Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 3, March 2008, pp. 209-214
Evaluation of BEH C18, BEH HILIC, and HSS T3
(C18) Column Chemistries for the UPLC–MS–MS Analysis
of Glutathione, Glutathione Disulfide, and Ophthalmic Acid in
Mouse Liver and Human Plasma
Lee-Sun New and Eric C.Y. Chan
Department of Pharmacy, Faculty of Science, National University
of Singapore, 18 Science Drive 4, Singapore 117543
Glutathione (GSH), glutathione disulfide (GSSG),
and ophthalmic acid (OA) are important biological oxidative stress
biomarkers to be monitored in pathological and toxicological
studies. With the advent of liquid chromatography tandem mass
spectrometry (LC–MS–MS) technology, sensitive and
selective analysis of these biomarkers in biological samples
is now being performed routinely. Due to the hydrophilic and
polar natures of GSH and its endogenous derivatives, achieving
good retention, resolution, and peak shape is often a chromatographic
challenge. In this study, three ultra-performance (UP) LC column
chemistries (namely, BEH C18, BEH HILIC, and HSS T3 [C18]) are
evaluated for the UPLC–MS–MS analysis of GSH, GSSG,
and OA extracted from mouse liver and human plasma samples. The
chromatographic parameters evaluated are retentivity, tailing
factor, MS sensitivity, and resolution of the three analytes.
Based on the optimized method for each column chemistry, our
results indicate that the HSS T3 (C18) column chemistry affords
the best retention and separation of these analytes when operated
under the ultra high-pressure chromatographic conditions.
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