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As a group of important biopolymers, carbohydrates exist widely in living organisms and play many known and unknown biological roles in life activities via different pathways. Traditionally, carbohydrate polymers, e.g., starch, pectin, polysaccharide, and dietary fiber, are widely applied in industrial applications, e.g., pharmaceuticals, food staff, biofuels, and biomaterials, and a growing understanding and deeper investigation of food carbohydrate are driving the development of natural carbohydrate for novel applications, especially for the treatment of chronic diseases, e.g., hyperlipidemia, obesity, and diabetes. In recent years, the gut microbiota has been considered an important organ and plays a critical role in host health and diseases, and emerging evidence indicates that food carbohydrates are effective for the modulation of gut microbiota. In addition, the modification of carbohydrate polymers could alter or enhance their nutrition properties, which may extend their applications. Notably, the nutrition properties of carbohydrate polymers are dependent on their chemical structures and chain conformations, and thus, the structural identification of carbohydrate polymers and their derivatives is helpful for their development and application in the food and pharmaceutical fields.