Direct Determination of Tissue Aminothiol, Disulfide, and Thioether Levels Using HPLC-ECD with a Novel Stable Boron-Doped Diamond Working Electrode

Abstract

This chapter describes two different methods using reversed-phase HPLC with electrochemical detection on a boron-doped diamond (BDD) working electrode for the direct, routine, sensitive and simultaneous measurement of a number of aminothiols, disulfides, and thioethers, in either plasma or tissue homogenates.

Evidence of hydrogen sulfide involvement in amyotrophic lateral sclerosis

“…Briefly, the samples (CSF, sera, and tissues) were prepared according to a standard procedure described by Bailey et al with modifications. The HPLC system used to measure H 2 S consisted of an autosampler, an isocratic pump (Dionex ICS 3000; Thermo Fisher Scientific, Waltham, MA), and an amperometric detector.…”

Methodology for a rapid and simultaneous determination of total cysteine, homocysteine, cysteinylglycine and glutathione in plasma by isocratic RP-HPLC

“…These aminothiols ( Fig. 1) are involved in cellular protection against oxygen and nitrogen reactive species, in heavy metal and xenobiotics detoxification, in control of gene expression and in cell signaling [4][5][6]. In fact, thiol groups ( SH) are critical intra and extracellular redox buffers which promptly undergo oxidative coupling reactions to form disulfides ( S S ).…”

“…Over the years, several analytical methods have been developed for thiols determination such as liquid chromatography (LC) [20,21], gas chromatography [22,23], ion-exchange chromatography [24,25] and capillary electrophoresis [26,27]. High performance liquid chromatography (HPLC) with several detection techniques, such as ultraviolet [4,28,29], fluorescence (FL) [30][31][32], electrochemical [5,6,33] and mass spectrometry [34][35][36], is the most reported methodology. All the referred methods have basic limitations in terms of equipment, reagent costs, complexity, sample preparation, run time, number of thiols simultaneously quantified, and/or validation assessment, which delay their use for high-throughput routine clinical or research purposes [8].…”

Increasing tetrahydrobiopterin in cardiomyocytes adversely affects cardiac redox state and mitochondrial function independently of changes in NO production

“…Supernatants obtained from centrifugation at 15,000 × g for 45 min of acid precipitation were loaded onto a Synergi-Hydro column (25-×4.6 mm, Phenomenex, Torrance CA) equilibrated with 25 mM phosphate buffer containing 1.4 mM 1-octanesulfonic acid, 6% v/v acetonitrile, pH 2.6 and eluted at a flow of 0.5 ml/min. The system consisted of an ESA automated injection system, an ESA 582 pump, and amperometric cell (5040) with a BDD working electrode operating at an applied voltage of 1500 mV [29]. GSH and GSSG standards were used for quantification and concentrations were normalized to protein content.…”