高效液相色谱-串联质谱法测定人血浆中异烟肼、利福平及其在健康男性体内的药物代谢动力学研究
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上海市徐汇区中心医院课题(2018XHYY-22)。


Levels of Isoniazid and Rifampicin levels in human plasma determined by high performance liquid chromatography-tandem mass spectrometry and their pharmacokinetics in healthy Chinese males
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    摘要:

    目的?建立可靠的测定人血浆中异烟肼(INH)和利福平(RFP)浓度的高效液相色谱-串联质谱法,用于研究健康男性体内的药物代谢动力学特征。方法?以稳定同位素作为内标,血浆样品经甲醇沉淀蛋白质处理后检测。INH采用Atlantis HILIC(50 mm×2.1 mm,3 μm)色谱柱,以水和5 mmol/L甲酸铵90%乙腈水溶液(均含0.01%甲酸)为流动相;RFP 采用Syncronis C18(50 mm×2.1 mm,5 μm)色谱柱,以乙腈和50%乙腈水溶液为流动相;二者均为梯度洗脱。使用电喷雾离子源,正离子多反应监测方式进行检测。结果?INH、RFP 分别在5~5 000 ng/mL、10~10 000 ng/mL范围内线性关系良好(r>0.99)。方法专一性强,精密度以变异系数表示,均小于7%,无明显基质效应。结论?本方法操作简便,特异性高,可用于人体药物代谢动力学及治疗药物监测等研究。

    Abstract:

    Objective?To establish an accurate,reliable method for determination of Isoniazid(INH)and Rifampicin(RFP)in human plasma by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS),and investigate the pharmacokinetics characteristics in healthy Chinese male volunteers.Methods?By using stable isotopes as internal standards,the plasma samples were pretreated by precipitation with methanol,and then diluted and analyzed.Gradient elution was performed for both INH and RFP.INH was separated on an Atlantis HILIC by a HILIC Silica(50 mm×2.1 mm,3 μm)with 5 mmol/L ammonium formate,90% acetonitrile aqueous solution containing 0.01% formic acid as mobile phase.RFP were separated by a Syncronis C18(50 mm×2.1 mm,5 μm)with acetonitrile-50% acetonitrile aqueous solution as mobile phase.Electrospray ionization source was applied for determination in positive ion multi-reaction monitoring mode.Results?The linear ranges of INH and RFP were 5 to 5 000 ng/mL and 10 to 10 000 ng/mL respectively with fine correlation(r>0.99).The coefficient of variation of precision was less than 7%.No obvious matrix effect was found.Conclusion?The established method should be simple,accurate and specific and may be used in the study for human pharmacokinetics and drug monitoring in therapy.

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张美微,陆优丽,忻亮,李水军,刘罡一,董春霞,张梦琪,梁立宇,缪怡,贾晶莹.高效液相色谱-串联质谱法测定人血浆中异烟肼、利福平及其在健康男性体内的药物代谢动力学研究[J].临床检验杂志,2021,39(7):489-494

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  • 收稿日期:2020-11-11
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  • 在线发布日期: 2021-09-09
  • 出版日期: 2021-07-28
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