Detection of carbohydrates using a pre-column derivatization reagent 1-(4-isopropyl) phenyl-3-methyl-5-pyrazolone by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 878; no. 15; pp. 1135 - 1144
• Main Authors: Zhang, Ping, Wang, Zhongfu, Xie, Mingming, Nie, Wanli, Huang, Linjuan
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01-05-2010; Elsevier
• Subjects: 1-(4-Isopropyl) phenyl-3-methyl-5-pyrazolone (PPMP); Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Carbohydrate; Carbohydrates; Carbohydrates - analysis; Chromatography, High Pressure Liquid - methods; Concentration (composition); Derivatives; Derivatization; Electrospray ionization mass spectrometry (ESI-MS); Fundamental and applied biological sciences. Psychology; General pharmacology; High-performance liquid chromatography (HPLC); Ionization; Linear Models; Linearity; Liquid chromatography; Mass spectrometry; Medical sciences; Monosaccharides; Pharmacology. Drug treatments; Pyrazolones - chemistry; Reproducibility of Results; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization - methods; Spirulina - chemistry; Temperature; PPMP; Rha; High-performance liquid chromatography (HPLC); Derivatization; Fuc; GlcNAc; Carbohydrate; HPLC; CE; GlcA; Electrospray ionization mass spectrometry (ESI-MS); ESI-MS; Xyl; HPAEC-PAD; GalA; GalNAc; GlcN; Ara; NMR; PMP; Gal; Man; FACE; GalN; 1-(4-Isopropyl) phenyl-3-methyl-5-pyrazolone (PPMP); Glc; Precolumn; HPLC chromatography; Pyrazole derivatives; Detection; Mass spectrometry; Electrospray
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.03.027

A reagent, 1-(4-isopropyl) phenyl-3-methyl-5-pyrazolone (PPMP) has been synthesized and used for high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) determination of pre-column labeled carbohydrates. Monosaccharides have been quantitatively converted into mono-PPMP-labeled derivatives with 28% aqueous ammonia as a catalyst at 80
°C during 70
min. Mono-PPMP derivatives have been demonstrated to exhibit better chemical stability than bis-PMP ones. PPMP-labeled mixture of twelve monosaccharides (galactosamine, glucosamine, galacturonic acid, glucuronic acid, galactose, glucose/
N-acetylgalactosamine,
N-acetylglucosamine, xylose, arabinose, mannose, fucose, and rhamnose) has been well separated by a reverse-phase HPLC and detected by on-line ESI-MS method under optimized conditions. The data on characteristic fragment ions of the 13 PPMP-labeled monosaccharides with MS
2 data have been collected. The suggested method exhibits good linearity (correlation coefficients
>
0.9975) between the peak areas and the concentration of monosaccharides in a broad concentration range and good reproducibility (RSD
<
3.19%). The developed method has been successfully applied to analyze the monosaccharide composition of natural
Spirulina polysaccharide SPPB-1.