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Tousoulis, Dimitris, et al

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For the quantification of sulfur metabolites in serum, the metabolites in 20 uL of serum were extracted and derivatized by NEM in 80 L of ice-cold extraction solvent (80% MeOH: 20% H2O containing 25 mM NEM and 10 mM ammonium formate, pH = 7.0) which includes stable isotope labeled internal standards (1 M [ 13 C 3 , 15 N]-cysteine-NEM, 0.1 M [ 13 C 2 , 15 N]-GSH-NEM, 2 M [D 4 ]-Cystine, 4.6 M [ 13 C 5 , 15 N 2 ]-GSSG, 5 M [D 4 ]-Hypotaurine, and 40 M [ 13 C 2 ]-Taurine followed by incubation at 4 C for 30 min