A simultaneous liquid chromatography/mass spectrometric assay of glutathione, cysteine, homocysteine and their disulfides in biological samples

Guan, Xiaoxu; Hoffman, Brianna N.; Dwivedi, Chandradhar; Matthees, Duane P.

doi:10.1016/s0731-7085(02)00594-0

Cited by 275 publications

(178 citation statements)

References 43 publications

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“…The integration windows for each of the scan methods are shown in Table 1, and the LODs of The absolute LODs for the thiols were in the sub-picomole range, and they were better than the LC-UV methods (11,12,33), most of the MS based detection methods (5,(34)(35)(36)(37)(38)(39), and the results via fluorescence measurement (40,41).…”

Section: Figures Of Merit For the Analytical Protocolmentioning

confidence: 99%

Determination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate

Rao

Xiang

Bramanti

et al. 2010

J. Agric. Food Chem.

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A liquid chromatography method with mass spectrometric detection has been developed for the simultaneous determination of six thiols in the sulfur metabolic pathway, including cysteine (Cys), homocysteine (HCys), glutathione (GSH), cysteinyl-glycine (Cys-Gly), γ-glutamyl-cysteine (Glu-Cys), and s-adenosyl-homocysteine (AdoHcy).Tris(2-carboxyethyl) phosphine (TCEP) was used as the reducing reagent and p-hydroxymercurybenzoate (PHMB) as the derivatization reagent. Thiols were extracted from three mg of yeast using water in an ultrasonic bath. The absolute detection limits for the compounds studied were in the sub-picomole range. It was found that AdoHcy, Cys, GSH, Cys-Gly, Glu-Cys, and very little HCys were present in the selenium enriched yeast sample studied, and GSH, Glu-Cys, very little AdoHcy, Cys, and Cys-Gly were present in three bakery yeasts.

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Section: Figures Of Merit For the Analytical Protocolmentioning

confidence: 99%

Determination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate

Rao

Xiang

Bramanti

et al. 2010

J. Agric. Food Chem.

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“…The finding of no Hb adducts induced by BFBDS or p-FBM indicated the formation of Hb dmd.aspetjournals.org adducts was based on thiol/disulfide exchange, and the structure of trisulfide meant higher reactivity toward thiols. The reason why BFBTS-derived adducts were not detected at low concentration was probably due to the great abundance of GSH in RBC (;2 mM) of rats (Guan et al, 2003). Our previous data revealed the rapid metabolism of BFBTS by GSH even at low concentration.…”

Section: Discussionmentioning

confidence: 86%

Metabolism of Bis(4-fluorobenzyl)trisulfide and Its Formation of Hemoglobin Adduct in Rat Erythrocytes

Pān

Sun

et al. 2013

Drug Metab Dispos

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Bis(4-fluorobenzyl)trisulfide (BFBTS) is a promising new antitumor agent under investigation. It was metabolized rapidly in vivo in rat, but the metabolic fate and primary site of metabolism have not been clarified. In this study, we investigated the role of blood in the metabolism of BFBTS and compared the BFBTS metabolic potencies in whole blood, plasma, and red blood cells (RBCs) in vitro. Three major metabolites of BFBTS [bis(4-fluorobenzyl) disulfide, para-fluorobenzyl-mercaptan, and para-fluorobenzoic acid] were detected in RBCs and whole blood. Significant metabolism of BFBTS was observed in RBCs that were identified as the primary site of BFBTS metabolism. Thiols, including endogenous thiols and hemoglobin, were proven to be the critical factor in BFBTS metabolism. S-Fluorobenzylmercaptocysteine Hb (hemoglobin) adducts were characterized in vitro at BFBTS concentration of 250 mM and higher, whereas such Hb adducts were not detected in RBCs from Sprague-Dawley rats receiving a single intravenous injection of BFBTS at a high dose of 50 mg/kg. Liquid chromatography-tandem mass spectrometry results revealed that adduction induced by BFBTS was prone to take place at Cys125 of globin b chains. Otherwise, glutathionylation of Hb was also observed that may be attributed to the oxidative effect of BFBTS. In summary, BFBTS was unstable when it met with thiols, and RBCs were the main site of BFBTS metabolism. Hb adducts induced by BFBTS could be detected in vitro at high concentration but not in vivo even at high dose.

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“…This was the spectrophotometric method with Ellman's reagent (5,5-dithio-bis-2-nitrobenzoic acid). According to Guan et al (2003), the addition of Ellman's reagent increases the analyte chemical stability and accuracy of the test, and protects the protein thiols (Guan et al 2003). In the fluorometric methods, the authors applied o-phthaldialdehyde (OPA) as the fluorescent reagent to a reaction with amino acids which was used to determine GSH and GSSG (Cohn and Lyle 1966;Hissin and Hilf 1976).…”

Section: Short Review Of a Few Techniques Often Used To Determine Glumentioning

confidence: 99%

The Use of Different MS Techniques to Determine Glutathione Levels in Marine Tissues

2012

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Several techniques were used, mainly mass spectrometry connected with gas chromatography, matrixassisted laser desorption with ionization time-of-flight and mass spectrometry connected with liquid chromatography. A major problem was encountered in determining glutathione, which can be conditioned by the pH and selected reducing agents. The GSH form can be oxidized through derivatization, and a small glutathione amount in the biological samples may hinder the determination process. Another problem is the existence of a metal ion in the tested organism; therefore, often a reagent with a chelating function is added to the sample and the mobile phase in liquid chromatography is applied with appropriate polarity for GSH and GSSG. We determined the concentrations of total, reduced, and oxidized glutathione in the liver, hepatopancreas, muscle, and gonad tissues of brown shrimp (Crangon crangon) and fish (Psetta maxima and Clupea harengus membras). The highest concentrations of tGSH were recorded in the shrimp hepatopancreas (7.21 ± 0.011 μmol g −1 wet weight), in herring liver (2.85±0.025 μmol g −1 ), and in turbot liver (1.86±0.063 μmol g −1). In turn, the highest concentrations were reported for GSSG in the muscle of shrimp (0.140±0.000204 μmol g ). We also investigated seasonal changes in the concentrations of glutathione in the muscle of C. crangon shrimp in the annual cycle. The lowest values of total glutathione were recorded during spring and autumn, which could be correlated with the increase in lipid peroxidation and oxidative stress.

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A simultaneous liquid chromatography/mass spectrometric assay of glutathione, cysteine, homocysteine and their disulfides in biological samples

Cited by 275 publications

References 43 publications

Determination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate

Determination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate

Metabolism of Bis(4-fluorobenzyl)trisulfide and Its Formation of Hemoglobin Adduct in Rat Erythrocytes

The Use of Different MS Techniques to Determine Glutathione Levels in Marine Tissues

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