Development Of New Capillary Zone Electrophoresis Techniques For Analyzing Nature Carbohydrates Composition And Its Application

OBJECT: High performance capillary electrophoresis(HPCE), founded in the1980’s，is a kind of high performance separation technique, which is consideredto be one of the most influencing branch in the fields of analysis and with manyadvantages, such as simplification, high efficiency, reduced analysis time, andsmaller sample volumes. Polysaccharides are concerned with specialpharmacological activity, the monosaccharide composition of polysaccharidesand its activity has a very close relationship, so the most important part is todetermine the monosaccharide composition, in order to provide the basicinformation of polysaccharide. Clearly, a greater understanding of thephysiological effects of carbohydrates will only be possible with the separation,identifcation, and quantifcation of the different classes of carbohydratespresent in foods or drugs. The main purpose of this paper is to establishsuitable food carbohydrate composition of the HPCE analysis methods toprovide an effective method for quality control in food as well aspolysaccharide. METHODS: Crude lycopus lucidus polysaccharides (LPs), jujubepolysaccharides (JPs), honey polysaccharides, Termitomyces albuminosuspolysaccharides (TAs), Panus giganteus polysaccharides(PGs) were extractedand isolated from the palnt materials, by the method of water boiling, ethanolprecipitation, deprotein, lyophilization, ect.. Reducing carbohydrates werederivatized with1-phenyl-3-methyl-5-pyrazolone (PMP) and detected by UV at245nm, the effects of some important factors, such as concentration, the acidityof running buffer, the addition of the organic modifier and the running voltageare explored. Under the optimized conditions, three new HPCE technique for theseparation and analysis of13,11and10carbohydrates achieved. Furthermore,the developed method was applied to determination of the carbohydratescomposition and their molar ratio of LPs, JPs, honey polysaccharides, TAs,PGs, beer and milk.RESULTS AND CONCLUSIONS: A simple, sensitive and specific analyticalmethod has been established for high efficient separation and simultaneouslyhigh sensitive determination of thirteen reducing carbohydrates (includingaldohexose and aldopentoses as well as maltose and lactose). Reducingcarbohydrates were derivatized with1-phenyl-3-methyl-5-pyrazolone (PMP),baseline separated by CE with use of4%methanol modifier in175mM boratebuffer (pH11.0) and15kV running voltage at25℃. The optimized CE methodwas found to be well suited to examine the compositional reducingmonosaccharides of the isolated polysaccharides from L. lucidus and Jujube, andfree mono-and disaccharides in beer and milk. LPs was composed of xylose,arabinose, glucose, ribose, rhamnose, galactose, mannose, glucuronic acid andgalacturonic acid with the molar ratio of1.40：19.64：1.00：0.78：0.27：3.18：0.60：0.24：1.11. JPs was composed of arabinose, glucose, ribose, rhamnose,galactose with the molar ratio of0.08：1.00：0.43：0.06：0.19. It was found thatmaltose and glucose were contained in beer with the molar ratio of2.47：1.00. Milk contained maltose, lactose and glucose with the molar ratio of9.28：3.48：1.00. Quantitative recoveries of the compositional carbohydrates in the sampleswere in the range of93.2-104.0%, and RSD values ranged from2.9%to4.9%.The developed HPCE method proves to be precise and practical for qualitycontrol of reducing carbohydrates, and will provide more highly efficientseparation in food analysis in the future.The results of composition analysis of honey polysaccharides showed that10monosaccharides and maltose as their PMP derivatives were separated withhigh sensitivity, good resolution under the optimized HPCE conditions of175mM of borate buffer at pH11and15kV running voltage at25℃. Thedetermination of10different kinds of honey in a variety of carbohydrate molarratio, there is a different between commodity honey and natural honey in theirreducing carbohydrate. This method is sensitive and cost-effective. It could beuseful for the determination of saccharides in honey.A new HPCE method for the separation and analysis of10monosaccharides was achieved with20min with high sensitivity and baselineresolution under the conditions of175mM of borate buffer at pH11and20kVrunning voltage at25℃. Contrast to the previous experimental research, toimprove the running voltage can greatly improve the separation efficiency. TAswas composed of xylose, arabinose, glucose, galactose, mannose and glucuronicacid with the molar ratio of0.72：1.00：21.69：1.66：26.25：0.72. PGswas composed of xylose, arabinose, glucose, rhamnose, fucose, mannose andglucuronic acid with the molar ratio of0.45：1.00：15.72：0.34：0.24：0.79：0.62. The results demonstrated that the proposed HPCE method was rapid,simple and precise, and it was applicable to the analysis of the compositionalmonosaccharides of fungi polysaccharide.