Ultra-high performance hydrophilic interaction liquid chromatography – Triple quadrupole tandem mass spectrometry method for determination of cysteine, homocysteine, cysteinyl-glycine and glutathione in rat plasma

Forgacsova, Andrea; Galba, Jaroslav; Mojžišová, Jana; Mikuš, Peter; Piešťanský, Juraj; Kováč, Andrej

doi:10.1016/j.jpba.2018.10.053

Cited by 55 publications

(18 citation statements)

References 31 publications

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“…The assays based on gas/liquid chromatography–mass spectrometric detection [ 11 , 12 , 13 ] or glutathione reductase-assisted optical detection [ 14 , 15 ] are established ones for the analysis of both GSH and GSSG. The above techniques, however, require cell lysis accompanied by difficult-to-control autooxidation of the GSH pool, hardly consider rapid redox fluctuations and are also laborious and/or sufficiently expensive for high-throughput tasks.…”

Section: Introductionmentioning

confidence: 99%

Probing Cell Redox State and Glutathione-Modulating Factors Using a Monochlorobimane-Based Microplate Assay

et al. 2022

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Thiol compounds including predominantly glutathione (GSH) are key components of redox homeostasis, which are involved in the protection and regulation of mammalian cells. The assessment of cell redox status by means of in situ analysis of GSH in living cells is often preferable over established assays in cell lysates due to fluctuations of the GSH pool. For this purpose, we propose a microplate assay with monochlorobimane (MCB) as an available fluorescent probe for GSH, although poorly detected in the microplate format. In addition to the new procedure for improved MCB-assisted GSH detection in plate-grown cells and its verification with GSH modulators, this study provides a useful methodology for the evaluation of cell redox status probed through relative GSH content and responsiveness to both supplemented thiols and variation in oxygen pressure. The roles of extracellular interactions of thiols and natural variability of cellular glutathione on the assay performance were emphasized and discussed. The results are of broad interest in cell biology research and should be particularly useful for the characterization of pathological cells with decreased GSH status and increased oxidative status as well as redox-modulating factors.

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Section: Introductionmentioning

confidence: 99%

Probing Cell Redox State and Glutathione-Modulating Factors Using a Monochlorobimane-Based Microplate Assay

et al. 2022

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“…There are several procedures based on LC–MS/MS for measuring Hcy with Cys and Met (Bártl et al, 2014; Forgacsova et al, 2019; Jiang, Mistretta, Elsea, & Sun, 2017). The most obvious disadvantage is the chromatographic speed.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous determination of serum homocysteine, cysteine, and methionine in patients with schizophrenia by liquid chromatography–tandem mass spectrometry

Liu²,

Chen³

et al. 2022

Biomedical Chromatography

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Schizophrenia is a debilitating psychiatric disorder affecting approximately 1% of the population worldwide. Disturbances in the homocysteine metabolism are an important factor in the pathophysiology of schizophrenia. In this research, a novel validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantification procedure was developed to investigate three significant compounds of homocysteine metabolism: homocysteine, cysteine, and methionine in patients with schizophrenia and healthy controls. Sample preparation involved a reduction with dithiothreitol followed by protein precipitation, and the chromatographic runtime was 2 min. The LC-MS/MS method was validated according to CLSI C62-A and the Chinese Guidance for Liquid Chromatography and Mass Spectrometry Clinical Application. The performance of the method was excellent with a coefficient of variation for precision in the range of 0.5-6.9%, an accuracy of 90.4-101.6%. In addition, the practical applicability of the method was demonstrated by applying it in the routine sample analysis for a schizophrenic patient. Increased homocysteine levels and decreased cysteine levels were observed in the patient with schizophrenia. These results indicate that the activity of the transsulfuration pathway may play a key role in the pathogenesis of schizophrenia.

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“…So far, a great number of methods have been reported for detecting Hcy, including radioenzyme immunoassay (RIA) [7], high performance liquid chromatography (HPLC) [8][9][10][11][12][13], electrochemistry [2,14], capillary electrophoresis (CE) [15,16], colorimetry [1,[17][18][19][20], liquid chromatography-mass spectrometry (LC-MS) [21][22][23], etc. [24].…”

Section: Introductionmentioning

confidence: 99%

Simple and fast detection of homocysteine by cucurbit[7]uril fluorescent probe based on competitive strategy

Jiang

Xiao

Yang

et al. 2022

Chemical Physics Letters

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This research put forward a simple and rapid fluorescence probe method based on competition strategy to detect homocysteine. The host-guest interaction behavior between cucurbit[7]uril and homocysteine was further studied via UV-vis spectroscopy, Fourier transform infrared spectroscopy, molecular modeling, and 1 H NMR spectroscopy in aqueous solution. The binding constant of cucurbit [7]uril-homocysteine measured by 1 H NMR titration is 2.40  10 3 L mol -1 .Under the optimal detection conditions, the fluorescence intensity of the cucurbit[7]uril-berberine complex declined linearly with increasing concentration of homocysteine in the range from 5 to 200 µM, and the limit of detection was 1.76 µM. This established fluorescent probe method exhibits high selectivity for coexisting interfering substances. Meanwhile, it was also successfully employed to detect homocysteine in serum samples with the recovery rate of 85.1% to 102.0%. This supramolecular fluorescent probe based on competitive strategy provides a simple and fast method for the determination of homocysteine in practical samples.

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Ultra-high performance hydrophilic interaction liquid chromatography – Triple quadrupole tandem mass spectrometry method for determination of cysteine, homocysteine, cysteinyl-glycine and glutathione in rat plasma

Cited by 55 publications

References 31 publications

Probing Cell Redox State and Glutathione-Modulating Factors Using a Monochlorobimane-Based Microplate Assay

Probing Cell Redox State and Glutathione-Modulating Factors Using a Monochlorobimane-Based Microplate Assay

Simultaneous determination of serum homocysteine, cysteine, and methionine in patients with schizophrenia by liquid chromatography–tandem mass spectrometry

Simple and fast detection of homocysteine by cucurbit[7]uril fluorescent probe based on competitive strategy

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