Rapid Quantification of Functional Carbohydrates in Food Products


Current research on milk bioactive components, including complex oligosaccharides, stimulated development of novel milk-based ingredients; this in turn sparked the development of methods that are simultaneously simple and sensitive. Oligosaccharides and glycoproteins present interesting health benefits, including antibacterial and antiviral effects, stimulation of the immune system, and participation in the establishment of a balanced gut microbiome in infants. This work describes the application of a simple and rapid method—Total Carbohydrate Assay—to the determination of functional carbohydrate content in various dairy-based functional products. The miniaturization and optimization of the carbohydrate quantification on microplates afforded good repeatability and sensitivity. The optimized method consumed only minimal amounts of reagents and samples, and carbohydrates were detected in the range from 0 - 20 μg. This assay was successfully applied to determine the content of complicated oligosaccharide mixtures and N-glycans in dairy-derived products. Several complementary analytical techniques were applied to confirm the results. This method is faster and far less expensive than mass spectrometry and it gives a general picture of complex carbohydrate concentrations for instances in which detailed data are not required as needed for research in discrete differences among various biological samples. The ability to quantify glycans in novel food products will provide a unique understanding of the potential of these novel ingredients for use by the food industry.

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Parc, A. , Lee, H. , Chen, K. and Barile, D. (2014) Rapid Quantification of Functional Carbohydrates in Food Products. Food and Nutrition Sciences, 5, 71-78. doi: 10.4236/fns.2014.51010.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] G. V. Coppa, O. Gabrielli, P. Pierani, C. Catassi, A. Carlucci and P. L Giorgi, “Changes in Carbohydrate Composition in Human Milk over 4 Months of Lactation,” Pediatrics, Vol. 91, No. 3, 1993, pp. 637-641.
[2] C. Kunz, S. Rudloff, W. Baier, N. Klein and S. Strobel, “Oligosaccharides in Human Milk: Structural, Functional, and Metabolic Aspects,” Annual Review of Nutrition, Vol. 20, No. 1, 2000, pp. 699-722.
[3] B. Liu and D. S. Newburg, “Human Milk Glycoproteins Protect Infants Against Human Pathogens,” Breastfeeding Medicine, Vol. 8, No. 4, 2013, pp. 354-362.
[4] R. G. LoCascio, M. R. Ninonuevo, S. L. Freeman, D. A. Sela, R. Grimm, C. B. Lebrilla, D. A. Mills and J. B. German, “Glycoprofiling of Bifidobacterial Consumption of Human Milk Oligosaccharides Demonstrates Strain Specific, Preferential Consumption of Small Chain Glycans Secreted in Early Human Lactation,” Journal of Agricultural Food Chemistry, Vol. 55, No. 22, 2007, pp. 89148919.
[5] S. Thurl, M. Munzert, J. Henker, G. Boehm, B. MullerWerner, J. Jelinek and B. Stahl, “Variation of Human Milk Oligosaccharides in Relation to Milk Groups and Lactational Periods,” The British Journal of Nutrition, Vol. 104, No. 9, 2010, pp. 1261-1271.
[6] S. Asakuma, T. Urashima, M. Akahori, H. Obayashi, T. Nakamura, K. Kimura, Y. Watanabe, I. Arai and Y. Sanai, “Variation of Major Neutral Oligosaccharides Levels in Human Colostrums,” European Journal of Clinical Nutrition, Vol. 62, No. 4, 2007, pp. 488-494.
[7] C. Kunz, S. Rudloff, A. Hintelmann, G. Pohlentz and H. Egge, “High-pH Anion-Exchange Chromatography with Pulsed Amperometric Detection and Molar Response Factors of Human Milk Oligosaccharides,” Journal of Chromatography B, Biomedical Sciences and Applications, Vol. 685, No. 2, 1996, pp. 211-221.
[8] S. Thurl, B. Müller-Werner and G. Sawatzki, “Quantification of Individual Oligosaccharide Compounds from Human Milk Using High-pH Anion-Exchange Chromatography,” Analytical Biochemistry, Vol. 235, No. 2, 1996, pp. 202-206.
[9] M. Dubois, K. Gilles, J. K. Hamilton, P. A. Rebers and F. Smith, “A Colorimetric Method for the Determination of Sugars,” Nature, Vol. 168, No. 4265, 1951, pp. 167-167.
[10] A. Laurentin and C. A. Edwards, “A Microtiter Modification of the Anthrone-Sulfuric Acid Colorimetric Assay for Glucose-Based Carbohydrates,” Analytical Biochemistry, Vol. 315, No. 1, 2003, pp. 143-145.
[11] M. Monsigny, C. Petit and A.-C. Roche, “Colorimetric Determination of Neutral Sugars by a Resorcinol Sulfuric Acid Micromethod,” Analytical Biochemistry, Vol. 175, No. 2, 1988, pp. 525-530.
[12] M. Barboza, J. Pinzon, S. Wickramasinghe, J. W. Froehlich, I. Moeller, J. T. Smilowitz, L. R. Ruhaak, J. Huang, B. Lonnerdal and J. B. German, “Glycosylation of Human Milk Lactoferrin Exhibits Dynamic Changes during Early Lactation Enhancing Its Role in Pathogenic Bacteria-Host Interactions,” Molecular and Cellular Proteomics, Vol. 11, No. 6, 2012, Article ID: 015248.
[13] D. L. Aldredge, M. R. Geronimo, S. Hua, C. C. Nwosu, C. B. Lebrilla and D. Barile, “Annotation and Structural Elucidation of Bovine Milk Oligosaccharides and Determination of Novel Fucosylated Structures,” Glycobiology, Vol. 23, No. 6, 2013, pp. 664-676.
[14] T. Masuko, A. Minami, N. Iwasaki, T. Majima, S.-I. Nishimura and Y. C. Lee, “Carbohydrate Analysis by a Phenol-Sulfuric Acid Method in Microplate Format,” Analytical Biochemistry, Vol. 339, No. 1, 2005, pp. 69-72.
[15] C. C. Nwosu, D. L. Aldredge, H. Lee, L. A. Lerno, A. M. Zivkovic, J. B. German and C. B Lebrilla, “Comparison of the Human and Bovine Milk N-Glycome via HighPerformance Microfluidic Chip Liquid Chromatography and Tandem Mass Spectrometry,” Journal of Proteome Research, Vol. 11, No. 5, 2012, pp. 2912-2924.
[16] N. Orsi, “The Antimicrobial Activity of Lactoferrin: Current Status and Perspectives,” Biometals, Vol. 17, No. 3, 2004, pp. 189-196.
[17] L. Seganti, A. M. Di Biase, M. Marchetti, A. Pietrantoni, A. Tinari and F. Superti, “Antiviral Activity of Lactoferrin towards Naked Viruses,” Biometals, Vol. 17, No. 3, 2004, pp. 295-299.
[18] D. Legrand, E. Elass, M. Carpentier and J. Mazurier, “Lactoferrin: Lactoferrin: A Modulator of Immune and Inflammatory Responses,” Cellular and Molecular Life Sciences, Vol. 62, No. 22, 2005, pp. 2549-2559.
[19] B. Lonnerdal, “Nutritional and Physiologic Significance of Human Milk Proteins,” The American Journal of Clinical Nutrition, Vol. 77, No. 6, 2003, pp. 1537S1543S.

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