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A novel analytical method using liquid chromatography coupled to isotope ratio mass spectrometry (LC/IRMS) was developed for measuring the fractional synthesis rate (FSR) of glutathione (GSH) in neonates after infusion of [1-13C]-glycine as a tracer. After transformation of GSH into GSSG, its dimeric form, the intra-erythrocytic concentration and 13C-isotopic enrichment of GSH were determined using 200 µL of blood. The results showed that, using LC/IRMS, the concentration (range of µmol/mL) was reliably measured using norvaline as internal standard with precision better than 0.1 µmol/mL.