Carbohydrates, analysis

Sugar analysis by hplc has advanced greatly as a result of the development of columns specifically designed for carbohydrate separation. These columns fall into several categories. (/) Aminopropyl-bonded siHca used in reverse-phase mode with acetonitrile—water as the eluent. (2) Ion-moderated cation-exchange resins using water as the eluent. Efficiency of these columns is enhanced at elevated temperature, ca 80—90°C. Calcium is the usual counterion for carbohydrate analysis, but lead, silver, hydrogen, sodium, and potassium are used to confer specific selectivities for mono-, di-, and... 

The purpose of the experiment is to illustrate the application of derivatisation in the analysis of sugar and related substances by gas-liquid chromatography. The silylation method described is an almost universal derivatisation procedure for carbohydrate analysis by GC.79... 

A typical application of the RI detector is in carbohydrate analysis. Carbohydrates do not adsorb in the UV, do not ionize and although fluorescent derivatives can be made, the procedure is tedious. Consequently, the RI detector can be ideal for detecting such materials and an example of such an application is shown in figure 18. 

Lee, YC (1990) High-performance anion-exchange chromatography for carbohydrate analysis. AnalBiochem 189 151-162... 

Cotte, J. F., Casabianca, H., Chardon, S., Lheritier, J., and Grenier-Loustalot, M. F. (2003). Application of carbohydrate analysis to verify honey authenticity. /. Chromatogr. A 1021, 145-155. 

Low, N. H., Brisbane, T., Bigam, G., and Spoms, P. (1988). Carbohydrate analysis of western Canadian honeys and their nectar sources to determine the origin of honey oligosaccharides. /. Agric. Food Chem. 36, 953-957. 

White, J. W. (1980). Detection of honey adulteration by carbohydrate analysis. /. Assoc. Ojfic. Anal. Chem. 63,11-18. 

De Ruiter, G. A., Schols, H. A., Voragen, A. G. J., and Rombouts, F. M., Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods, Anal. Biochem., 207, 176, 1992. 

Das, H., Jayaraman, V., and Bhattacharya, J., Carbohydrate analysis of bradyrhizobial (NC 92) lipopolysaccharides by high-performance-anion exchange chromatography with pulsed amperometric detection, Biosci. Rep., 19, 219, 1999. 

Although a number of assays and technologies are available to characterize and test protein molecules, such as peptide mapping, protein sequencing, carbohydrate analysis, electrophoresis, ELISA, and mass spectroscopy, they are not as definitive as the methods used for small molecule drugs. Hence, the test for similarity is not as well defined in the case of proteins. However, as... 

B. Monitoring rMAb Charge Heterogeneity CARBOHYDRATE ANALYSIS BY CE-LIF... 

One of the applications of CE for carbohydrate analysis that has shown increase acceptance in the biotechnology industry is the relative distribution of the N-linked oligossacharides on the conserved asparagines residue on the Fc portion of rMAbs. The advantages of CE over traditional methods include enhanced separation efficiencies, shorter analysis time, and improved robustness. The oligosaccharides present on a monoclonal antibody are asialo-N-linked complex biantennary structures with a core fucose, typical of the structures produced in the Chinese hamster ovary (CHO) cells. The main forms share the fucosylated branched core structure but vary in their terminal galactose occupancy ... 

CE has recently emerged as a powerful tool in carbohydrate analysis with enhanced resolution for isobaric isomers, shorter analysis times, and high sensitivity with EIE detection, as well as better assay reproducibility and robustness over the traditional methods. 

Townsend, R. R. (1995). Carbohydrate Analysis High-Performance Liquid Chromatography and Capillary Electrophoresis, Elsevier, New York. 

Glycoprotein Carbohydrate Analysis. Gels from PAGE were fixed with iso-propanol-acetic acid-water (25 10 65 v v v), stained with 0.2% thymol in the fixing solution, and washed two or three times with the same fixing solution. This was followed by staining with concentrated H2S04-ethanol (4 1 v/v) at 35°C for 2-3 h (72). 

White CA, Kennedy IF (1986) Oligosaccharides. In Chaplin ME, Kennedy IF (eds) Carbohydrate Analysis. A Practical Approach. IRL Press, Oxford, UK p 37 Okabe H, Kawasaki T (1970) Structures of Pharbitic Acids C and D. Tetrahedron Lett 36 3123... 

Historically, R1 detector was widely applied to carbohydrates analysis [52,55,59,66-71], Other approaches involve the use of ED permitting a high sensitivity and selectivity even in the presence of complex matrices further improvement in this method is the use of pulsed amperometric detector [69,70,72-78],... 

Many authors have commented upon the problems associated with the hydrolytic step, which has been considered to be the main source of loss in carbohydrate analysis.19 The fact that, in determining uronic acids, differing results were obtained, depending on whether hydrolysis or methanolysis was employed, led others20 to conclude that... 

Tanowitz, B. D. and Koehler, D. L. (1986). Carbohydrate analysis of floral and extra-floral nectars in selected taxa of Sansevieria (Agavaceae). Annals of Botany 58 541-545. 

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