Determination of thiols in yeast by HPLC coupled with LTQ-orbitrap mass spectrometry after derivatization with p-(Hydroxymercuri)benzoate.
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.
Rao
Yulan
Y
Center for Instrumental Analysis, China Pharmaceutical University, Nanjing 210009, People's Republic of China.
Xiang
Bingren
B
Bramanti
Emilia
E
D'Ulivo
Alessandro
A
Mester
Zoltan
Z
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006902:Hydroxymercuribenzoates / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D013438:Sulfhydryl Compounds / Q000737:chemistry* / Q000378:metabolism; D015003:Yeasts / Q000737:chemistry* / Q000378:metabolism
D006902:Hydroxymercuribenzoates; D013438:Sulfhydryl Compounds