Folate monoglutamate in cereal grains: Evaluation of extraction techniques and determination by LC-MS/MS.

• Folate quantification of cereal grains was optimised and validated. • The effects of mono-/di-/tri-enzyme extraction were attempted. • Di-enzyme treatment with α-amylase and rat conjugase showed better efficiency. • The developed LC–MS/MS method had great reproducibility and sensitivity. Folates are essential micronutrients for human health. To determine the total folate content, the extraction and quantification of seven monoglutamate folate derivatives in cereals (maize, rice, and wheat) were optimised and validated in this study. Di-enzyme treatment with α-amylase and rat conjugase was proved ideal for folate extraction from the cereal grains. The quantification method by liquid chromatography-tandem mass spectrometry was validated based on its matrix effect, linearity, sensitivity, recovery, inter-day and intra-day precision. The limits of detection and quantification of folate derivatives ranged from 0.03–0.88 and 0.1–1.0 μg/100 g among the three cereal samples. The absolute recoveries of most folate derivatives were 72–96 % for these cereal samples, with the exception of dihydrofolate, tetrahydrofolate, and folic acid (44–65 %). The inter-day and intra-day precisions were < 12 % for the three cereals. Analysis of folate content and composition in several cereal grains showed that the total folate levels were approximately 26−37 μg/100 g, with 5-methyl-tetrahydrofolate and 5-methyl-tetrahydrofolate as the dominant. MeFox, an oxidation product of 5-methyltetrahydrofolate, was detected at concentrations 20–39-fold higher than those of total folates in rice and wheat grains. This validated method is an efficient approach for folate determination in cereal crops. [ABSTRACT FROM AUTHOR]