Determination of folacin in foods and other biological materials

The objective of most methods for determination of folates in foods and other biological materials is to estimate the total folacin content of the sample. Because folacin comprises a diverse group of related compounds exhibiting similar biological activity, the analytical method must be capable of measuring all of the folates. Methods have been developed for separation of folates in their monoglutamyl form by using anion-exchange, paired-ion reverse phase, or conventional reverse phase liquid chromatography (LC). The application of these separations to determination of folates in foods and other biological materials has been limited largely by the need for development of adequate preparative methods and sufficiently sensitive and specific detection procedures. Although LC with ultraviolet absorption detection has been successful in certain limited applications, the development of fluorometric detection methods has permitted LC determination of folates in a wide range of materials. Tetrahydrofolic acid and its substituted derivatives are detected by monitoring their native fluorescence in an acid mobile phase, while folic acid and certain other folates are measured by using an oxidative post-column fluorogenic derivatization system. Methods also have been developed for determination of the polyglutamyl chain length distribution of folates in biological materials. In total, these procedures permit a direct determination and characterization of folacin compounds.