Abstract
An enzymatic-HPLC procedure for the determination of plasma pyridoxal 5′-phosphate (PLP) has been established. The assay is based on the decarboxylation of l-3,4-dihydroxyphenylalanine using Streptococcus tyrosine decarboxylase apoenzyme, which requires PLP as cofactor. The product of the enzyme reaction, dopamine, is measured by Coulochem electrochemical detection with a series of oxidizing and then reducing electrodes. Trace amounts of PLP in the apoenzyme preparation were removed with the aid of cysteine-sulfinic acid and gel filtration. The detection limit for PLP by this method is 50 p m in plasma.
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