Robert B Flint1, Paola Mian, Bart van der Nagel, Nuria Slijkhuis, Birgit C P Koch. 1. *Department of Pharmacy, Erasmus University Medical Center; †Department of Pediatrics, Division of Neonatology, Erasmus University Medical Center-Sophia, Rotterdam; ‡Department of Pharmacy, Radboudumc, Nijmegen; and §Intensive Care and Department of Pediatric Surgery, Erasmus University Medical Center-Sophia, Rotterdam, the Netherlands.
Abstract
BACKGROUND: Acetaminophen (APAP, paracetamol) is the most commonly used drug for pain and fever in both the United States and Europe and is considered safe when used at registered dosages. Nevertheless, differences between specific populations lead to remarkable changes in exposure to potentially toxic metabolites. Furthermore, extended knowledge is required on metabolite formation after intoxication, to optimize antidote treatment. Therefore, the authors aimed to develop and validate a quick and easy analytical method for simultaneous quantification of APAP, APAP-glucuronide, APAP-sulfate, APAP-cysteine, APAP-glutathione, APAP-mercapturate, and protein-derived APAP-cysteine in human plasma by ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry. METHODS: The internal standard was APAP-D4 for all analytes. Chromatographic separation was achieved with a reversed-phase Acquity ultraperformance liquid chromatography HSS T3 column with a runtime of only 4.5 minutes per injected sample. Gradient elution was performed with a mobile phase consisting of ammonium acetate, formic acid in Milli-Q ultrapure water or in methanol at flow rate of 0.4 mL/minute. RESULTS: A plasma volume of only 10 μL was required to achieve both adequate accuracy and precision. Calibration curves of all 6 analytes were linear. All analytes were stable for at least 48 hours in the autosampler; the high quality control of APAP-glutathione was stable for 24 hours. The method was validated according to the U.S. Food and Drug Administration guidelines. CONCLUSIONS: This method allows quantification of APAP and 6 metabolites, which serves purposes for research, as well as therapeutic drug monitoring. The advantage of this method is the combination of minimal injection volume, a short runtime, an easy sample preparation method, and the ability to quantify APAP and all 6 metabolites.
BACKGROUND:Acetaminophen (APAP, paracetamol) is the most commonly used drug for pain and fever in both the United States and Europe and is considered safe when used at registered dosages. Nevertheless, differences between specific populations lead to remarkable changes in exposure to potentially toxic metabolites. Furthermore, extended knowledge is required on metabolite formation after intoxication, to optimize antidote treatment. Therefore, the authors aimed to develop and validate a quick and easy analytical method for simultaneous quantification of APAP, APAP-glucuronide, APAP-sulfate, APAP-cysteine, APAP-glutathione, APAP-mercapturate, and protein-derived APAP-cysteine in human plasma by ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry. METHODS: The internal standard was APAP-D4 for all analytes. Chromatographic separation was achieved with a reversed-phase Acquity ultraperformance liquid chromatography HSS T3 column with a runtime of only 4.5 minutes per injected sample. Gradient elution was performed with a mobile phase consisting of ammonium acetate, formic acid in Milli-Q ultrapure water or in methanol at flow rate of 0.4 mL/minute. RESULTS: A plasma volume of only 10 μL was required to achieve both adequate accuracy and precision. Calibration curves of all 6 analytes were linear. All analytes were stable for at least 48 hours in the autosampler; the high quality control of APAP-glutathione was stable for 24 hours. The method was validated according to the U.S. Food and Drug Administration guidelines. CONCLUSIONS: This method allows quantification of APAP and 6 metabolites, which serves purposes for research, as well as therapeutic drug monitoring. The advantage of this method is the combination of minimal injection volume, a short runtime, an easy sample preparation method, and the ability to quantify APAP and all 6 metabolites.
Authors: P Mian; M J van Esdonk; K T Olkkola; B C M de Winter; A Liukas; I Spriet; D Tibboel; M Petrovic; B C P Koch; K Allegaert Journal: Br J Clin Pharmacol Date: 2018-10-25 Impact factor: 4.335
Authors: L T van der Heijden; P Mian; J Hias; B C M de Winter; J Tournoy; L Van der Linden; D Tibboel; K Walgraeve; J Flamaing; B C P Koch; K Allegaert; I Spriet Journal: Drugs Aging Date: 2021-12-17 Impact factor: 3.923
Authors: Paola Mian; Karel Allegaert; Sigrid Conings; Pieter Annaert; Dick Tibboel; Marc Pfister; Kristel van Calsteren; John N van den Anker; André Dallmann Journal: Clin Pharmacokinet Date: 2020-07 Impact factor: 6.447
Authors: P Mian; A J Valkenburg; K Allegaert; B C P Koch; C V Breatnach; C A J Knibbe; D Tibboel; E H J Krekels Journal: J Clin Pharmacol Date: 2019-01-11 Impact factor: 3.126
Authors: Paola Mian; John N van den Anker; Kristel van Calsteren; Pieter Annaert; Dick Tibboel; Marc Pfister; Karel Allegaert; André Dallmann Journal: Clin Pharmacokinet Date: 2020-01 Impact factor: 6.447
Authors: L M Koehn; Y Huang; M D Habgood; S Nie; S Y Chiou; R B Banati; K M Dziegielewska; N R Saunders Journal: Sci Rep Date: 2021-10-06 Impact factor: 4.379