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Nutrient Deficiencies (Building Blocks Missing) Glutathione synthesis requires specific nutrients: Cysteine : From dietary protein (whey, eggs, poultry)-rate-limiting Glycine : Often deficient in modern diets (need 10-15g/day, most get 3-5g) Selenium : Cofactor for glutathione peroxidase (GPx) , the enzyme that recycles GSSG back to GSH B vitamins (B6, B9, B12): Required for methylation and sulfur metabolism Vitamin C : Recycles oxidized glutathione Deficiency in any of these impairs glutathione production or recycling [5]

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The detailed conditions for the LC-MS analysis of glucuronide formation, transporter substrates, and MPAG are shown in Supplementary Texts S1S3 and Supplementary Table S1

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Cys 2 ), cystine-free (Cys 2 -free), or cystine-free/GSH-supplemented (750 M) medium for the indicated time points were determined