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A simultaneous measurement of nicotinic acid and its direct metabolite, nicotinuric acid, by high-performance liquid chromatography (HPLC) is described. The method uses a Chemcosorb 5-ODS-H (150 × 4.6mm i.d., particle size 5μm) column eluted with 10mM KH2PO4-K2HPO4 buffer containing 5mM tetra-n-butylammonium bromide (pH 7.0)-acetonitrile (100:9, v/v) at a flow-rate of l.Oml/min. The UV detector was set at 260nm. The detection limits for nicotinic acid and nicotinuric acid were 10 pmol (1.23ng) and 10 pmol (1.80ng), respectively, at a signal-to-noise ratio of 5:1. Under these conditions, nicotinuric acid and nicotinic acid were eluted at about 6.3 and 6.9 min, respectively. The technique was used to analyze rat blood and urine after nicotinic acid injection. Total analysis time is about 13 min.

agricultural and biological chemistry

Simultaneous measurement of nicotinic acid and its major metabolite, nicotinuric acid in blood and urine by a reversed-phase high-performance liquid chromatography.