QUANTITATIVE-DETERMINATION OF N-ARYLACETOLHYDROXAMIC AND N-ARYLGLYCOLHYDROXAMIC ACIDS IN BIOCHEMICAL REACTION MIXTURES

A high-performance liquid chromatography (hplc) solvent system was developed for use with C18 bonded packings that effected the chromatographic resolution of monoaromatic hydroxamic acids in complex mixtures. The success of this hplc system resulted from the incorporation of an aliphatic hydroxamic acid additive into the elution solvent. This additive, desferal mesylate, suppressed chemisorption effects between the stationary phase of the chromatographic packing and the aromatic hydroxamic acids being analyzed. The detectability limit for acetyl-derived aromatic hydroxamic acids was about 10 ng, while that for glycolyl-derived aromatic hydroxamic acids was about 5 ng. This level of sensitivity was sufficiently low to allow for the direct detection of these components in enzymatic reaction mixutres. By manipulation of the percentage methanol in the aqueous solvent the resolution of either type of hydroxamic acid from other related metabolites was readily accomplished. © 1979.