In vitro digestion characteristics of unprocessed and processed whole grains and their components

Chemical composition and in vitro digestion properties of select whole grains, before and after processing, and their components were measured. Substrates included barley, corn, oat, rice, and wheat. In addition to whole grain flours, processed substrates also were tested as were corn bran, oat bran, wheat bran, and wheat germ. Processing of most substrates resulted in higher dry matter and digestible starch and lower resistant starch concentrations. Dietary fiber fractions varied among substrates with processing. Digestion profiles for most substrates correlated well with their chemical composition. Corn bran and rice substrates were the least fermentable. Extrusion rendered barley, corn, and wheat more hydrolytically digestible and barley and oat more fermentatively digestible. Except for corn bran, all components had greater or equal fermentability compared with their native whole grains. Understanding digestion characteristics of whole grains and their components will allow for more accurate utilization of these ingredients in food systems.     

Contents of phenolic acids, alkyl- and alkenylresorcinols, and avenanthramides in commercial grain products

The contents of free and total phenolic acids and alk(en)ylresorcinols were analyzed in commercial products of eight grains: oat (Avena sativa), wheat (Triticum spp.), rye (Secale cerale), barley (Hordeum vulgare), buckwheat (Fagopyrum esculentum), millet (Panicum miliaceum), rice (Oryza sativa), and corn (Zea mays). Avenanthramides were determined in three oat products. Free phenolic acids, alk(en)ylresorcinols, and avenanthramides were extracted with methanolic acetic acid, 100% methanol, and 80% methanol, respectively, and quantified by HPLC. The contents of total phenolic acids were quantified by HPLC analysis after alkaline and acid hydrolyses. The highest contents of total phenolic acids were in brans of wheat (4527 mg/kg) and rye (4190 mg/kg) and in whole-grain flours of these grains (1342 and 1366 mg/kg, respectively). In other products, the contents varied from 111 mg/kg (white wheat bread) to 765 mg/kg (whole-grain rye bread). Common phenolic acids found in the grain products were ferulic acid (most abundant), ferulic acid dehydrodimers, sinapic acid, and p-coumaric acid. The grain products were found to contain either none or only low amounts of free phenolic acids. The content of avenanthramides in oat flakes (26-27 mg/kg) was about double that found in oat bran (13 mg/kg). The highest contents of alk(en)ylresorcinols were observed in brans of rye (4108 mg/kg) and wheat (3225 mg/kg). In addition, whole-grain rye products (rye bread, rye flour, and whole-wheat flour) contained considerable levels of alk(en)ylresorcinols (524, 927, and 759 mg/kg, respectively).     

Extrusion conditions affect chemical composition and in vitro digestion of select food ingredients

An experiment was conducted to determine the effects of extrusion conditions on chemical composition and in vitro hydrolytic and fermentative digestion of barley grits, cornmeal, oat bran, soybean flour, soybean hulls, and wheat bran. Extrusion conditions altered crude protein, fiber, and starch concentrations of ingredients. Organic matter disappearance (OMD) increased for extruded versus unprocessed samples of barley grits, cornmeal, and soybean flour that had been hydrolytically digested. After 8 h of fermentative digestion, OMD decreased as extrusion conditions intensified for barley grits and cornmeal but increased for oat bran, soybean hulls, and wheat bran. Total short-chain fatty acid production decreased as extrusion conditions intensified for barley grits, soybean hulls, and soybean flour. These data suggest that the effects of extrusion conditions on ingredient composition and digestion are influenced by the unique chemical characteristics of individual substrates.     

Fermentability of Oat and Wheat Fractions Enriched in β‐Glucan Using Human Fecal Inoculation

