analysis for clinical study of pioglitazone pharmacokinetics and drug interaction.