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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 μL 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 μM [ 13 C 3 , 15 N]-cysteine-NEM, 0.1 μM [ 13 C 2 , 15 N]-GSH-NEM, 2 μM [D 4 ]-Cystine, 4.6 μM [ 13 C 5 , 15 N 2 ]-GSSG, 5 μM [D 4 ]-Hypotaurine, and 40 μM [ 13 C 2 ]-Taurine followed by incubation at 4 °C for 30 min

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This is especially useful in minimising some of the possible side effects of other active ingredients