Quantitation of oxidized and reduced glutathione in plasma by micellar electrokinetic capillary electrophoresis

Havel, Karalyn; Pritts, Kathy; Wielgos, Todd

doi:10.1016/s0021-9673(99)00518-x

Cited by 42 publications

(27 citation statements)

References 17 publications

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“…There have been two reports on the determination of water samples (waste water or river water), 14 alkylphenols (4-substituted, n:1-2 or halogene) in water by capillary zone electrophoresis (CZE; Gaitonde and Pathak, 1990), bisphenol A and 3 alkylphenols in water by micellar electrokinetic chromatography (MEKC; Takeda et al, 2000). In particular, MEKC was suitable for analysis of low molecular weight compounds and drugs (Takeda et al, 1993(Takeda et al, , 2000Terabe et al, 1984;Chan et al, 1993;Havel et al, 1999;Kobayashi et al, 1995;Imazawa and Hatanake, 1997). MEKC has high separation efficiency and uses small volumes of migration buffer without organic solvents.…”

Section: Introductionmentioning

confidence: 99%

Determination of bisphenol A and 10 alkylphenols in serum using SDS micelle capillary electrophoresis with γ‐cyclodextrin

Katayama

Matsuda

Sasaki

et al. 2001

Biomedical Chromatography

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A new micelle capillary electrophoresis based on cyclodextrin micellar electrokinetic chromatography (MEKC) for the determination of bisphenol A and 10 alkylphenols in rat serum is reported. Several surfactants and dextrins were studied. Bisphenol A and alkylphenols were separated using a 50 microm x 50 cm capillary with 20 mM borate phosphate buffer (pH 8.0) containing 20 mM sodium dodecylsulfate and 5 mM gamma-cyclodextrin as carrier. The method could determine 0.6-2000 microg/mL of phenols in 100 microL serum by photometric detection at 214 nm. Using 2.0 mL serum, 1.0 ng/mL of phenols could be determined. The relative standards deviations were 6.3-7.7% at 10 microg/mL in serum. The recoveries were 91.8-93.0% with 10 microg/mL serum samples.

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Section: Introductionmentioning

confidence: 99%

Determination of bisphenol A and 10 alkylphenols in serum using SDS micelle capillary electrophoresis with γ‐cyclodextrin

Katayama

Matsuda

Sasaki

et al. 2001

Biomedical Chromatography

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“…The ratio of GSH to GSSG was 17.8 6 1.3 for heart tissue, whereas it was much higher (.100) in the mitochondria [59]. Quite the same strategy of analysis was used by Havel et al [60]. Figure 6 presents the results obtained on plasma.…”

Section: Oxidized and Reduced Forms Of Gshmentioning

confidence: 88%

Determination of aminothiols in body fluids, cells, and tissues by capillary electrophoresis

2004

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Oxidative stress is present in cardiovascular diseases and hyperhomocysteinemia, an independent risk factor for these diseases. It may play a role by inducing production of oxygen free radicals. Reduced glutathione is the most abundant intracellular low-molecular-weight thiol and plays an essential role in protecting cells from toxic species. The thiol-containing compounds which are the most often considered in biological analysis, are homocysteine (Hcy), cysteine (Cys), glutathione (GSH), cysteinyl-glycine (Cys-Gly), gamma-glutamyl-cysteine (gammaGlu-Cys), and their derivatives. These aminothiols are present in body fluids or cells, associated with proteins or occur free (reduced and oxidized). These free forms may play a role in the pathogenesis of disease. Because Hcy (with Cys) exhibits pro-oxidative properties and GSH (with Cys-Gly) antioxidative properties, and because there is extensive interconversion between these metabolites, their simultaneous analysis in biological samples is necessary to examine their role in human disease. Capillary electrophoresis (CE) seems to be a solution to reach this goal. No extensive review reports the analysis of aminothiols using CE. This review describes the different CE approaches which have been used to separate and assay aminothiols, and the different obtained datas.

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“…Although liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the technique of choice for the simultaneous determination of GSH and GSSG (taking into consideration specificity and low limits of detection and quantification), it is inaccessible to many laboratories [11][12][13]. Other methods, such as micellar electrokinetic capillary electrophoresis (MEKC) [14], nuclear magnetic resonance (NMR) [15], fluorescence [16], and chemiluminescence-based methods, have also been proposed [17]. The lack of total automatization and inability to determine protein-bound glutathione are the major limitations of capillary electrophoresis; NMR is not commonly available in clinical laboratories; fluorescence requires previous derivatization, while GSSG determination is not achievable by chemiluminescence methods.…”

Section: Introductionmentioning

confidence: 99%

The simple isocratic HPLC—UV method for the simultaneous determination of reduced and oxidized glutathione in animal tissue

Begić¹,

Djurić²,

Gobeljić³

et al. 2017

Acta Chromatographica

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Summary. The aim of our work was to optimize and apply simple high-performance liquid chromatography method with ultraviolet detection (HPLC-UV) for simultaneous determination of reduced (GSH) and oxidized (GSSG) glutathione in biological matrix (specifically, the rat liver tissue was used herein), since the ratio between oxidized and reduced glutathione forms (GSSG-GSH) has been recognized as an important biological marker of oxidatively depleted GSH in oxidative stress (OS)-associated diseases and poisonings. An isocratic chromatographic separation of GSH and GSSG (2.8 min and 6.3 min, respectively) was performed with the mobile phase consisted of sodium perchlorate solution (pH adjusted to 2.8) at flow rate of 1 mL min −1 , detection set at 215 nm, and column temperature of 40 °C. The method offers short run time, linearity in the range of 0.01-200 µM concentration for both compounds (R 2 = 1), low limits of detection and quantification (GSH: 0.18 µM and 0.56 µM, GSSG: 0.52 µM and 1.58 µM, respectively), precision, accuracy (bias < 2%), and high reproducibility.Through suitable sample handling, an overestimation of GSSG was prevented. High recovery (>99%) was achieved. The method was successfully applied for the analysis of GSH and GSSG in liver homogenates of Wistar rats intraperitoneally exposed to cadmium (Cd) (1 mg kg −1 CdCl2/21 days). Regardless of other Cd-mediated hepatotoxicity mechanisms, herein, we have exclusively interpreted/emphasized oxidative GSH depletion.The presented method is acceptable for a routine analysis of GSH and GSSG in biological matrix, while the calculated ratio GSSG-GSH is considered as a valuable OS marker.

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Quantitation of oxidized and reduced glutathione in plasma by micellar electrokinetic capillary electrophoresis

Cited by 42 publications

References 17 publications

Determination of bisphenol A and 10 alkylphenols in serum using SDS micelle capillary electrophoresis with γ‐cyclodextrin

Determination of bisphenol A and 10 alkylphenols in serum using SDS micelle capillary electrophoresis with γ‐cyclodextrin

Determination of aminothiols in body fluids, cells, and tissues by capillary electrophoresis

The simple isocratic HPLC—UV method for the simultaneous determination of reduced and oxidized glutathione in animal tissue

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