Open AccessDetermination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate
Author(s) -
Yulan Rao,
Bingren Xiang,
Emilia Bramanti,
Alessandro D’Ulivo,
Zoltán Mester
Publication year - 2010
Publication title -
journal of agricultural and food chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.203
H-Index - 297
eISSN - 1520-5118
pISSN - 0021-8561
DOI - 10.1021/jf903485k
Subject(s) - chemistry , derivatization , tcep , chromatography , glutathione , cysteine , reagent , mass spectrometry , thiol , high performance liquid chromatography , yeast , orbitrap , tris , glycine , biochemistry , organic chemistry , enzyme , phosphine , amino acid , catalysis
A liquid chromatography method with mass spectrometric detection has been developed for the simultaneous determination of six thiols in the sulfur metabolic pathway, including cysteine (Cys), homocysteine (HCys), glutathione (GSH), cysteinyl-glycine (Cys-Gly), gamma-glutamyl-cysteine (Glu-Cys), and S-adenosyl-homocysteine (AdoHcy). Tris(2-carboxyethyl)phosphine (TCEP) was used as the reducing reagent and p-(hydroxymercuri)benzoate (PHMB) as the derivatization reagent. Thiols were extracted from 3 mg of yeast using water in an ultrasonic bath. The absolute detection limits for the compounds studied were in the subpicomole range. It was found that AdoHcy, Cys, GSH, Cys-Gly, Glu-Cys, and very little HCys were present in the selenium-enriched yeast sample studied, and GSH, Glu-Cys, very little AdoHcy, Cys, and Cys-Gly were present in three bakery yeasts.
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