Fermentation by human fecal bacteria of fractions of wheat bran prepared by preprocessing technology were examined and compared with a β‐glucan‐rich oat bran and a purified β‐glucan (OG). The wheat fractions were essentially a beeswing bran (WBA), mainly insoluble dietary fiber, and an aleurone‐rich fraction (WBB) containing more soluble fiber and some β‐glucan (2.7%). The oat bran (OB) had more endosperm and was very rich in β‐glucan (21.8%). Predigestion of WBB and OB to mimic the upper gastrointestinal (GI) tract gave digested wheat bran fraction B (WBBD) and digested oat bran (OBD), respectively. These predigested fractions were fermented in a batch technique using fresh human feces under anaerobic conditions. Changes in pH, total gas and hydrogen production, short chain fatty acids (SCFA), and both soluble and insoluble β‐glucan and other polysaccharide components, as determined from analysis of monosaccharide residues, were monitored. Fractions showed increasing fermentation in the order WBA < WBBD < OBD < OG. Variations in SCFA production indicated that microbial growth and metabolism were different for each substrate. Polysaccharide present in the supernatant of the digests had disappeared after 4 hr of fermentation. Fermentability of oat and wheat β‐glucan reflected solubility differences, and both sources of β‐glucan were completely fermented in 24 hr. Although the overall patterns of fermentation indicated the relative amounts of soluble and insoluble fiber, the anatomical origin of the tissues played a major role, presumably related to the degree of lignification and other association with noncarbohydrate components.     

Whole Grains and Digestive Health

Whole grains contain all parts of the grain: the endosperm, germ, and bran. Whole grains are rich in fermentable carbohydrates that reach the gut: dietary fiber, resistant starch, and oligosaccharides. Most research that supports the importance of grains to gut health was conducted with isolated fiber fractions, rather than whole grains. Whole grains are an important source of dietary fiber and grain fibers such as wheat, oats, barley, and rye increase stool weight, speed intestinal transit, get fermented to short chain fatty acids, and modify the gut microflora. Wheat bran is particularly effective in increasing stool weight; wheat bran increases stool weight by a ratio of 5:1. In contrast, many novel fibers that are easily incorporated into beverages and foods increase stool weight only on a ratio of 1:1. In vitro fermentation studies with whole grains have been published. Carbohydrates of oat bran (rich in β‐glucan) were consumed by bacteria faster than those of rye and wheat brans (rich in arabinoxylan). Grain fibers were fermented more slowly than inulin, causing less gas production. Wheat is particularly high in fructo‐oligosaccharides, while wheat germ is high in raffinose oligosaccharides. Some in vivo studies show the prebiotic potential of whole grains. Whole grain breakfast cereal was more effective than wheat bran breakfast cereal as a prebiotic, increasing fecal bifidobacteria and lactobacilli in human subjects. Wheat bran consumption increased stool frequency. Thus, the gut enhancing effects of cereal fibers are well known. Limited data exist that whole grains alter gut health.     

Metabolite production during in vitro colonic fermentation of dietary fiber: analysis and comparison of two European diets

Metabolite production and antioxidant released during colonic fermentation of naturally occurring dietary fiber (DF) from two European diets (Mediterranean and Scandinavian) were determined. With this aim, DF and associated components were isolated from both whole diets, as well as from cereals and fruits and vegetables comprising the diets. DF was used as substrate for colonic fermentation in a dynamic in vitro model of the colon, samples were collected, and fermentation metabolites were analyzed. Statistical differences between samples were observed in the concentrations of short-chain fatty acids and ammonia and in the ratio acetate/propionate/butyrate. Whole grain cereal DF generated a larger amount of propionate than refined flour cereal DF. Fruit and vegetable DF generated higher amounts of butyrate than cereal DF. Most antioxidant compounds were released from DF during in vitro colonic fermentation. It is concluded that different sources of DF may play a specific role in health maintenance mediated by metabolites produced during colonic fermentation.     

Effects of Different Cereal Fibers on Cholesterol and Bile Acid Metabolism in the Syrian Golden Hamster

This study examined the effects of various cereal fibers and various amounts of β-glucan on cholesterol and bile acid metabolism. Hamsters were fed semisynthetic diets containing 0.12% cholesterol, 20% fat, and either 16% total dietary fiber (TDF) from wheat bran (control) or 10% TDF from oat bran, 13% TDF from oat bran concentrate or barley grains, 16% TDF from oat fiber concentrate, barley flakes, or rye bran. After five weeks, plasma total cholesterol and liver cholesterol concentrations were significantly lower (20 and 50%, respectively) only in hamsters fed rye bran. Diets containing any of the oat ingredients or barley had no effect on total cholesterol. Changes in the pattern of biliary bile acids occurred in hamsters fed 16% TDF from barley flakes or 10% TDF from oat bran. Hamsters fed rye bran had a significantly higher fecal bile acid excretion when compared with controls fed wheat bran. Because rye bran caused the most pronounced lowering effect of total cholesterol despite the lowest content of β-glucan and soluble fibers, components other than β-glucan and soluble fibers seem to be involved in its hypocholesterolemic action. Since the effects of the oat and barley ingredients were not solely correlated to the β-glucan content, structural changes occurring during processing and concentrating of the products may have impaired the hypocholesterolemic potential of the β-glucans, and other factors such as solubility and viscosity of the fiber components seem to be involved.     

