The determination of the optical configuration of naturally occurring amino acids using specific enzymes and paper chromatography

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Abstract
A micromethod for detn. of the optical configuration of amino acids in biological fluids is descr. In its present form the only common amino acids to which it has been shown to be applicable are alanine, histidine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophane, tyrosine and valine. The method involves incubation of the fluid with D-amino acid oxidase of sheep''s kidney and with L-amino acid oxidase of snake venom. The extent of the reactions is followed by paper-chromatographic analysis. D-amino acid oxidase, free from amino acids and active enough for the present purposes, was prepared without the need to split off and subsequently to add the coenzyme. The method was applied to acid hydrolysate of casein, to a human plasma ultrafiltrate, to the cerebrospinal fluid from 2 cases of phenyl-ketonuria (phenylpyruvic oligophrenia) and to urine from 2 cases of Fanconi syndrome (adult type), 2 cases in children of Fanconi syndrome with cystinosin, and one case of a syndrome involving renal amino aciduria, not yet fully descr. clinically. No evidence was obtained of the presence in these fluids of D forms of the above 10 amino acids.