Abstract
The measurement of nicotinamide N-oxide by high-performance liquid chromatography is described. The method used a Chemcosorb 5-ODS-H (150 × 4.5 mm i.d., particle size 5 μm) column eluted with 10 mM potassium dihydrogenphosphate (pH was adjusted to 3.0 by adding concentrated phosphoric acid) containing 0.1 g/l of sodium 1-octanesulfonate-methanol (100:4, v/v) at a flow-rate of 1.0ml/min. The UV detector was set at 260nm. The detection limit for nicotinamide N-oxide was 2pmol (276 pg) at a signal-to-noise ratio of 5:1. Under these conditions, nicotinamide N-oxide was eluted at about 3.2 min. This technique was used to analyze rat urine following effective clean-up steps with chloroform, the total analysis time being about 13 min.
