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目的建立人血浆格列美脲及其代谢物同时检测的超高液相色谱-质谱方法。方法采用Waters超高液相质谱仪,以Acquity UPLC BEH C18柱(2.1 mm×50 mm,1.7μm)为色谱柱;以乙腈-0.1%甲酸为流动相,流速为0.4 mL·min-1,梯度洗脱;柱温为35℃。以格列本脲为内标,血浆经乙腈沉淀高速离心后检测。离子源为电喷雾离子源,正离子方式检测,扫描方式为多反应监测(MRM);用于定量分析的离子反应分别为:格列美脲m/z 491.2→126.15,代谢物M1 m/z 507.3→126.1,内标m/z494.2→369.0。结果血浆格列美脲浓度在2.5~400μg·L-1内线性关系良好(r=0.999 8);低、中、高3个浓度(5、50、300μg·L-1)的日内精密度RSD分别为7.23%、6.63%和4.69%,日间精密度RSD分别为8.66%、7.69%和5.76%;相对回收率分别为(102.57±5.43)%、(101.75±4.21)%和(101.67±3.81)%。代谢物M1浓度在0.5~100μg·L-1内线性关系良好(r=0.999 5);低、中、高3个浓度(1、10、75μg·L-1)的日内精密度RSD分别为8.74%、7.24%和4.33%,日间精密度RSD分别为10.68%、8.42%和5.64%;相对回收率分别为(103.12±6.52)%、(100.84±5.08)%和(99.01±3.22)%。结论该方法简便、快速、准确,适用于血浆格列美脲及其代谢物的定量检测、药动学和药物相互作用研究。
Objective To establish a method for the simultaneous determination of glimepiride and its metabolites in human plasma by ultra performance liquid chromatography-mass spectrometry. Methods A Waters UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) was used as the chromatographic column. The mobile phase was acetonitrile-0.1% formic acid and the flow rate was 0.4 mL · min-1. The gradient Elution; column temperature 35 ℃. Glibenclamide as internal standard, plasma was detected by high speed centrifugation after acetonitrile precipitation. Ion source for the electrospray ion source, positive ion detection, scanning mode for multiple reaction monitoring (MRM); for quantitative analysis of ion reaction were: glimepiride m / z 491.2 → 126.15, metabolites M1 m / z 507.3 → 126.1, internal standard m / z 494.2 → 369.0. Results The linearity of plasma glimepiride in 2.5 ~ 400μg · L-1 was good (r = 0.999 8). The intra-day precision of low, medium and high concentrations of 5,50,300μg · L-1 RSD (RSD) were 7.23%, 6.63% and 4.69%, respectively. The intraday precision RSD was 8.66%, 7.69% and 5.76% respectively; the relative recoveries were 102.57 ± 5.43%, 101.75 ± 4.21% and 101.67 ± 3.81% )%. The linear range of the metabolite M1 was 0.5-100 μg · L-1 (r = 0.999 5). The intra-day precision RSD of low, medium and high concentrations of 1, 10 and 75 μg · L-1 were 8.74 %, 7.24% and 4.33% respectively. The RSD of day precision was 10.68%, 8.42% and 5.64%, respectively. The relative recoveries were (103.12 ± 6.52)%, (100.84 ± 5.08)% and (99.01 ± 3.22)%, respectively. Conclusion The method is simple, rapid and accurate and is suitable for the quantitative detection, pharmacokinetics and drug interactions of glimepiride and its metabolites in plasma.