Alcohol soluble carbohydrates (ASCs) comprise one of the most abundant classes of dietary carbohydrates. While it is generally accepted that carbohydrates form a healthy part of the diet, the types and amounts consumed are a source of disagreement among practitioners. In particular, the role of ASCs, commonly referred to as free sugars, and comprising a range of mono-, di-, and oligosaccharides, is an active area of debate. These arguments likely arise from a dearth of chemical structures. Studies have implicated diets high in free glucose, fructose, and other rapidly digested ASCs as being associated with increases in cardiovascular and metabolic diseases, but this belies the full scope of monosaccharides, disaccharides, and oligosaccharides found in food. A growing body of evidence has highlighted the role of indigestible oligosaccharides that are part of "low molecular weight soluble fiber" in promoting human health through gut microbiome-mediated mechanisms. Despite their ubiquitous presence in food and relevance to health, there is a lack of comprehensive, rapid-throughput analytical tools for the absolute quantitation of ASCs. To address this critical gap, we propose a rapid-throughput, highly expandable platform for native Combined Alcohol Soluble CArbohydrate DEtermination (CASCADE) in food. To highlight the robustness of the method across complex matrices, CASCADE analysis was applied to commonly consumed foods including fermented and functionalized foods. The expandability of the platform was demonstrated by the inclusion of sugar alcohols in the analyte panel which are both naturally occurring and added to foods. Overall, we envision CASCADE as a useful tool for the nascent field of precision nutrition, which will necessitate detailed structures of foods.

Retrogradation is a critical step in the physical production of resistant starch. This study aimed to examine the effects of isothermal and temperature-cycled retrogradation on the structural, physicochemical properties, and digestibility of resistant starch type-III (RS3) under various thermal conditions. To create RS3, normal maize starch (NM) and Hylon VII (HAM) were treated by autoclave-microwave and then retrograded at isothermal (4 °C) or various temperature conditions (4/10 °C, 4/20 °C, 4/30 °C, 4/40 °C, and 4/50 °C). We found that temperature-cycled retrogradation possessed greater potential than isothermal retrogradation for producing short-range ordering and crystalline structures of RS3. Also, retrograded starch prepared via temperature cycling exhibited higher double helix content, lower amorphous content, reduced swelling power, and less amylose leaching in water. Furthermore, the starch digestibility was affected by structural alterations, which were more significant in HAM-retrograded starch. While, HAM-4-40 (39.27 %) displayed the highest level of resistant starch (RS).

Carbohydrates comprise the largest fraction of most diets and exert a profound impact on health. Components such as simple sugars and starch supply energy, while indigestible components, deemed dietary fiber, reach the colon to provide food for the tens of trillions of microbes that make up the gut microbiota. The interactions between dietary carbohydrates, our gastrointestinal tracts, the gut microbiome and host health are dictated by their structures. However, current methods for analysis of food glycans lack the sensitivity, specificity and throughput needed to quantify and elucidate these myriad structures. This protocol describes a multi-glycomic approach to food carbohydrate analysis in which the analyte might be any food item or biological material such as fecal and cecal samples. The carbohydrates are extracted by ethanol precipitation, and the resulting samples are subjected to rapid-throughput liquid chromatography (LC)-tandem mass spectrometry (LC-MS/MS) methods. Quantitative analyses of monosaccharides, glycosidic linkages, polysaccharides and alcohol-soluble carbohydrates are performed in 96-well plates at the milligram scale to reduce the biomass of sample required and enhance throughput. Detailed stepwise processes for sample preparation, LC-MS/MS and data analysis are provided. We illustrate the application of the protocol to a diverse set of foods as well as different apple cultivars and various fermented foods. Furthermore, we show the utility of these methods in elucidating glycan-microbe interactions in germ-free and colonized mice. These methods provide a framework for elucidating relationships between dietary fiber, the gut microbiome and human physiology. These structures will further guide nutritional and clinical feeding studies that enhance our understanding of the role of diet in nutrition and health. It is important to understand how carbohydrates are digested-both by human enzymes and by microorganisms present in the gut. This protocol is designed to characterize and quantify food and fecal polysaccharides at the monosaccharide, linkage and polysaccharide level.Analysis is performed by LC-MS/MS. Higher-throughput sample preparation in 96-well plates is possible by using a custom-made clamp to hold the plate lids closed during heating. Complex carbohydrates that are not broken down by human enzymes are food sources for gut microbiota. Toward understanding this process, this protocol describes the quantitative analysis of carbohydrates in food and fecal samples by using LC-MS/MS.

