Challenges in the determination of unsubstituted food folates: impact of stabilities and conversions on analytical results.

Tetrahydrofolate is the parent molecule of the folate coenzymes required for one carbon metabolism. Together with other unsubstituted folates such as dihydrofolate and folic acid, tetrahydrofolate represents the third pool of dietary folates following 5-methyltetrahydrofolate and formyl folates. Low intake of dietary folates and poor folate status are common problems in many countries. There is a critical need for reliable methods to determine folate in foods to accurately estimate folate intakes in populations. However, current values for folates in foods in databanks are often underestimated due to the high instability of several folate forms, especially tetrahydrofolate. The present review highlights the occurrence of unsubstituted folates in foods and their oxidation mechanisms and chemical behavior as well as interconversion reaction between tetrahydrofolate and 5,10-methylenetetrahydrofolate. The review shows also the important role of antioxidants in protecting folates during analysis and describes strategies to stabilize unsubstituted folates throughout all steps of the analytical procedure.