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<p>A robust and reproducible LC–MS/MS method was developed and optimized for the simultaneous quantification of artemether (ART) and lumefantrine (LUF) in rat plasma, employing carbamazepine as an internal standard (IS). The mass spectrometer operated in positive ionization mode with electrospray ionization (ESI), yielding product ions at m/z 163.05(ART), m/z 511.95 (LUF), and m/z 194.1 (IS). Chromatographic separation was achieved using a Waters™ X Select C18 column (100&#xa0;mm × 4.6&#xa0;mm, 2.5&#xa0;µm) with a mobile phase of 0.2&#xa0;% formic acid in water: methanol (10:90) at a flow rate of 0.5&#xa0;mL/min. Retention times for ART, LUF, and IS were 4.30&#xa0;min, 1.60&#xa0;min, and 2.46&#xa0;min, respectively, within a 6-min runtime. The liquid–liquid extraction (LLE) method utilized ethyl acetate with acidified methanol for optimal analyte recovery, achieving mean recoveries of 92.12&#xa0;% for ART and 91.07% for LUF. Calibration curves were linear over the range of 5–1000&#xa0;ng/mL with regression coefficients of ≥ 0.9992. The method demonstrated high precision (%CV: 0.16–1.89) and accuracy (98.24–106.42&#xa0;%) at the lower limit of quantification (LLOQ: 5&#xa0;ng/mL). The method exhibited minimal matrix effects, with stability tests confirming analyte integrity under benchtop and freeze–thaw conditions. To the best of our knowledge, this is the first validated LC-MS/MS method developed for the simultaneous quantification of ART and LUF in rat plasma for preclinical pharmacokinetic studies, providing a novel, sensitive, and cost-effective analytical tool for routine high-throughput analysis.</p>

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Simultaneous Quantification of Artemether and Lumefantrine in Rat Plasma Using a Validated LC-MS/MS Method

  • 期刊论文

  • 发表日期：

    2025年7月10日

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  • Ripal Shah,
  • Tejal Soni,
  • Dinal Patel,
  • Jenee Christian,
  • Mansi Dholakia,
  • Mehul Patel,
  • Kishor Sorathia

摘要

A robust and reproducible LC–MS/MS method was developed and optimized for the simultaneous quantification of artemether (ART) and lumefantrine (LUF) in rat plasma, employing carbamazepine as an internal standard (IS). The mass spectrometer operated in positive ionization mode with electrospray ionization (ESI), yielding product ions at m/z 163.05(ART), m/z 511.95 (LUF), and m/z 194.1 (IS). Chromatographic separation was achieved using a Waters™ X Select C18 column (100 mm × 4.6 mm, 2.5 µm) with a mobile phase of 0.2 % formic acid in water: methanol (10:90) at a flow rate of 0.5 mL/min. Retention times for ART, LUF, and IS were 4.30 min, 1.60 min, and 2.46 min, respectively, within a 6-min runtime. The liquid–liquid extraction (LLE) method utilized ethyl acetate with acidified methanol for optimal analyte recovery, achieving mean recoveries of 92.12 % for ART and 91.07% for LUF. Calibration curves were linear over the range of 5–1000 ng/mL with regression coefficients of ≥ 0.9992. The method demonstrated high precision (%CV: 0.16–1.89) and accuracy (98.24–106.42 %) at the lower limit of quantification (LLOQ: 5 ng/mL). The method exhibited minimal matrix effects, with stability tests confirming analyte integrity under benchtop and freeze–thaw conditions. To the best of our knowledge, this is the first validated LC-MS/MS method developed for the simultaneous quantification of ART and LUF in rat plasma for preclinical pharmacokinetic studies, providing a novel, sensitive, and cost-effective analytical tool for routine high-throughput analysis.