Comparison of Swiss red wheat grain and fractions for their antioxidant properties

Swiss red wheat grain, bran, aleurone, and micronized aleurone were examined and compared for their free radical scavenging properties against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH*), radical cation ABTS*+ and peroxide radical anion O(2)*-, oxygen radical absorbance capacity (ORAC), chelating capacity, total phenolic content (TPC), and phenolic acid composition. The results showed that micronized aleurone, aleurone, bran, and grain may significantly differ in their antioxidant properties, TPC, and phenolic acid composition. Micronized aleurone had the greatest antioxidant activities, TPC, and concentrations of all identified phenolic acids, suggesting the potential of postharvesting treatment on antioxidant activities and availability of TPC and phenolic acids. Ferulic acid was the predominant phenolic acid in Swiss red wheat and accounted for approximately 57-77% of total phenolic acids on a weight basis. Ferulic acid concentration was well correlated with scavenging activities against radical cation and superoxide anion, TPC, and other phenolic acid concentrations, suggesting the potential use of ferulic acid as a marker of wheat antioxidants. In addition, 50% acetone and ethanol were compared for their effects on wheat ORAC values. The ORAC value of 50% acetone extracts was 3-20-fold greater than that of the ethanol extracts, indicating that 50% acetone may be a better solvent system for monitoring antioxidant properties of wheat. These data suggest the possibility to improve the antioxidant release from wheat-based food ingredients through postharvesting treatment or processing.     

In vitro digestion and fermentation characteristics of temulose molasses, a coproduct of fiberboard production, and select temulose fractions using canine fecal inoculum

It is of interest to discover new fermentable carbohydrate sources that function as prebiotics. This study evaluated the hydrolytic digestibility, fermentative capacity, and microbiota modulating properties of Temulose molasses, four hydrolyzed fractions of Temulose molasses, short-chain fructooligosaccharides (scFOS), and a yeast cell wall preparation (Safmannan). These substrates resisted in vitro hydrolytic digestion. Each substrate was fermented in vitro using dog fecal inoculum, and fermentation characteristics were quantified at 0 and 12 h. All Temulose molasses substrates decreased pH by at least 0.64 unit and resulted in greater (P < 0.05) butyrate and total short-chain fatty acid (SCFA) production compared to scFOS and Safmannan. Temulose molasses substrates resulted in higher (P < 0.01) or equal Bifidobacterium spp. concentrations compared to scFOS. Temulose molasses substrate and its fractions demonstrated prebiotic characteristics as indicated by low hydrolytic digestibility, high fermentability, and enhanced growth of microbiota considered to be beneficial to health.     

Oat Bran Fermentation by Rye Sourdough

Hydration of oat bran including fermentation by rye sourdough was studied. Three types of oat bran suspensions were prepared (a control, one with whole meal rye flour added, and one with rye starter added). The suspensions were incubated for 1, 2, 3 and 4 hr. β‐Glucan content and solubilities of protein and β‐glucan were analyzed. Viscosity of the supernatants of oat bran suspensions was determined. Neither the rye sourdough nor the rye flour alone had a significant effect on the total β‐glucan content of oat bran suspensions. However, the addition of rye, either as whole meal rye flour or as sourdough starter, markedly increased the solubility of β‐glucan and proteins and simultaneously decreased the viscosity of the water‐soluble fraction of oat bran suspension. This suggests that a hydrolysis of β‐glucan had occurred that could change the rheological properties of oat bran in baking and the physiological potential of oat bran in nutrition.     

