Premium
Applying human and pig hepatic in vitro experiments for sulfur mustard study: Screening and identification of metabolites by liquid chromatography/tandem mass spectrometry
Author(s) -
Halme Mia,
Pesonen Maija,
Hakala Ullastiina,
Pasanen Markku,
Vähäkangas Kirsi,
Vanninen Paula
Publication year - 2015
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.7218
Subject(s) - chemistry , sulfur mustard , chromatography , metabolite , mass spectrometry , tandem mass spectrometry , glutathione , electrospray ionization , microsome , liquid chromatography–mass spectrometry , sulfur , biochemistry , in vitro , toxicity , organic chemistry , enzyme
Rationale Sulfur mustard is a chemical warfare agent (CWA) with high toxicity and complex metabolism. This study aimed at identification of new metabolic biomarkers for sulfur mustard using in in vitro exposures and various mass spectrometric techniques. Methods Human and pig liver subcellular fractions were used as biocatalysts. Metabolites were screened by liquid chromatography and tandem mass spectrometry (LC/MS/MS) using positive electrospray ionization (ESI). For structural identification, product ion scans (MS/MS, MS 3 ) and accurate mass measurements using liquid chromatography/time‐of‐flight mass spectrometry (LC/TOFMS) were acquired. Results Sulfur mustard is metabolized in vitro by S‐oxidation and glutathione (GSH) conjugations. One S‐oxidized metabolite, bis(2‐chloroethyl) sulfoxide ( m/z 175), was formed in both species only when liver microsomes were present in incubations, and it was the main metabolite if GSH was not added into the reaction mixture. However, conjugation with GSH was found to be a spontaneous reaction in physiological pH and buffered solution. Three GSH conjugates of sulfur mustard were detected and identified, among which two were novel; 2‐((2‐( S ‐glutathionyl)ethyl)thio)ethanol ( m/z 412) and 2‐((2‐( S ‐glutathionyl)ethyl)thio)ethyl phosphate ( m/z 492). Conclusions To our knowledge, this was the first time that S‐oxidized metabolites and GSH conjugates of sulfur mustard have been detected and identified from human samples in vitro by LC/MS/MS. The usefulness of the GSH conjugates to serve as biomarkers for sulfur mustard exposure in human samples requires further studies. Copyright © 2015 John Wiley & Sons, Ltd.