Simultaneous determination of piperaquine and its N‐oxidated metabolite in rat plasma using LC‐MS/MS

Abstract

A sensitive and efficient liquid chromatography tandem mass spectrometry method was developed and validated for the simultaneous determination of piperaquine (PQ) and its N-oxidated metabolite (PQ-M) in plasma. A simple protein precipitation procedure was used for sample preparation. Adequate chromatographic retention was achieved on a C column under gradient elution with acetonitrile and 2 mm aqueous ammonium acetate containing 0.15% formic acid and 0.05% trifluoroacetic acid. A triple-quadrupole mass spectrometer equipped with an electrospray source was set up in the positive ion mode and multiple reaction monitoring mode. The method was linear in the range of 2.0-400.0 ng/mL for PQ and 1.0-50.0 ng/mL for PQ-M with suitable accuracy, precision and extraction recovery. The lower limits of detection (LLOD) were established at 0.4 and 0.2 ng/mL for PQ and PQ-M, respectively, using 40 μL of plasma sample. The matrix effect was negligible under the current conditions. No effect was found for co-administrated artemisinin drugs or hemolysis on the quantification of PQ and PQ-M. Stability testing showed that two analytes remained stable under all relevant analytical conditions. The validated method was successfully applied to a pharmacokinetic study performed in rats after a single oral administration of PQ (60 mg/kg).

Determination of piperaquine concentration in human plasma and the correlation of capillary versus venous plasma concentrations

“…Due to this property, peak tailing and carryover peaks were often observed during the analysis of PQ. The majority of published LC-MS/MS methods utilized acidic mobile phase solvents [3,[9][10][11][12], while two methods used basic mobile phase solvents [8,23] (Table 5). Carryover peaks were observed in nearly all published LC-MS/MS methods.…”

“…Carryover could be due to the column (tailing peak), precolumn tubing, or autosampler. Numerous methods have been published for the quantification of PQ in plasma, including high performance liquid chromatography-ultraviolet/visible spectrophotometry (HPLC-UV) [5][6][7] and liquid chromatography tandem mass spectrometry (LC-MS/MS) [8][9][10][11][12]. Carryover was observed in nearly all LC-MS/ MS methods.…”

Metabolism of Piperaquine to Its Antiplasmodial Metabolites and Their Pharmacokinetic Profiles in Healthy Volunteers

“…A liquid chromatography-mass spectrometry method was applied for quantification of PQ and its metabolite on an API5500 Q-Trap triple quadrupole mass spectrometer (AB Sciex, Concord, Ontario, Canada) equipped with a turbo ion spray source. The sample preparation and chromatographic and mass spectrometric conditions have been described in a previous report (15), and the analytical method was fully validated. Representative chromatograms for quantification of PQ and its metabolites are shown in Fig.…”

“…The antimalarial efficacy of the other two metabolites (M1 and M2) remains unknown. The pharmacokinetic profiles of PQ and its metabolites could be of great importance in determining the clinical efficacy and safety outcomes of PQ; however, related information was limited only to the pharmacokinetics of the parent drug PQ and its metabolites in rats (15) or in only one human subject (16).…”