Quantitative analysis of acetaminophen and its six metabolites in rat plasma using liquid chromatography/tandem mass spectrometry

Abstract

Acetaminophen (APAP) is a widely used analgesic and antipyretic drug. It is mainly metabolized by phase 1 and 2 reactions in the liver, and thus it could be involved in many drug-drug interactions. Therefore, the study of APAP metabolism is important in toxicological and pharmacokinetic studies. The objective of this study was to develop a rapid and sensitive method for the determination of APAP and its six metabolites in rat plasma for the pharmacokinetic studies. APAP and its metabolites were separated through a Capcell Pak MGII C(18) column and quantitated with a 16 min run in a triple-quadruple mass spectrometer. The mobile phases were composed of 0.1% formic acid in either 95% water or 95% acetonitrile and analysis was performed twice in positive and negative modes. Validations such as accuracy, precision, recovery, matrix effect and stability were found to be within acceptance criteria of validation guidelines, indicating that the assay was applicable to the determination of the plasma concentrations of drug and its six metabolites. In conclusion, we developed an LC-MS/MS method for the quantitative analysis of APAP and its six metabolites in rat plasma, and this method appears to be useful for pharmacokinetic/toxicokinetic studies of APAP and its metabolites in rats.

Acetaminophen Prodrug: Microwave-Assisted Synthesis andin vitroMetabolism Evaluation by Mass Spectrometry

“…An et al 33 and Ma et al 34 have detected the paracetamol reactive metabolite 3 by trapping this short half-live intermediate with glutathione. Interestingly, when the microsomal reaction of 2 was performed in the presence of this trapping agent neither the expected adduct nor different derivatives were detected by ion trap tandem mass spectrometry, which indicates that the acetaminophen first phase metabolism did not occur in the microsomal reaction studied.…”

Spectrophotometric methods for the simultaneous determination of paracetamol and dantrolene sodium in pharmaceutical dosage form

“…Several methods have been reported for the determination of PAR either alone [2][3][4], or with its metabolites in biological fluids [5][6][7], or in combination with other drugs [8][9][10][11][12]. DAS was also determined either alone [13] or in presence of its metabolites and impurities [14,15] or in biological fluids [16,17].…”

Simultaneous quantification of acetaminophen and structurally related compounds in human serum and plasma

“…[19][20][21][22][23][24][25] Various other analytical methods have also been applied to detect APAP and its metabolites, such as high performance liquid chromatography (HPLC) in combination with accelerator mass spectrometry (AMS), [26] diode-array detection (DAD), [8] chemical reaction interface for mass spectrometry (CRIMS), [27] UV, [9] as well as capillary electrophoresis (CE) with UV detection. [19][20][21][22][23][24][25] Various other analytical methods have also been applied to detect APAP and its metabolites, such as high performance liquid chromatography (HPLC) in combination with accelerator mass spectrometry (AMS), [26] diode-array detection (DAD), [8] chemical reaction interface for mass spectrometry (CRIMS), [27] UV, [9] as well as capillary electrophoresis (CE) with UV detection.…”

“…[28] In particular, hyphenated HPLC-mass spectrometry (LC-MS) has shown great potential, because of its specificity and its ability to identify structurally-related substances that could bias the result of immunoassay analysis. [25,[34][35][36][37][38][39] An assay for APAP and all major metabolites was reported for rat plasma. [25,[34][35][36][37][38][39] An assay for APAP and all major metabolites was reported for rat plasma.…”

“…Several LC-MS methods have been reported for quantification of APAP [29] or APAP in combination with other analytes, for example other therapeutic drugs, [30][31][32][33] or several metabolites in various matrices. [25] It is essential that methods developed for clinical diagnostics are carefully validated for potential matrix interferences. [25] It is essential that methods developed for clinical diagnostics are carefully validated for potential matrix interferences.…”