Preparation of 1-(2-naphthyl)-3-methyl-5-pyrazolone as pre-column derivatization reagent for separation and determination of saccharides using high performance liquid chromatography-mass spectrometry

1-(2-Naphthyl)-3-methyl-5-pyrazolone (NMP) was prepared and used as pre-column derivatization reagent for the separation of 8 saccharides on a Hypersil ODS 2 column (4.6 mm × 200 mm, 5 μm) using a high performance liquid chromatograph (HPLC) equipped with a diode array detector (DAD). Saccharide derivatives were identified using electrospray ionization-mass spectrometry (ESI-MS) in positive ion mode. It was found that NMP reacted easily with reductive saccharides in the presence of 17% ammonia at 70°C. All linear correlation coefficients for saccharide derivatives were higher than 0.9985. The detection limits (at signal-to-noise of 3:1) for saccharide derivatives ranged from 0.58 to 1.1 pmol. The characteristic fragment ions of the NMP derivatized saccharides, especially the ion at m/z 473, exhibited high regularity for the identification of saccharides mixture. The established method was sensitive and reproducible for the determination of saccharides and was successfully used in the determination of the monosaccharide composition in the rape pollen polysaccharide.