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Determination of dissolved and particulate thiols in Lake Biwa water and extracted fulvic acids by solid phase extraction followed by HPLC with fluorescence detection
- Rasheduzzaman, Md., Kawaguchi, Mai, Obata, Hajime, Maruo, Masahiro
- Limnology 2018 v.19 no.3 pp. 299-309
- acetylcysteine, antioxidants, bioavailability, biogeochemistry, community structure, cysteine, fluorescence, fulvic acids, glutathione, high performance liquid chromatography, lakes, ligands, metals, phosphine, plankton, reducing agents, solid phase extraction, thiols
- Thiols are important antioxidants that can modulate the bioavailability and biogeochemistry of many soft metals, although their detection remains challenging in both their reduced (R–S) and oxidized (R–S–S–R) forms. Here, a modified biochemical method was applied to determine the levels of dissolved and particulate thiols in Lake Biwa water and extracted Lake Biwa fulvic acids obtained at various depths. This method involves the use of the reducing agent tris(2-carboxyethyl)phosphine and the fluorescent label 7-fluorobenzofurazan-4-sulfonic acid ammonium salt (SBD-F), followed by solid-phase extraction and HPLC with fluorescence detection. Dissolved cysteine (Cys) (2.0–6.0 nM), glutathione (GSH) (2.8–5.1 nM), and N-acetyl-L-cysteine (NAC) (1.6–4.2 nM) were detected throughout the water column but were broadly consistent at depths of 5–20 m. In contrast, abundant levels of particulate cysteine (1.3–3.5 × 10² nM) and glutathione (1.6–3.1 × 10² nM) were detected down to depths of 15 m. The particulate cysteine and glutathione were significantly covariant, and the ratios between them reflected the differences in the plankton community composition and availability of these compounds. This work also studied the concentrations of Cys, GSH and NAC in Lake Biwa fulvic acids (LBFAs) for the first time (at 0 m: cysteine, 0.8 nM; glutathione, 1.6 nM; NAC, 2.5 nM; at 10 m: cysteine, 1.4 nM; glutathione, 0.6 nM; NAC, 1.6 nM). The nanomolar to sub-nanomolar concentrations of the particulate and dissolved Cys, GSH and NAC in the lake indicates that these are an important class of ligands for chalcophile metals and may influence the distribution of plankton communities from the epilimnion to the hypolimnion of the lake.