Postprandial glycemia, insulinemia, and satiety responses in healthy subjects after whole grain rye bread made from different rye varieties. 1

Rye products typically induce low insulin responses and appear to facilitate glucose regulation. The objective of this study was to investigate differences in postprandial glucose, insulin, and satiety responses between breads made from five rye varieties. Breads made from whole grain rye (Amilo, Rekrut, Dankowski Zlote, Nikita, and Haute Loire Pop) or a white wheat bread (WWB) were tested in a randomized cross-over design in 14 healthy subjects (50 g available starch). Metabolic responses were also related to the composition of dietary fiber and bioactive compounds in the breads and to the rate of in vitro starch hydrolysis. The Amilo and Rekrut rye breads induced significantly lower insulin indices (II) than WWB. Low early postprandial glucose and insulin responses (tAUC 0-60 min) were related to higher amounts of caffeic, ferulic, sinapic, and vanillic acids in the rye breads, indicating that the phenolic acids in rye may influence glycemic regulation. All rye breads induced significantly higher subjective feelings of fullness compared to WWB. A low II was related to a higher feeling of fullness and a lower desire to eat in the late postprandial phase (180 min). The data indicate that some rye varieties may be more insulin-saving than others, possibly due to differences in dietary fiber, rate of starch hydrolysis, and bioactive components such as phenolic acids.     

Fructan Content of Rye and Rye Products

The fructan content of Finnish rye grains (13 samples, seven cultivars, harvested in 1998‐2000) varied at 4.6–6.6 g/100 g (db). Commercial whole grain rye flour and rye flakes had fructan content of 4 g/100 g, light refined rye flour had fructan content of 3 g/100 g, and rye bran had fructan content of 7 g/100 g. Fructan content as high as 23 g/100 g was detected in the water‐extractable concentrate of rye bran. Finnish soft rye bread and rye crisp bread contained 2–3 g of fructan/100 g. According to the suggested new definition of dietary fiber, fructans are also classified as dietary fiber. This means that the dietary fiber content of some cereal foods such as rye products may be increased by as much as 20% due to the presence of fructans in the grain.     

Extraction and Physicochemical Characterization of Rye β‐Glucan and Effects of Barium on Polysaccharide Molecular Weight

Use of saturated Ba(OH)₂ to extract rye β‐glucan led to a depolymerized product. Similar depolymerization of β‐glucan was observed when oat bran was extracted with this reagent. Isolated oat β‐glucan, detarium xyloglucan, guar galactomannan, and wheat and rye arabinoxylan were also depolymerized by treatment with the barium reagent. The degree of depolymerization was related to time of contact with, and concentration of, the barium. Rye β‐glucan of two different molecular weights (MW) were isolated and characterized. The structure of rye β‐glucan, as evaluated from the ratio of (1→3)‐linked cellotriosyl to (1→3)‐linked cellotetraosyl primary structural units, most closely resembles barley β‐glucan. Analytical variability of this ratio is discussed. A freshly prepared solution (2%) of the higher MW sample showed shear thinning behavior typical of cereal β‐glucans. The lower MW sample at 2% was not shear thinning, but on further purification, after storage for seven days, a 6% solution had gelled as shown by the mechanical spectrum.     

Qualitative characterization of benzoxazinoid derivatives in whole grain rye and wheat by LC-MS metabolite profiling

Benzoxazinoids are metabolites occurring in a restricted group of plant species including crops such as rye, wheat, and maize. Focus on the analysis of benzoxazinoid metabolites has typically been due to their importance to plant biochemistry and physiology as highly bioactive molecules that plants use as alleochemicals to defend themselves against predators and infections. However, the potential dietary contribution of these compounds has not been addressed. This study conducted a detailed qualitative characterization of benzoxazinoid metabolites present in the whole grain rye and processed fractions of rye bran, and their presence was also detected in whole grain wheat samples. Several novel benzoxazinoid metabolites of the hydroxamic acids (2,4-dihydroxy-1,4-benzoxazin-3-one, DIBOA; 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one, DIMBOA), lactams (2-hydroxy-1,4-benzoxazin-3-one, HBOA), and benzoxazolinones (1,3-benzoxazol-2-one, BOA) were identified, including double-hexose derivatives of DIBOA, DIMBOA, and HBOA. This paper presents an important addition to the information on the phytochemical composition of rye and wheat grains, which deserves attention in the discussion of the potential health-promoting effects of these grains.     

