天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (10): 1096-1101.doi: 10.11958/20180795

• 新技术交流 • 上一篇    下一篇

LC-MS/MS法分析人血浆中西格列汀浓度及其应用研究

刘珊珊 1,2，夏媛媛 2，刘静媛 2，耿雅杰 2，魏广力 2，司端运 2△   

  1. 基金项目：国家自然科学基金青年科学基金资助项目（81503154） 作者单位：1天津医科大学研究生院（邮编300070）；2天津药物研究院新药安全评价有限公司 作者简介：刘珊珊（1993），女，硕士在读，主要从事释药技术与药代动力学研究 △通讯作者 E-mail: sidy@tjpr.com
  • 收稿日期:2018-05-16 修回日期:2018-07-26 出版日期:2018-10-15 发布日期:2018-11-09
  • 通讯作者: 刘珊珊 E-mail:1029190268@qq.com
  • 基金资助:
    国家自然科学基金青年科学基金

Analytical methods and application of LC-MS/MS for quantification of sitagliptin in human plasma

LIU Shan-shan1,2, XIA Yuan-yuan2, LIU Jing-yuan2, GENG Ya-jie2, WEI Guang-li2, SI Duan-yun2△   

  1. 1 Tianjin Medical University Graduate School, Tianjin 300070, China; 2 State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research △Corresponding Author E-mail: sidy@tjpr.com
  • Received:2018-05-16 Revised:2018-07-26 Published:2018-10-15 Online:2018-11-09
  • Contact: LIU Shanshan E-mail:1029190268@qq.com

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摘要： 摘要：目的 建立一种测定人血浆中西格列汀浓度的液相色谱-串联质谱（LC-MS/MS）分析方法，并将该方法应 用于西格列汀在人体内的药代动力学研究。方法 以西格列汀-d4为内标，血浆样品经CleanertPPT沉淀板沉淀后， 通过Diamonsil C18色谱柱（100 mm×4.6 mm，5 μm）进行分离，使用甲醇-10 mmol/L甲酸铵水溶液（含10%甲醇，0.1% 甲酸）作为流动相，进行梯度洗脱，流速为0.5 mL/min。通过电喷雾电离源（ESI），以多反应监测（MRM）模式进行正离 子检测。从选择性、残留、线性范围与定量下限、精密度与准确度、基质效应和回收率、稳定性方面进行方法学验证。 同时考察健康人口服西格列汀片100 mg后的主要药代动力学参数。结果 西格列汀、西格列汀-d4的MRM离子对 分别为 m/z 408.0→235.2、m/z 412.1→239.0。人血浆中西格列汀在 0.5～1 000 μg/L 浓度范围内线性关系良好（R2> 0.99），定量下限为0.5 μg/L；定量下限和质控样品的批内、批间精密度（RSD）在0.83%~12.80%之间，准确度（RE）在± 10.0%以内。健康人口服西格列汀片100 mg后主要药代动力学参数：达峰时间（Tmax）、达峰浓度（Cmax）、、生物利用度 （AUC）、半衰期（T1/2）分别为（2.44±1.29）h、（375±138）μg/L、（2 915±585）h·μg/L、（11.10±2.41）h。结论 本LC-MS/ MS分析方法敏感度高且样品处理方法简单快速，满足生物分析的法规要求，可应用于人体内西格列汀的药代动力学 研究。

关键词: 二肽基肽酶Ⅳ抑制剂, 药代动力学, 西格列汀, 高效液相, 串联质谱法

Abstract: Abstract: Objective To develop a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method for quantification of sitagliptin in human plasma, and to apply it for the pharmacokinetics study of sitagliptin in human. Methods The plasma sample was precipitated by CleanertPPT and sitagliptin-d4 was used as the internal standard (IS). Chromatographic separation was performed on a Diamonsil C18 column (100 mm×4.6 mm, 5 μm). Mobile phase consisted of methanol and a mixture of 10 mmol/L ammonium formate-methanol-formic acid (90∶10∶0.1) at a flow rate of 0.5 mL/min. The validation was performed in terms of selectivity, residue, linear range and lower limit of quantitation, precision and accuracy, matrix effects and recovery, and stability. The main pharmacokinetic parameters (Tmax, Cmax, AUC0-t and T1/2) were observed in healthy people after oral administration of sitagliptin tablets. Results Sitagliptin and sitagliptin-d4 (IS) were ionized with an ESI source and operated in positive ion mode. The detected ions were m/z 408.0→235.2 (sitagliptin) and m/z 412.1→239.0 (sitagliptin-d4). This validated LC-MS/MS method yielded a good linearity over the range of 0.5-1 000 μg/L (R2=0.99), and the lower limit of quantitation (LLOQ) was 0.5 μg/L. The intra and inter-assay precisions (RSD) were within the range of 0.83%~12.80%, and the accuracy (RE) was less than ± 10.0%. The pharmacokinetic parameters, Tmax, Cmax, AUC 0-t and T1/2 were (2.44±1.29) h, (375±138) μg/L, (2 915±585) h·μg/L and (11.10±2.41) h. Conclusion This LC-MS / MS method is simple, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.

Key words: dipeptidyl-peptidase Ⅳ inhibitors, pharmacokinetics, Sitagliptin, liquid chromatography-tandem mass spectrometry, tandem mass spectrometry