Determination of monosaccharide composition of Streptococcus thermophilus exopolysaccharides using high performance anion exchange chromatography with pulsed amperometric detector
NIE Caiqing, AI Lianzhong, XIONG Zhiqiang, ZHANG Hui*
(Shanghai Engineering Research Center of Food Microbiology,School of Medical Instrument and Food Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China)
Abstract: This work aimed to establish a stable method to determine monosaccharide composition of Streptococcus thermophilus exopolysaccharides by anion exchange chromatography. The gradient elution procedure was optimized with different concentration and the different deproteinization method was analyzed. The optimal chromatographic conditions were as follows: CarboPac PA20 column (3 mm × 150 mm) was used with elute rate at 0.5 mL/min, the eluent was 1.75 mmol/L NaOH for the first 20 min, followed by a linear NaAc gradient (50-200 mmol/L) with fixed 1.75 mmol/L NaOH for the next 10 min to analyze the neutral with 1.75 mmol/L NaOH. Based on the optimal method, twelve mixed monosaccharide standards were separated. The correlation coefficients of each standard were greater than 0.99, the limits of detection were 2.5-21.1 μg/L, and the recoveries ranged were 77.3%-118.5%. In addition, the optimal deproteinization method was trichloroacetic acid. The monosaccharide composition of exopolysaccharides from four S. thermophilus strains were analyzed with optimized method. The results revealed that the exopolysaccharides were mainly composed of galactosamine, rhamnose, glucosamine, galactose, glucose, mannose, galacturonic acid, and glucuronic acid. This study provides a reliable method for the monosaccharide composition analysis of exopolysaccharides from S. thermophilus.
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2021, Vol. 47 