In Vitro Binding of Bile Acids by Rice Bran,Oat Bran,Wheat Bran,and Corn Bran

The in vitro bile acid binding by rice, oat, wheat, and corn brans was determined using a mixture of bile acids normally secreted in human bile at a physiological pH of 6.3. The objective of the study was to relate bile acid binding of cereal brans to health promoting properties. Three experiments were conducted testing substrates on an equal weight (dry matter) basis, an equal total dietary fiber (TDF) basis, and an equal TDF and equal fat basis. Each experiment was repeated to validate the results (for a total of six experiments). The relative in vitro bile acid binding of the cereal brans on an equal TDF basis considering cholestyramine as 100% bound was rice bran 51%, wheat bran 31%, oat bran 26%, and corn bran 5%. The data suggest that cholesterol lowering by rice bran appears to be related to bile acid binding. The primary mechanism of cholesterol lowering by oat bran may not be due to bile acid binding by soluble fiber. Bile acid binding did not appear to be proportional to the soluble fiber content of the cereal brans tested. Bile acid binding by wheat bran may contribute to cancer prevention and other healthful properties.     

Plant Sterols in Cereals and Cereal Products

The total plant sterol contents (free sterols and covalently bound structures) of the main cereals cultivated in Finland were determined. Furthermore, sterol contents were determined for different flour and bran fractions in the milling process of wheat and rye, as well as plant sterol contents in various milling and retail bakery products. The sample preparation procedure included acid and alkaline hydrolysis to liberate sterols from their glycosides and esters, respectively. Free sterols were extracted and, after recovery using solid‐phase extraction, derivatized to trimethylsilyl ethers for gas chromatography (GC) analysis. We used GC with a mass spectrometer (MS) for identification. When two cultivars of rye, wheat, barley, and oats grown in the same year were compared, the highest plant sterol content was observed in rye (mean content 95.5 mg/100 g, wb), whereas the total sterol contents (mg/100 g, wb) of wheat, barley, and oats were 69.0, 76.1, and 44.7, respectively. In addition, the 10 rye cultivars and breeding lines compared had total sterol contents of 70.7–85.6 mg/100 g. In the milling process of rye and wheat, the plant sterols fractionated according to the ash content of the corresponding milling product. In all cereal grain and milling product samples, sitosterol was the main sterol. The level of stanols differed in the different milling process samples; it was lower in the most refined rye and wheat flours (≈15%) than in the bran fractions (≈30% in the bran with 4% ash content). Rye bread with whole meal rye flour as the main or only ingredient was a good source of sterols. Sterol content was higher than that of wheat bread, whereas plant sterol content of other bakery products was affected by the type and amount of fat used in baking.     

In vitro fermentation of arabinoxylan oligosaccharides and low molecular mass arabinoxylans with different structural properties from wheat (Triticum aestivum L.) bran and psyllium (Plantago ovata Forsk) seed husk

Ball milling was used for producing complex arabinoxylan oligosaccharides (AXOS) and low molecular mass arabinoxylans (AX) from wheat bran, pericarp-enriched wheat bran, and psyllium seed husk. The arabinose to xylose ratio of the samples produced varied between 0.14 and 0.92, and their average degree of polymerization (avDP) ranged between 42 and 300. Their fermentation for 48 h in an in vitro system using human colon suspensions was compared to enzymatically produced wheat bran AXOS with an arabinose to xylose ratio of 0.22 and 0.34 and an avDP of 4 and 40, respectively. Degrees of AXOS fermentation ranged from 28% to 50% and were lower for the higher arabinose to xylose ratio and/or higher avDP materials. Arabinose to xylose ratios of the unfermented fractions exceeded those of their fermented counterparts, indicating that molecules less substituted with arabinose were preferably fermented. Xylanase, arabinofuranosidase, and xylosidase activities increased with incubation time. Enzyme activities in the samples containing psyllium seed husk AX or psyllium seed husk AXOS were generally higher than those in the wheat bran AXOS preparations. Fermentation gave rise to unbranched short-chain fatty acids. Concentrations of acetic, propionic, and butyric acids increased to 1.9-2.6, 1.9-2.8, and 1.3-2.0 times their initial values, respectively, after 24 h incubation. Results show that the human intestinal microbiota can at least partially use complex AXOS and low molecular mass AX. The tested materials are thus interesting physiologically active carbohydrates.     

