Naturally-occurring folates in foods: Method development and analysis using liquid chromatography–tandem mass spectrometry (LC–MS/MS)

An accurate method for detecting and quantifying both synthetic (folic acid) and naturally-occurring folates in foods is described. A system capable of analysing the five most commonly occurring folates (pteroylglutamic acid, 5-methyltetrahydrofolate, tetrahydrofolate, 10-formylfolate and 5-formyltetrahydrofolate) in 20 min using liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed. Quantification of folates was performed using 13C labelled internal standards. This paper outlines the development of a comparatively fast LC–MS/MS method, method validation using commercially available folate standards and establishment of the method’s suitability for quantification using selected reaction monitoring (SRM) mass spectrometry. The application of the system was verified by analysing several certified reference materials and comparing results with certified values as determined by microbiological assay. LC–MS/MS promises to be an ideal tool for the quantitative analysis of folates in food.