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A new colorimetric method for the determination of hydroxamic acid is described. Hydroxamic acid was oxidized quantitatively by iodine to produce nitrous acid, which was thereafter determined according to the diazocoupling reaction. This method is sensitive to as little as 5 nmol of hydroxamic acid, and the calibration curve is linear up to 50 nmol. Using this method, acyl-CoAs were determined after conversion to hydroxamic acid by the addition of hydroxylamine. The present method is applicable to the determination of free fatty acids which are activated by acyl-CoA synthetase.
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