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Simple and sensitive quantification of glutathione hydropersulfide alkylated using iodoacetamide by high-performance liquid chromatography with post-column derivatization.
- Source :
-
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2021 Jan 15; Vol. 1163, pp. 122516. Date of Electronic Publication: 2020 Dec 25. - Publication Year :
- 2021
-
Abstract
- A novel analytical method was developed for the quantification of glutathione hydropersulfide (G-SSH) in biological samples by high-performance liquid chromatography (HPLC) with post-column derivatization. G-SSH was treated with iodoacetamide as an alkylating agent for 5 min at 37 °C, and the resultant acetamide-labeled G-SSH (G-SS-acetamide) was subjected to HPLC. After separation on a reversed-phase column, G-SS-acetamide was quantified by detection using a post-column reaction with orthophthalaldehyde under alkaline conditions. The standard G-SS-acetamide was synthesized through the S-S exchange reaction between oxidized glutathione and 2-mercaptoacetamide. It should be noted that some types of alkylating agents, including N-ethylmaleimide and monobromobimane, cleave the polysulfide chains of polysulfides that consist of glutathione, resulting in the production of alkylated G-SSHs. We confirmed that iodoacetamide did not enhance the cleavage of acetamide-labeled glutathione trihydropersulfide (G-SSS-acetamide). The lowest quantification limit was estimated to be 25 nM for G-SS-acetamide. This method can be useful for studying the dynamics of sulfane sulfur in glutathione-containing matrices.<br /> (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Subjects :
- Cell Line, Tumor
Glutathione analysis
Glutathione chemistry
Glutathione metabolism
Humans
o-Phthalaldehyde chemistry
Alkylating Agents chemistry
Chromatography, High Pressure Liquid methods
Disulfides analysis
Disulfides chemistry
Disulfides metabolism
Glutathione analogs & derivatives
Iodoacetamide chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1873-376X
- Volume :
- 1163
- Database :
- MEDLINE
- Journal :
- Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
- Publication Type :
- Academic Journal
- Accession number :
- 33429128
- Full Text :
- https://doi.org/10.1016/j.jchromb.2020.122516