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Chinese Journal of Chromatography
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Chinese Journal of Chromatography  2007, Vol. 25 Issue (1): 75-79    DOI:
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Analysis of Monosaccharides and Uronic Acids in Polysaccharides
by Pre-column Derivatization with p-Aminobenzoic Acid
and High Performance Liquid Chromatography

HAO Guitang1, CHEN Shangwei2, ZHU Song2, YIN Hongping3, DAI Jun2, CAO Yuhua1

1.School of Chemistry and Material Engineering of Southern Yangtze University, Wuxi 214036, China;
2.Testing & Analysis Center of Southern Yangtze University, Wuxi 214036, China; 3.School of
Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
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Abstract 

An ion-pair reversed-phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous determination of carbohydrate and uronic acids was developed. p-Aminobenzoic acid (p-AMBA) was used for pre-column derivatization of the analytes, enabling fluorescence (λex=313 nm, λem=358 nm) or ultraviolet (UV at 303 nm) detection. Reaction conditions such as reaction temperature and reaction time were optimized. Atlantis dC18 column with hydrophilic end capping was selected for the separation of derivatives. Effects of mobile phase compositions such as ion pairs and their concentrations and pH on the retention behaviors and separation results of 9 monosaccharides and 2 uronic acids were investigated. Derivatives of fructose, galactose, glucose, mannose, xylose, arabinose, ribose, galacturonic acid, fucose, glucuronic acid and rhamnose were separated within 42 min, applying tetrabutyl ammonium hydrogen bisulfate (TBAHSO4) as the ion pair reagent. The detection limits were between 3.38×108 mol/L and 176×108 mol/L for fluorescence detection and between 2.55×107 mol/L and 13.4×107mol/L for UV detection. Good linearities were obtained with correlation coefficients (r2) above 0.99. The relative standard deviations (RSDs) of the peak area of the derivatives in 1251 h after derivatization were from 2.5% to 3.9%. This method has been applied for the determination of mono-/disaccharides and uronic acids in spirulina polysaccharide after dissolved in trifluoroacetic acid solution (2 mol/L). The results showed this method is suitable for the analysis of monosaccharide compositions in polysaccharides.

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Articles by authors

HAO Guitang

CHEN Shangwei
ZHU Song
YIN Hongping
DAI Jun
CAO Yuhua

Key wordspre-column derivatization   high performance liquid chromatography (HPLC)   p-aminobenzoic acid   monosaccharide   uronic acid   polysaccharides      
Received: 2006-05-16; Published: 2007-01-30
Corresponding Authors: DAI Jun   
Cite this article:   

HAO Guitang, CHEN Shangwei, ZHU Song et al.

Analysis of Monosaccharides and Uronic Acids in Polysaccharides
by Pre-column Derivatization with p-Aminobenzoic Acid
and High Performance Liquid Chromatography

[J]. Chinese Journal of Chromatography, 2007, 25(1): 75-79.
 
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