Sweet corn is frequently consumed in the US and contains carbohydrates as major macronutrients. This study examined the effects of blanching, freezing, and canning on carbohydrates in sweet corn. Fresh bi-color sweet corn was picked in the field and processed immediately into frozen and canned samples. Simple sugars, starch, and dietary fiber (DF) (including total DF (TDF), insoluble DF (IDF) and two fractions of soluble DF (SDF)) were measured according to the AOAC methods. Additional glycomic analysis including oligosaccharides, monosaccharide composition of total polysaccharides (MCTP) and glycosidic linkage of total polysaccharides (GLTP) were analyzed using UHPLC-MS. Sucrose is the major simple sugar, and IDF is the main contributor to TDF. Sucrose and total simple sugar concentrations were not altered after blanching or freezing but were significantly reduced in canned samples. Kestose was the only oligosaccharide identified in sweet corn and decreased in all heat-treated or frozen samples. Starch content decreased in frozen samples but increased in canned samples. While two SDF fractions did not differ across all samples, blanching, freezing and canning resulted in increases in TDF and IDF. Six monosaccharides were identified as major building blocks of the total polysaccharides from MCTP analysis. Glucose and total monosaccharide concentrations increased in two canned samples. GLTP was also profoundly altered by different food processing methods. This study provided insights into the changes in the content and quality of carbohydrates in sweet corn after food processing. The data are important for accurate assessment of the carbohydrate intake from different sweet corn products.

目的:探究淀粉—脂质复合物的形成机理.方法:采用12～18个碳链的脂肪酸与脱支/非脱支高直链玉米淀粉复合,利用差示扫描量热仪、X-射线衍射仪、傅里叶红外光谱仪等分析复合物的热特性、结晶结构及有序结构等,探究脂肪酸链长及脱支/非脱支高直链玉米淀粉对淀粉—脂肪酸复合物结构及性质的影响.结果:脱支处理的淀粉与脂肪酸的复合率随碳链的增长而降低,其中月桂酸复合率最高(15.00％);未脱支处理的淀粉与脂肪酸的复合率随碳链的增长先升高后下降,其中软脂酸复合率(13.73％)最高.复合物的热特性、结晶结构、分子有序度等与其复合程度有关,淀粉和脂肪酸复合后,糊化峰值温度升高,热稳定性升高,经复合后,淀粉由B型结晶结构转变为V型.结论:脂肪酸链长及淀粉脱支对淀粉—脂质复合物结构及理化性质具有较大影响.

Pregelatinized starch (PGS) is often used to improve the processing quality of foodstuffs, but little attention has been paid to the effects of different reheating methods and degree of starch gelatinization (DSG) on their rheological and textural properties. In this study, pregelatinized rice starches (RS) with gelatinization degrees ranging from 58% to 100% were prepared via different Rapid Visco Analyser (RVA) heating procedures and reheated in various methods, including high-power microwave (HM), low-power microwave (LM), and water bath. The rheological behavior and textural properties were explored, and the results demonstrated that the consistency, gel strength, hardness, and springiness of PGS in all tested samples decreased significantly after reheating. The storage modulus (G’) of PGS increased dramatically while the thermal stability decreased. Interestingly, the reheating methods possessed various effects on the starch of different DSG.