Development of a derivatization method for the quantification of hydrogen sulfide and its application in vascular calcification rats.
J Chromatogr B Analyt Technol Biomed Life Sci
; 1055-1056: 8-14, 2017 Jun 15.
Article
en En
| MEDLINE
| ID: mdl-28437716
ABSTRACT
Hydrogen sulfide (H2S) plays major functional and structural roles in diverse physiological functions and the pathogenesis of a variety of disorders in biological matrices. The significance of H2S has prompted the development of sensitive and selective methods to determine its concentration in biological samples. The fluorescent reagent monobromobimane (MBB) has been widely used to measure various thiol-containing species through alkylation. MBB may prevent the oxidation of sulfide and the reaction of sulfide with several different species (such as superoxide radicals, hydrogen peroxide and peroxynitrite). An isomers of MBB, 3-(bromomethyl)-2, 6, 7-trimethyl-1H, 5H-pyrazolo [1,2-a] pyrazole-1, 5-dione (MMB), is cheaper than MBB and its use in the analysis of H2S has not previously been reported. In the present study, we compared the derivatization reactions of hydrogen sulfide with MMB and MBB and developed a sensitive method to quantify H2S in blood. In our method, H2S was incubated in the dark with excess MMB in 0.1M Tris-HCl buffer (pH 10.1) at 50°C for 120min. 50µL aliquots of the derivatized product were analyzed using HPLC system with gradient elution of 0.1% (v/v) formic acid-acetonitrile. The limit of detection for the derivatized product was 0.03nmol/mL. The derivatization reaction was suitable for detecting low concentrations of H2S. The derivate product is stable over time, permitting batch storage and analysis.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Cromatografía Líquida de Alta Presión
/
Calcificación Vascular
/
Sulfuro de Hidrógeno
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
J Chromatogr B Analyt Technol Biomed Life Sci
Asunto de la revista:
ENGENHARIA BIOMEDICA
Año:
2017
Tipo del documento:
Article
País de afiliación:
China