Abstract
A spectrophotometric recycling assay for the quantitation ofL-phenylalanine (and phenylpyruvate) has previously been reported (Cooperet al., Anal. Biochem.183, 210–214, 1989). The procedure involves the coupling of bacterial phenylalanine dehydrogenase with rat kidney cytosolic glutamine transaminase K. The latter enzyme possesses high affinity for phenylpyruvate. Recycling results in a ≥50-fold increase in sensitivity over that of a conventional spectrophotometric “end point” analysis procedure. The spectrophotometric recycling procedure has now been adapted to the measurement ofL-phenylalanine in microliter quantities of human blood. This procedure is 10 times more sensitive than provided by a commercial kit for the spectrophotometric measurement ofL-phenylalanine in human blood. Moreover, the present results suggest that the recycling procedure adapted for fluorometry will be even more sensitive. By use of suitable dehydrogenases and amino acid aminotransferases it should be possible to quantitate amino acids (in addition to phenylalanine) in small quantities of human blood.
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