Radiometric microbiological assay of vitamin B6: assay simplification and sensitivity study

Modification of a previously developed radiometric microbiological assay for vitamin B6 reduces assay complexity and time. Reduction of enzymatic treatment from 24 to 3 h essentially eliminates one day's time for the analysis of plasma samples. Use of lyophilized Kloeckera brevis cultures eliminates routine subculturing of the test organism, with no significant effect on test results. Modifications in test vial size and total volume in test vials have increased assay sensitivity to a level of 0.25 ng pyridoxine (PN), pyridoxal (PL), or pyridoxamine (PM) per vial level and decreased the amount of medium and labeled substrate (i.e., L-[1-14C]-valine), thus reducing assay cost.