2020
DOI: 10.3390/molecules25071735
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Analysis of Total Thiols in the Urine of a Cystathionine β-Synthase-Deficient Mouse Model of Homocystinuria Using Hydrophilic Interaction Chromatography

Abstract: Homocysteine and related thiols (cysteine, cysteinylglycine, and glutathione) in the urine of a cystathionine β-synthase (CBS)-deficient mouse model were quantified using hydrophilic interaction chromatography with fluorescence detection. Urine samples were incubated with tris(2-carboxyethyl) phosphine to reduce disulfide bonds into thiols. After deproteinization, thiols were fluorescently derivatized with ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate (SBD-F). Homocysteine, cysteine, cysteinylglycine, and… Show more

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Cited by 4 publications
(4 citation statements)
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“…Good linearity was obtained, with correlation coefficients exceeding 0.999. The thiol sensitivity was lower than that obtained with HILIC methods [14][15][16][17] and was similar to those obtained in other studies [23,24], except for GSH. This was because the sensitivity of the fluorescence detector for GSH was lowered.…”
Section: Methods Validationsupporting
confidence: 88%
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“…Good linearity was obtained, with correlation coefficients exceeding 0.999. The thiol sensitivity was lower than that obtained with HILIC methods [14][15][16][17] and was similar to those obtained in other studies [23,24], except for GSH. This was because the sensitivity of the fluorescence detector for GSH was lowered.…”
Section: Methods Validationsupporting
confidence: 88%
“…In this study, we developed an analytical method for detecting and quantifying intracellular thiols that can be applied to K562 cell samples. Thiols were first reduced by TCEP and then fluorescently derivatized by SBD-F. Six different SBD-thiols were successfully separated within 30 min on an ODS column, which can perform more stable analyses than HILIC columns [14][15][16][17]. By adjusting the sensitivity of the fluorescence detector, Cys, CysGly, γGC, and GSH in K562 cells could be quantified simultaneously.…”
Section: Discussionmentioning
confidence: 99%
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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%