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Removal of ferrous ions by N,N'-ethylenediaminetetraacetic acid in microbiological assay of pyridoxine.

The application of a general microbiological method for the determination of pyridoxine was satisfactory for most preparations containing pyridoxine, but there were discrepancies in some multivitamin products. The chief cause of these discrepancies was shown to be Fe+2. A modification was made to the method by using N,N′-ethylenediaminetetraacetic acid to sequester Fe+2 before acid hydrolysis of the preparations. By using this modification, 100% of the pyridoxine can be detected compared with 75 or 77% for 20:1 or 12:1 ratios of Fe+2–pyridoxine, respectively.