Abstract
A high performance liquid chromatographic method involving postcolumn photochemical reaction and fluorometric detection has been developed for the determination of isonicotinic acid, acetylisoniazid and isoniazid in human urine. These compounds are separated by reversed phase chromatography using 0.07 M phosphate buffer (pH 6.8) containing 100 mM hydrogen peroxide as a mobile phase. The compounds in the column effluent are irradiated with ultraviolet light to give fluorescence. The fluorescence is monitored with excitation at 316nm and emission at 418 nm. The calibration curves for isonicotinic acid, acetylisoniazid and isoniazid are linear over the ranges of 0.1–120, 1.0–180 and 0.5–200 ng, respectively. The mean recoveries of isonicotinic acid, acetylisoniazid and isoniazid from urine are more than 92%. This method can be applied for acetylator phenotyping.
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