HPLC Determination of Bioactive Sulfur Compounds, Amino Acids and Biogenic Amines in Biological Specimens

Abstract

There is an increasing interest for analytical methods aimed to detect biological sulfur-containing amines, because of their involvement in human diseases and metabolic disorders. This work describes an improved HPLC method for the determination of sulfur containing amino acids and amines from different biological matrices. We optimized a pre-column derivatization procedure using dabsyl chloride, in which dabsylated products can be monitored spectrophotometrically at 460 nm. This method allows the simultaneous analysis of biogenic amines, amino acids and sulfo-amino compounds including carnosine, dopamine, epinephrine, glutathione, cysteine, taurine, lanthionine, and cystathionine in brain specimens, urines, plasma, and cell lysates. Moreover, the method is suitable for the study of physiological and non-physiological derivatives of taurine and glutathione such as hypotaurine, homotaurine, homocysteic acid and S-acetylglutathione. The present method displays good efficiency of derivatization, having the advantage to give rise to stable products compared to other derivatizing agents such as o-phthalaldehyde and dansyl chloride.With this method, we provide a tool to study sulfur cycle from a metabolic point of view in relation to the pattern of biological amino-compounds, allowing researchers to get a complete scenario of organic sulfur and amino metabolism in tissues and cells.

Proline synthesis in developing microspores is required for pollen development and fertility

“…The extract was centrifuged at 14,000 g for 20 min and the supernatant reduced to 7 μl under vacuum. The amino acids present in the extract were derivatized with DABS (4-N, N-dimethylaminoazobenzene-4′-sulfonyl chloride, Sigma-Aldrich, USA), according to Francioso et al (2017) [55]. Gradient grade solvents used for chromatographic analyses were purchased from Carlo Erba Reagents (Milan, Italy).…”

Formation, Analytical Methods, Change Tendency, and Control Strategies of Biogenic Amines in Canned Aquatic Products: A Systematic Review

“…BAs are usually extracted by acids such as trichloroacetic acid (TCA), perchloric acid (PCA) or organic solvents such as acetonitrile and methanol. The extraction of biogenic amines is usually performed using TCA, which is preferred over methanol and PCA due to the specificity of methanol for aromatic amines and the risk of damages by handling the PCA, as it is explosive (29). The separation and quantification of BAs are affected by the several compounds present in canned aquatic products such as proteins, fats, free amino acids, polyacids, sugars and other components, which also reduce the detection sensitivity (26).…”

Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

“…In human blood, glutathione levels are much higher in erythrocytes than in plasma and are often difficult to measure because of the presence of ferric ions (Fe 3+ ), which accelerate the oxidation of GSH to GSSG (disulfide oxidized glutathione) [2]. There are several methodologies, mostly HPLC-based, for the determination of GSH in different biological samples (GSH, GSSG, total GSH) [3][4][5][6][7]. The most common methods for HPLC determination of GSH utilize pre-column derivatization of the molecule to produce a chromophore of fluorophore moieties.…”