LC-MS/MS法同时测定人血浆中氟吡汀及其代谢产物D-13223的浓度: 马来酸氟吡汀胶囊在健康男性志愿者中的应用

doi:10.5246/jcps.2021.10.070

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (10): 822-830.DOI: 10.5246/jcps.2021.10.070

LC-MS/MS法同时测定人血浆中氟吡汀及其代谢产物D-13223的浓度: 马来酸氟吡汀胶囊在健康男性志愿者中的应用

刘艳芳¹, 张嘉珊², 唐云彪¹^,^*(), 霍花¹^,^*()

1. 1. 北部战区总医院临床药学技术中心, 辽宁沈阳 110840
2. 2. 沈阳药科大学生命科学与生物制药学院临床药学专业, 辽宁沈阳 110840

收稿日期:2021-04-23 修回日期:2021-05-24 接受日期:2021-06-02 出版日期:2021-10-24 发布日期:2021-10-24
通讯作者: 唐云彪, 霍花
作者简介:
+ Tel.: +86-17790990199, E-mail: hh_602@sina.com
+ Tel.: +86-13372828008, E-mail: tangyb99@163.com

Quantification of flupirtine and its active metabolite D-13223 in human plasma by LC-MS/MS: application to a clinical trial of flupirtine maleate capsules in healthy male Chinese volunteers

Yanfang Liu¹, Jiashan Zhang², Yunbiao Tang¹^,^*(), Hua Huo¹^,^*()

1 Technical Center for Clinical Pharmacy, General Hospital of Northern Theater Command, Shenyang 110840, China
2 Department of Clinical Pharmacy, School of Life Sciences and Biopharmaceutical, Shenyang Pharmaceutical University, Shenyang 110840, China

Received:2021-04-23 Revised:2021-05-24 Accepted:2021-06-02 Online:2021-10-24 Published:2021-10-24
Contact: Yunbiao Tang, Hua Huo

摘要/Abstract

摘要：

建立一种简便、灵敏、准确的液相色谱-串联质谱法以同时测定人血浆中氟吡汀及其活性代谢物D-13223浓度。本文采用三重四级杆液质联用仪, 电喷雾电离源(ESI), 多反应离子监测(MRM)的正离子模式进行检测, 以C18柱为色谱柱, 乙腈–水–氨(55:45:0.1, v/v/v)为流动相, 液液萃取法提取血浆样品中氟哌汀、D-13223及稳定同位素内标以进行色谱分析。氟吡汀在10.00–2000.00 ng/mL浓度范围内线性关系良好, D-13223在2.00–400.00 ng/mL浓度范围内线性关系良好。氟吡汀及D-13223质量控制(QC)样品的批内和批间精密度均小于9.26%, 准确度范围–5.80%–3.31%。氟吡汀、D-13223及内标的提取回收率均在88.3%–97.2%之间。本法已成功应用于健康男性志愿者口服100 mg马来酸氟吡汀胶囊后血浆中氟吡汀及其活性代谢物D-13223的药代动力学研究。

关键词: 氟吡汀, D-13223, LC-MS/MS, 定量测定

Abstract:

In the present study, we developed a simple, sensitive, precise, and accurate liquid chromatography-tandem mass spectrometric (LC-MS/MS) method and validated such approach for simultaneous determination of flupirtine and its active metabolite D-13223 in human plasma. The flupirtine, D-13223, and stable isotope internal standard (IS) were extracted from plasma samples by liquid-liquid extraction and chromatographed on a C18 column with a mobile phase consisting of acetonitrile–water–ammonia (55:45:0.1, v/v/v) at a flow rate of 0.25 mL/min. Detection was performed on a triple quadrupole tandem mass spectrometer with an electrospray ionization source (ESI) by multiple reaction monitoring (MRM) in positive ion mode. The linear calibration curves were obtained within the concentration range of 10.00–2000.00 ng/mL for flupirtine and 2.00–400.00 ng/mL for D-13223. The intra- and inter-run RSD, calculated from quality control (QC) samples, was less than 9.26% for flupirtine and D-13223. The accuracy was –5.80%–3.31% for flupirtine and D-13223. The extraction recoveries of flupirtine, D-13223, and their IS were all between 88.3%–97.2%. The method was successfully applied to investigate the pharmacokinetic profiles of flupirtine and its active metabolite D-13223 in human plasma following peroral administration of 100 mg flupirtine maleate capsules in healthy male Chinese volunteers.

Key words: Flupirtine, D-13223, LC-MS/MS, Clinical trial

Supporting:

刘艳芳, 张嘉珊, 唐云彪, 霍花. LC-MS/MS法同时测定人血浆中氟吡汀及其代谢产物D-13223的浓度: 马来酸氟吡汀胶囊在健康男性志愿者中的应用[J]. 中国药学(英文版), 2021, 30(10): 822-830.

Yanfang Liu, Jiashan Zhang, Yunbiao Tang, Hua Huo. Quantification of flupirtine and its active metabolite D-13223 in human plasma by LC-MS/MS: application to a clinical trial of flupirtine maleate capsules in healthy male Chinese volunteers[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(10): 822-830.

图/表 7

Figure 1. Flupirtine (1) and D-13223 (2).

Table 1. The mass spectral parameters of analytes and IS.

Figure 2. The representative chromatograms of blank plasma (A), blank plasma added to flupirtine, D-13223, the IS F-d5, D-d4 (B), and plasma sample obtained from subject 1 after oral administration of katadolon 1.5 h (C). 1. Flupirtine; 2. D-13223; 3. F-d5; 4. D-d4.

Table 2. Precision and accuracy for the analysis of flupirtine and D-13223 in QC samples (n = 3 runs, six replicates per day) in human plasma.

Table 3. The average extraction recovery for flupirtine, D-13223, and IS.

Table 4. Stability of flupirtine and D-13223 in human plasma (n = 3).

Figure 3. Mean plasma concentration-time profiles of flupirtine and D-13223 under fasting (A) and fed (B) conditions after an oral dose of 100 mg test capsule and reference capsule.

参考文献 24

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LC-MS/MS法同时测定人血浆中氟吡汀及其代谢产物D-13223的浓度: 马来酸氟吡汀胶囊在健康男性志愿者中的应用

Quantification of flupirtine and its active metabolite D-13223 in human plasma by LC-MS/MS: application to a clinical trial of flupirtine maleate capsules in healthy male Chinese volunteers

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