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

doi:10.5246/jcps.2021.10.070

Abstract

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:

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.

Figures/Tables 7

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

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).

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