Determination of biologically active low-molecular-mass thiols in human blood

Simultaneous determination of total homocysteine, cysteine, cysteinylglycine, and glutathione in human plasma by high-performance liquid chromatography: Application to studies of oxidative stress

“…This in turn has generated a need for fully validated, simple, precise and sensitive analytical methods capable of simultaneously determining aminothiols for clinical and translational research and for routine clinical monitoring. Numerous methods for the determination of one or more thiols have been reported, including HPLC with ultraviolet [7][8][9], fluorescence [10][11][12][13][14][15][16][17][18]31], or electrochemical detection [19][20][21], gas chromatography-mass spectrometry [22], ionexchange chromatography [23], and capillary electrophoresis [24]. Each of these methods has basic limitations in terms of equipment cost, complexity, sample processing and run times, number of thiols simultaneously quantitated, and/or validation parameters assessed which hinder their use for high-throughput routine clinical or research purposes.…”

Simultaneous analysis of multiple aminothiols in human plasma by high performance liquid chromatography with fluorescence detection

“…Fluorogenic reagents used to detect aminothiols in plasma include monobromobimane (mBrB) [32][33][34][35][36]39], ophthaldialdehyde (OPA) [27,28], dansyl chloride [29,30], and ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F) [15,26,31,37,38,40]. Monobromobimane reacts rapidly, but not specifically, with thiols at pH 8.0 at room temperature.…”

“…Assays capable of determining multiple analytes are desirable because of extensive interconversion of aminothiols and the need to understand the dynamic relationship between all thiol and disulfide components [16,17], as well as to facilitate assessment of the relationship between thiols and disease status. A variety of methods are available for determining concentrations of two or more aminothiols, including capillary electrophoresis [18,19], gas or liquid chromatography with mass spectrometric detection [20][21][22], and HPLC with ultraviolet [23][24][25], fluorescence [15,[26][27][28][29][30][31][32][33][34][35][36][37][38][39][40], or electrochemical detection [41][42][43][44]. Each of these methods has basic limitations in terms of performance, equipment cost, complexity, sample processing and run times, and/or validation parameters assessed, which create challenges or render them impractical for high-throughput routine clinical or research purposes.…”

Reaction‐Based Europium Complex for Specific Detection of Cysteine Over Homocysteine and Glutathione with Variable‐Temperature Kinetic Studies

“…Classical analytical methods for detection of Cys such as high‐performance liquid chromatography (HPLC), potentiometry, and capillary electrophoresis have some drawbacks, including complicated and laborious pretreatment procedures and expensive instrumentation. Compared with other techniques, fluorimetry is an excellent and simple detection method due to its sensitivity, selectivity, convenience, and low cost.…”