Microbiological analysis of 5-formyltetrahydrofolic acid and other folates using an automatic 96-well plate reader.
The growth of auxotrophic bacteria remains the method of choice for the determination of biologically active folate metabolites in plasma. This report describes a microbiological assay for folates adapted to use disposable 96-well plates and an automatic plate reader. The modifications in the assay decreased reagent costs and made the analysis of hundreds of samples per day possible with a sensitivity limit of 10 fmol of (6S)-5-formyltetrahydrofolic acid. This limit compares favorably with that of previously reported, more laborious methods. The unnatural 6R diastereomer of 5-formyltetrahydrofolic acid did not interfere with the microbiological assay of the natural 6S diastereomer.
D001322:Autoanalysis; D001681:Biological Assay; D005492:Folic Acid / Q000097:blood*; D006801:Humans; D007780:Lacticaseibacillus casei / Q000187:drug effects* / Q000254:growth & development; D002955:Leucovorin / Q000097:blood*; D010376:Pediococcus / Q000187:drug effects* / Q000254:growth & development; D013237:Stereoisomerism
D005492:Folic Acid; D002955:Leucovorin