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A simplified HPLC method for rapid determination of folates in yeast with ultraviolet and fluorescence detection without sample purification has been developed. By use of the column Aquasil C(18), specially designed for polar analytes, and gradient elution, it was possible to separate and determine five folate derivatives: tetrahydrofolate, 5-methyltetrahydrofolate, and 5-formyltetrahydrofolate with fluorescence detection, and 10-formylfolic acid and folic acid with ultraviolet detection. The sample preparation required only a small amount of dry yeast (25-50 mg) and included an extraction of folates by heat treatment and deconjugation of folate polyglutamates to monoglutamates with the use of rat serum conjugase. Validation involved investigation of matrix effects, determination of recovery by standard addition method, repeatability, and stability tests. The dominating folate forms in commercial dry baker's yeast were found to be tetrahydrafolate and 5-methyltetrahydrofolate with a total folate content of 2890 microg/100 g (63.4 nmol/g). The simplicity of the method makes it suitable for folate screening studies of different yeast strains.

The B-vitamin folate has specific tasks as a one-carbon (C1) group supplier in the building and repair of DNA and RNA as well as in the methylation of homocysteine to methionine. Folate occurs in all living cells as a dynamic pool of several interconvertible forms carrying different C1 groups. Along the food chain, this dynamic pool of folates constantly changes due to either enzymatic or chemical interconversions during food processing and storage. These interconversions make it difficult to determine individual folate forms in foods. The formyl folates, the second most predominant forms of food folates, after 5-methyltetrahydrofolate, are particularly prone to interconvert at low pH. Today, this knowledge is often neglected, leading to risks for analytical underestimation of formyl folates. The purpose of the review is to explore the stability and interconversions of formyl folates in foods as well as to analyze the pitfalls in the determination of formyl folates.