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For the quantification of these metabolites in TIF samples, 5 l samples were extracted in 45 l ice-cold extraction solvent (75% acetonitrile, 25% methanol and 0.1% formic acid) containing isotope-labelled internal standards (36 M 13 C 2 , 15 N-GSH, 0.92 M 13 C 4 , 15 N 2 -GSSG and 10 M D 4 -cystine)

Then Na 2 S (20 M) was injected and the reaction was monitored until reached the maximal absorbance

However, a RCT from this centre, the Folic Acid Supplementation in the Second and Third Trimesters (FASSTT) trial, investigated the effects of extended maternal folic acid supplementation (at the recommended dose of 400 g/d) in trimesters 2 and 3 of pregnancy, and thus beyond the periconceptional period recommended for preventing NTD (Reference McNulty, McNulty and Marshall124)