Simultaneous micro-determination of nicotinamide and its major metabolites, N1-methyl-2-pyridone-5-carboxamide and N1-methyl-4-pyridone-3-carboxamide, by high-performance liquid chromatography

J Chromatogr. 1988 Jan 22;424(1):23-8. doi: 10.1016/s0378-4347(00)81072-5.

Abstract

A simultaneous micro-determination of nicotinamide and its major metabolites, N1-methyl-2-pyridone-5-carboxamide (2-py) and N1-methyl-4-pyridone-3-carboxamide (4-py) by high-performance liquid chromatography is described. The method employs a 7-ODS-L (250 mm X 4.6 mm I.D., particle size 7 microns) column eluted with 10 mM potassium dihydrogenphosphate-acetonitrile (96:4, v/v; pH adjusted to 3.0 by the addition of concentrated phosphoric acid) at a flow-rate of 1.0 ml/min. The UV detector was set at 260 nm. The detection limits for nicotinamide, 2-py and 4-py were 10 pmol (1.22 ng), 2 pmol (304 pg) and 2 pmol (304 pg), respectively, at a signal-to-noise ratio 5:1. Isonicotinamide was used as an internal standard. The technique was applied to the analysis of rat and human urines. The total analysis time was ca. 15 min.

MeSH terms

  • Animals
  • Chromatography, High Pressure Liquid
  • Female
  • Humans
  • Male
  • Niacinamide / analogs & derivatives*
  • Niacinamide / analysis*
  • Niacinamide / blood
  • Niacinamide / urine
  • Rats
  • Rats, Inbred Strains
  • Spectrophotometry, Ultraviolet

Substances

  • Niacinamide
  • N(1)-methyl-4-pyridone-5-carboxamide