Sensitive determination of glutathione in biological samples by capillary electrophoresis with green (515 nm) laser-induced fluorescence detection

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Authors

HODÁKOVÁ Júlia PREISLER Jan FORET František KUBÁŇ Petr

Year of publication 2015
Type Article in Periodical
Magazine / Source Journal of Chromatography A
MU Faculty or unit

Faculty of Science

Citation
Doi http://dx.doi.org/10.1016/j.chroma.2015.02.062
Field Analytic chemistry
Keywords Capillary electrophoresis; Laser-induced fluorescence; Glutathione; Eosin-5-maleimide
Description A new sensitive capillary electrophoretic method with laser-induced fluorescence (LIF) was developed for quantitation of glutathione (GSH) in biological samples. Eosin-5-maleimide was used to label the GSH molecule and the formed conjugate was separated in a 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid electrolyte at pH 7.0 in less than 3 min. The conjugate was detected with an in-house built LIF system, utilizing an inexpensive 515 nm diode laser module. Studies were performed to optimize the derivatizatiory(the ratio of reagent to analyte, the reaction time, pH, etc.) and separation conditions. Sensitive detection of GSH at concentrations as low as 0.18 nM was obtained. The method was applied in the analysis of biological fluids (exhaled breath condensate, saliva) and was found to be suitable for determination of GSH in these samples at trace levels below 1 nM. To the best of our knowledge, this is the first report on determination of GSH in exhaled breath condensate by capillary electrophoresis (CE). (C) 2015 Elsevier B.V. All rights reserved.
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