目的 建立一个灵敏度高,专属性好的高效液相色谱-质谱联用(HPLC-MS/MS)测定透析患者血浆中西那卡塞的浓度。方法 利用固相萃取(SPE)对血浆进行前处理,盐酸芬地林为内标(IS),采用内标法对药物进行定量。液相条件中色谱柱:Inertsil SIL-150A(2.1 mm×50 mm,5 μm),保护柱:Inertsil SIL-150A(3.0 mm×50 mm,5 μm),乙腈-水-甲酸(90∶10∶1=V∶V∶V)等度洗脱,流速0.35 mL·min-1,质谱采用电喷雾离子源(ESI),正离子模式下多离子反应监测(MRM)扫描,选用离子对为西那卡塞m/z 358.1→155.1,盐酸芬地林m/z 316.0→212.1,利用所建立的方法对口服25 mg西那卡塞(1片)的透析患者24 h内的血药浓度进行检测。结果 西那卡塞在0.1~50 ng·mL-1内呈良好的线性关系,准确度和精密度均在±15%以内,质控样品经长期存储或反复冻融以及样品处理后在进样器中的稳定性均在1.4%~3.9%内。西那卡塞的基质效应分别为(104.5±3.1)%、(105.6±4.3)%和(104.0±3.1)%,所有样品IS的基质效应为(106.4±3.0)%,符合生物样品定量分析方法验证指导原则的基本要求,能够满足对西那卡塞血药浓度的测定。结论 本实验建立了一个专属性强,灵敏度高,重现性好的HPLC-MS/MS方法,能够准确,快速的测定透析患者体内西那卡塞的血浆药物浓度。
Abstract
OBJECTIVE To establish a high performance liquid chromatography mass spectrometry (HPLC-MS/MS) method to determine the cinacalcet concentration in plasma of dialysis patients.METHODS Fendiline hydrochloride was selected as the internal standard (IS), in the meantime, plasma samples were extracted through the solid phase extraction (SPE), and the internal standard method was used to determine the concentration of cinacalcet. The analyte was separated by Inertsil SIL-150A (2.1 mm×50 mm, 5 μm) matched with guard column Inertsil SIL-150A (3.0 mm×50 mm,5 μm). The cinacalcet was eluted with acetonitrile, water and formic acid (90∶10∶1=V∶V∶V) at a rate of 0.35 mL·min-1. On MS, electrospray ionization (ESI) source with positive ion mode and multiply reaction monitoring (MRM) was used. The MRM transitions of cinacalcet and the IS were 358.1→155.1 and 316.0→212.1, respectively. The concentration in dialysis patients after 25 mg dose (one piece of tablet) within 24 h was determined by using the established HPLC-MS/MS method in this paper.RESULTS A good linearity was obtained in the range of 0.1-50 ng·mL-1 of cinacalcet, and the validated accuracy and precision all were within ±15%. Long-term, freeze-thaw and autoinjector stability of cinacalcet QC samples were all in the range of 1.4%-3.9%. The matrix of cinacalcet is (104.5±3.1)%, (105.6±4.3)% and (104.0±3.1)% respectively and the matrix of IS is (106.4±3.0)%. This validated method can meet the guidance for bioanalytical method validation.CONCLUSION The HPLC-MS/MS method is sensitive and specific enough to meet the quantitative requirements for determining cinacalcet in human plasma. It is rapid, accurate and in line with the guidance for bioanalytical method validation.
关键词
高效液相色谱-质谱联用 /
透析患者 /
西那卡塞 /
血药浓度
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Key words
HPLC-MS/MS /
dialysis patient /
cinacalcet /
plasma
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中图分类号：
R969.4
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参考文献
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脚注
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基金
复旦大学附属华山医院科研启动基金资助项目(2016QD06)
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