Viscosity and Solubility of β‐Glucan Extracted Under In Vitro Conditions from Barley β‐Glucan‐Fortified Bread and Evaluation of Loaf Characteristics

Fortifying bread with β‐glucan has been shown to reduce bread quality and the associated health benefits of barley β‐glucan. Fortification of bread using β‐glucan concentrates that are less soluble during bread preparation steps has not been investigated. The effects of β‐glucan concentration and gluten addition on the physicochemical properties of bread and β‐glucan solubility and viscosity were investigated using a less soluble β‐glucan concentrate, as were the effects of baking temperature and prior β‐glucan solubilization. Fortification of bread with β‐glucan decreased loaf volume and height (P ≤ 0.05) and increased firmness (P ≤ 0.05). Gluten addition to bread at the highest β‐glucan level increased height and volume (P ≤ 0.05) to values exceeding those for the control and decreased firmness (P ≤ 0.05). β‐Glucan addition increased (P ≤ 0.05) extract viscosity, as did gluten addition to the bread with the highest β‐glucan level. Baking at low temperature decreased (P ≤ 0.05) β‐glucan viscosity and solubility, as did solubilizing it prior to dough formulation. Utilization of β‐glucan that is less soluble during bread preparation may hold the key to effectively fortifying bread with β‐glucan without compromising its health benefits, although more research is required.     

Effect of Formulation and Processing Treatments on Viscosity and Solubility of Extractable Barley β‐Glucan in Bread Dough Evaluated Under In Vitro Conditions

β‐Glucan shows great potential for incorporation into bread due to its cholesterol lowering and blood glucose regulating effects, which are related to its viscosity. The effects of β‐glucan concentration, gluten addition, premixing, yeast addition, fermentation time, and inactivation of the flour enzymes on the viscosity of extractable β‐glucan following incorporation into a white bread dough were studied under physiological conditions, as well as, β‐glucan solubility in fermented and unfermented dough. β‐Glucan was extracted using an in vitro protocol designed to approximate human digestion and hot water extraction. The viscosity of extractable β‐glucan was not affected by gluten addition, the presence of yeast, or premixing. Fermentation produced lower (P ≤ 0.05) extract viscosity for the doughs with added β‐glucan, while inactivating the flour enzymes and increasing β‐glucan concentration in the absence of fermentation increased (P ≤ 0.05) viscosity. The physiological solubility of the β‐glucan concentrate (18.1%) and the β‐glucan in the unfermented dough (20.5%) were similar (P > 0.05), while fermentation substantially decreased (P ≤ 0.05) solubility to 8.7%, indicating that the reduction in viscosity due to fermentation may be highly dependent on solubility in addition to β‐glucan degradation. The results emphasize the importance of analyzing β‐glucan fortified foods under physiological conditions to identify the conditions in the dough system that decrease β‐glucan viscosity so that products with maximum functionality can be developed.     

Influence of Oat β‐Glucan Preparations on the Perception of Mouthfeel and on Rheological Properties in Beverage Prototypes

The aim was to study the effect of concentration and molecular weight of four different β‐glucan preparations on the perceived sensory quality of a beverage prototype. The correlations between sensory and instrumental measures were investigated. Two of the preparations were brantype containing high molecular weight β‐glucan, two were more‐processed low molecular weight β‐glucan preparations. Twelve beverage samples containing 0.25–2% β‐glucan and one reference sample thickened with carboxymethyl cellulose (CMC) were profiled by a sensory panel and analyzed by instrumental measurements (viscosity and molecular weight). Sensory profiles of the beverages varied at the same concentration of β‐glucan, depending on β‐glucan preparation. Beverages made with the bran‐type preparations were more viscous and had higher perceived thickness than beverages made with more‐processed, low molecular weight preparations. Moderate correlations were obtained between perceived thickness and sliminess and instrumental viscosity at all shear rates between 26 and 100/sec (r = 0.63–0.78; P ≤ 0.001). Technologically, more‐processed β‐glucan preparations are easier to add into a beverage in amounts sufficient for achieving a physiologically functional amount of β‐glucan in a product.     

Effect of Health Information on Consumer Acceptability of Bread Fortified with β‐Glucan and Effect of Fortification on Bread Quality

Fortifying bread with β‐glucan reduces bread sensorial properties, though fortification using β‐glucan concentrates of low solubility under the conditions of dough preparation has not been investigated. This study investigated the consumer acceptability and purchase intent of bread fortified with a less soluble β‐glucan concentrate at levels corresponding to 0, 0.75, and 1.5 g β‐glucan/serving bread in relation to the provision of health information, gender, and whole wheat bread consumption. The effect of β‐glucan concentration on the physical properties of the bread produced under pilot plant settings was also investigated. β‐Glucan addition decreased (P < 0.05) loaf volume, increased firmness, and produced a darker, redder bread (P < 0.05), though fortification at 1.5 g β‐glucan/serving bread decreased height as well (P < 0.05). Consumer evaluation (n = 122) revealed that health information increased liking of appearance, flavor, and overall acceptability of the 1.5 g/serving bread to levels similar to or exceeding that of the control. Liking of the 1.5 g β‐glucan/serving bread appearance increased more in women than in men and for consumers who regularly consumed whole wheat bread for perceived health benefits when β‐glucan health information was provided. The provision of β‐glucan health information may be the key to increasing consumer acceptability of bread fortified with β‐glucan.     

Effect of Processing on Viscosity and Molecular Weight of (1,3)(1,4)‐β‐Glucan in Western Australian Oat Cultivars

Six Australian milling oat cultivars grown over two growing seasons were characterized for differences in (1,3)(1,4)‐β‐glucan (β‐glucan) viscosity, solubility, molecular weight (M_w), and the effect of processing. Oat cultivars grown in 2012 had significantly higher extracted β‐glucan viscosity from oat flour than the same oat cultivar grown in 2011 (P < 0.05, mean 137 and 165 cP, respectively). Noodle β‐glucan mean viscosity for 2012 (147 cP) was significantly higher than for 2011 (128 cP). β‐Glucan from ‘Williams’ and ‘Mitika’ oats had the highest viscosity (P < 0.05) in flour (5.92 and 5.25%, respectively) and noodles (1.64 and 1.47%, respectively) for both years, compared with the other oat cultivars. β‐Glucan (M_w) of Williams for 2012 and ‘Kojonup’ for both years were the least affected by processing, with an average drop of 33% compared with a maximum of 63% for other cultivars. Therefore, Williams showed superior β‐glucan properties to other oat cultivars studied, and can potentially provide improved health benefits. High and low β‐glucan M_w populations were found in the same elution peak after processing. