1991 Volume 7 Issue 5 Pages 733-736
A high performance liquid chromatographic method involving postcolumn ultraviolet (UV) irradiation and fluorometric detection has been developed for the simultaneous determination of nicotinic acid and nicotinamide in human serum. These compounds are separated by reversed-phase chromatography using a 0.07M potassium dihydrogenphosphate solution (pH 4.5) containing 75mM hydrogen peroxide and 5μM copper(II) sulfate as a mobile phase. The compounds in the column effluent are irradiated with ultraviolet light to produce fluorescence. This fluorescence is monitored with excitation at 322nm and emission at 380nm. The calibration curves for nicotinamide, nicotinic acid and N1-methylnicotinamide are linear over the ranges of 0.6-136, 0.7-166 and 5.5-220ng, respectively. The mean recoveries of nicotinamide and nicotinic acid from serum are more than 93%.
