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Issue 8, 2013
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Quantitative LC/MS/MS analysis of acetaminophen–cysteine adducts (APAP–CYS) and its application in acetaminophen overdose patients

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Abstract

A more sensitive approach by liquid chromatographytandem mass spectrometry (LC/MS/MS) method using the AB Sciex QTRAP 5500 was developed and validated for the detection and quantification of acetaminophencysteine adducts (APAP–CYS) in human plasma. Chromatographic separation was achieved using a Protecol P C18 column (2.1 mm i.d. × 100 mm, 3 microns). The mobile phase consists of A: 2 mM ammonium formate in water (0.2% formic acid) and B: 2 mM ammonium formate in acetonitrile (0.2% formic acid). The analysis was performed using positive and negative ion electrospray ionization (ESI) in multiple reaction monitoring mode (MRM) with a total run time of 7 min per assay. The MS/MS ion transitions monitored were m/z 271 → 140 for APAP–CYS (positive mode) and m/z 154 → 111 for acetaminophen-D4 (negative mode). The newly developed method showed good linearity in the range of 1.0 to 100 ng mL−1. Inter and intra batch precision and accuracy of the method ranged from 0.28 to 5.30% and 87.0 to 113% respectively. LOD was 0.5 ng mL−1 and LLOQ was 1.0 ng mL−1. Analytes were extracted from plasma samples by simple protein precipitation using acetonitrile. The method was successfully applied in a preliminary clinical study to determine APAP–CYS concentration from 46 acetaminophen overdose patients treated at outpatient clinic in the University Malaya Medical Centre.

Graphical abstract: Quantitative LC/MS/MS analysis of acetaminophen–cysteine adducts (APAP–CYS) and its application in acetaminophen overdose patients

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Article information


Submitted
02 Nov 2012
Accepted
05 Feb 2013
First published
07 Feb 2013

Anal. Methods, 2013,5, 1955-1964
Article type
Paper

Quantitative LC/MS/MS analysis of acetaminophencysteine adducts (APAP–CYS) and its application in acetaminophen overdose patients

T. Hairin, A. R. Marzilawati, E. M. H. Didi, S. Mahadeva, Y. K. Lee, N. Abd. Rahman, A. M. Mustafa and Z. Chik, Anal. Methods, 2013, 5, 1955
DOI: 10.1039/C3AY26614A

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