Separation and quantitation of the seven forms of vitamin B-6 in plasma and 4-pyridoxic acid in urine of adolescent girls by reverse phase high performance liquid chromatography

The vitamin B-6 status of seemingly healthy adolescent girls was determined using several accepted and proposed parameters in an effort to establish guidelines for status evaluation. HPLC-derived plasma B-6 vitamer [pyridoxal phosphate (PLP), pyridoxine phosphate (PNP), pyridoxamine phosphate (PMP), pyridoxal (PL), pyridoxine (PN), and pyridoxamine (PM)] and 4-pyridoxic acid (4-PA) concentrations and urinary 4-PA levels of 28 white adolescent females, 12-15 years, having radiomonitored plasma PLP concentrations and coenzyme stimulation of erythrocyte alanine aminotransferase activities indicative of adequate status were determined. Mean daily vitamin B-6 and protein intakes of the subjects were 1.48 mg and 78.3 g, respectively. The ranges for plasma B-6 vitamer and 4-PA concentrations for these subjects which had seemingly adequate vitamin B-6 status were as follows: (nmol/L) PLP, 40.9-122.2; PNP, 0-16.1; PMP, 0-8.1; PL, 0- 15.0; PN, 0-21.9; PM, 0-17.8; and 4-PA, 0-55.7. PLP was the predominant plasma B-6 vitamer as well as being the only vitamer found in plasma of all subjects. Urinary 4-PA concentrations of the girls ranged from 0.11-2.50 pmol/mmol creatinine. The B-6 vitamer values of these white adolescent girls should be of use in the establishment of normal ranges for vitamin B-6 status parameters. HPLC methodologies seem to be advantageous for the rapid and accurate assessment of vitamin B-6 status.