Determination of plasma pyridoxal phosphate levels using a modified apotryptophanase assay

The development of a simple non-isotopic enzymatic assay for plasma pyridoxal phosphate (PLP) using a modified apotryptophanase procedure is reported. Recovery of added PLP from plasma was 86% and the within-run precision of the assay was 6.5%. Between-run precision was 7.5%. Concentrations of cupric ions as low as 20 mumol/L severely inhibited apotryptophanase activity and this effect of copper was almost completely prevented by the addition of EDTA. The reference plasma PLP range obtained by the apotryptophanase method was found to be between 5 and 50 micrograms/L plasma, with a mean of 17 +/- 10 micrograms/L plasma. The assay requires minimum technical skill and, with the aid of a multisample reaction rate analyser, more than 50 samples could easily be handled within a working day.