Analysis of B6 vitamers in plasma by reversed-phase column liquid chromatography

The seven vitameric forms of B6 can be measured in biologic fluids by high-performance liquid chromatography. Use of a reversed-phase column, with optimized solvent and buffer conditions, combined with fluorometric detection, allowed linear detection of vitamers from 0.4 to 20 ng. All vitamers from plasma, urine or tissue extracts can be analyzed within 50 min. Reproducibility and recovery studies indicate a selective and sensitive procedure, which greatly enhances studies on vitamin B6 metabolism.