Quantification of cefaclor in human plasma using SIL‐IS LC‐ESI‐MS/MS for pharmacokinetics study in healthy Chinese volunteers

Abstract

A steady, high‐efficiency, and precise liquid chromatography‐electrospray ionization‐tandem mass spectrometry method was established and validated using cefaclor‐d5 as the stable isotope‐labeled internal standard for quantification of cefaclor in human plasma. One‐step protein precipitation was applied to extract human plasma samples using methanol as precipitant. An Ultimate XB C18 column (2.1 × 50.0 mm, 5.0 μm) was used to achieve chromatographic separation. Mobile phases of gradient elution were an aqueous solution containing 0.1% formic acid (mobile phase A) and an acetonitrile solution containing 0.1% formic acid (mobile phase B). Electrospray ionization in positive‐ion mode was applied to detect under multiple reaction monitoring mode. Target fragment ion pairs of cefaclor and stable isotope‐labeled internal standard, respectively, were m/z 368.2 → 191.1 and m/z 373.2 → 196.1. Linear range of this method was between 20.0 and 10,000.0 ng/ml, with coefficient of determination (R2) >0.9900. Seven concentrations of quality control samples were used: 20.0 ng/ml (lower limit of quantitation), 60.0 ng/ml (low quality control), 650 ng/ml (middle quality control), 5000 ng/ml (arithmetic average middle quality control [AMQC]), 7500 ng/ml (high quality control), 10,000 ng/ml (upper limit of quantification), and 40,000 ng/ml (dilution quality control [DQC]). The method was validated for selectivity, lower limit of quantitation, linearity, accuracy, precision, recovery, matrix effect, dilution reliability, stability, carryover, and incurred sample reanalysis. This stable isotope‐labeled internal standard liquid chromatography‐electrospray ionization‐tandem mass spectrometry approach has been successfully applied to study the pharmacokinetics of cefaclor dry suspension among healthy Chinese volunteers.

LC–MS/MS method for quick detection of vonoprazan fumarate in human plasma: Development, validation and its application to a bioequivalence study

“…SIL‐IS has similar physicochemical properties and chromatographic behavior as the analyte while it could be distinguished from the analyte on the mass spectrum (Liu et al, 2019), as well as being able to correct the influences of the matrix effect on the analytical results and improve the extraction recovery and ionization efficiency (Wang et al, 2022; Zhan et al, 2021). Compared with using ammonium formate as the mobile phase (Yoneyama et al, 2016), formic acid had good compatibility with mass spectrometry (MS) detection and did not produce salt deposition in the MS source, which could affect the stability and sensitivity of MS signals (Chen et al, 2023); therefore, formic acid was chosen as the mobile phase. Acetonitrile as the precipitant showed a higher precipitation efficiency than methanol with recovery between 97.0 and 100.8%, and the precipitation products of acetonitrile were more easily separated than methanol without interference of the precipitates in subsequent analysis.…”