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Absolute quantification of free glutathione and cysteine in aquatic insects using isotope dilution and selected reaction monitoring

Absolute quantification of free glutathione and cysteine in aquatic insects using isotope... A simple and robust isotope dilution mass spectrometry-based assay was developed for the determination of free cysteine and glutathione (GSH) in aquatic insects. Several experimental parameters were evaluated and optimized to provide specific and sensitive detection of both compounds by in situ derivatization with N-ethylmaleimide followed by acid alkylation quenching and reverse-phased liquid chromatography coupled with selected reaction monitoring. For both targets, the assay was evaluated over a concentration range of 0.313 to 320 μM and was demonstrated to have a quantitative dynamic range spanning nearly three orders of magnitude, with lower limits of quantification being 0.330 μM for GSH and 0.370 μM for cysteine. Additionally, measurements were observed to be highly reproducible over the course of several days. When applied to the analysis of four different species of insects, large biological variation between and within species was observed. Different feeding regimens were also tested within two species of insects but statistical comparisons revealed no significant difference in the levels of either compound. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Analytical and Bioanalytical Chemistry Springer Journals

Absolute quantification of free glutathione and cysteine in aquatic insects using isotope dilution and selected reaction monitoring

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References (45)

Publisher
Springer Journals
Copyright
Copyright © 2011 by Springer-Verlag
Subject
Chemistry; Food Science; Laboratory Medicine; Biochemistry, general; Characterization and Evaluation of Materials; Analytical Chemistry; Environmental Monitoring/Analysis
ISSN
1618-2642
eISSN
1618-2650
DOI
10.1007/s00216-011-5416-2
pmid
21956263
Publisher site
See Article on Publisher Site

Abstract

A simple and robust isotope dilution mass spectrometry-based assay was developed for the determination of free cysteine and glutathione (GSH) in aquatic insects. Several experimental parameters were evaluated and optimized to provide specific and sensitive detection of both compounds by in situ derivatization with N-ethylmaleimide followed by acid alkylation quenching and reverse-phased liquid chromatography coupled with selected reaction monitoring. For both targets, the assay was evaluated over a concentration range of 0.313 to 320 μM and was demonstrated to have a quantitative dynamic range spanning nearly three orders of magnitude, with lower limits of quantification being 0.330 μM for GSH and 0.370 μM for cysteine. Additionally, measurements were observed to be highly reproducible over the course of several days. When applied to the analysis of four different species of insects, large biological variation between and within species was observed. Different feeding regimens were also tested within two species of insects but statistical comparisons revealed no significant difference in the levels of either compound.

Journal

Analytical and Bioanalytical Chemistry – Springer Journals

Published: Sep 29, 2011

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