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.
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 degrees 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.
Zhang
Ping
P
Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, Life Science College, Northwest University, No. 229, Tibai North Road, Xi'an 710069, China.
Wang
Zhongfu
Z
Xie
Mingming
M
Nie
Wanli
W
Huang
Linjuan
L
D002241:Carbohydrates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016014:Linear Models; D047069:Pyrazolones / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053545:Spirulina / Q000737:chemistry; D013696:Temperature
C551032:1-(4-isopropyl)phenyl-3-methyl-5-pyrazolone; D002241:Carbohydrates; D047069:Pyrazolones