2.4.1. Folate measurement
2.4.1.1. Total folates content
Total folate content was determined by HPLC with fluorimetric detection. After extraction all folate vitamers were deconjugated into mono and diglutamate, reduced and methylated into 5-CH3-H4folates. The latter were purified from the extract by affinity chromatography using Folate Binding Protein, and quantified by RP-HPLC with fluorimetric detection (RF-1AXL, Shimadzu Inc., Kyoto, Japan). For experimental details, see Delchier et al. (2013).