Calcium-binding capacities of different brans under simulated gastrointestinal pH conditions. In vitro study with (45)Ca

The present study was performed to investigate calcium-binding characteristics of different brans under simulated gastrointestinal pH conditions and to explore the significance of dietary fiber, oxalate, and phytate for calcium binding. Different brans (rice, rye, soy, fine wheat, coarse wheat, and oat) and CaCl(2) solution containing (45)Ca were incubated at 37 degrees C at gastric pH (2.2) followed by buffering steps of 1 degree from pH 3.0 to pH 8.0. Total calcium binding and calcium-binding capacity of the pH 2.2 soluble bran fraction were determined. Additionally, oxalate and phytate contents of brans and solubility profiles of phytic acid were investigated. Calcium-binding capacities of brans showed a clear pH dependence. At gastric pH calcium binding was low in all brans, ranging from 0.022 to 0.040 mmol of calcium/g of bran. Soy bran, nearly phytate-free, showed higher binding values up to pH 4.0 and lower values between pH 5.0 and 8.0. In all other brans, binding values increased strongly with increasing pH in the quantitative order rice bran > coarse wheat bran > fine wheat bran > rye bran > oat bran. The solubility profiles indicate that in the cases of rye, wheat, and rice bran phytate accounts for 70-82% of their total calcium-binding capacities. The results suggest that dietary fiber makes no important contribution to calcium binding, except for soy and oat brans. Oxalate plays only a minor role in calcium binding by brans.     

Digestion residues of typical and high-beta-glucan oat flours provide substrates for in vitro fermentation

In vitro fermentabilities of the oat flour digestion residues (ODR) from two commercial oat lines with 4.7 and 5.3% beta-glucan and from two high-beta-glucan experimental lines with 7.6 and 8.1% beta-glucan were evaluated and compared with fermentations of lactulose, purified oat beta-glucan (POBG), and purified oat starch (POS). Substrates were fermented by using an in vitro batch fermentation system under anaerobic conditions for 24 h. The progress of the fermentation was studied by following the change in pH of the fermentation medium, production of short-chain fatty acids (SCFA) and gases, and consumption of carbohydrates. The substrate from the flour with the greatest amount of beta-glucan tended to have the greatest pH decline and the greatest total SCFA production. A significant correlation occurred between gas production and SCFA formation (R 2 = 0.89-0.99). Acetate was produced in the greatest amounts by all of the substrates except POBG, by which butyrate was produced in the greatest amount. More propionate and butyrate, but less acetate, were produced from high-beta-glucan ODR. With the given fermentation conditions, >80% of the total carbohydrate was depleted by the bacteria after 24 h. Glucose was the most rapidly consumed carbohydrate among other available monosaccharides in the fermentation medium. Overall, the high-beta-glucan experimental lines provided the best conditions for optimal in vitro gut fermentations.     

Steryl phenolic acid esters in cereals and their milling fractions

The steryl ferulate contents of rye and wheat grains and their milling fractions were analyzed using a reversed-phase high-performance liquid chromatographic (HPLC) method. HPLC-mass spectrometry was used for identification. In addition, steryl ferulates of some selected milling byproducts were determined. The total steryl ferulate contents of rye and wheat grains were 6.0 and 6.3 mg/100 g, respectively. Uneven distribution of steryl ferulates in the grains led to considerable differences in the milling products; their steryl ferulate contents ranged from trace amounts in flours with low ash content to 20 and 34 mg/100 g in rye and wheat brans, respectively. Campestanyl ferulate and sitostanyl ferulate were the main components, followed by campesteryl ferulate and sitosteryl ferulate, whereas sitosterol was the main component in total sterols. Among the other samples, a byproduct of rice milling (pearling dust) was the best source of steryl ferulates, its total steryl ferulate content being 119 mg/100 g, whereas no measurable amounts of steryl ferulates were measured in oat bran or pearling dust of barley. The results indicated that rye and wheat and especially their bran fractions are comparable to corn as steryl ferulate sources.