Abstract
An ion-exelusion column chromatographic procedure has been described that accomplishes the separation of isoniazid and its metabolites into completely ionized, slightly ionized and neutral derivatives. By applying specific colorimetric and enzymatic techniques to the column fractions, it was possible to measure isoniazid, isoniazid hydrazones of pyruvic acid and α-ketoglutaric acid, acetyl isoniazid and isonicotinic acid derivatives in urine of human subjects ingesting isoniazid. Isonicotinic acid derivatives were separated via an ion-exchange column chromatographic system. This technique combined with colorimetric procedures demonstrated that isonicotinic acid and isonicotinyl glycine were the only derivatives contributing to the isonicotinic acid fraction of isoniazid metabolites. The glucose hydrazone of isoniazid, diisonicotinyl hydrazine, isonicotinic acid amide, N-methyl isonicotinic acid and isonicotinic acid N-oxide were not detected as urinary metabolites of isoniazid in man.
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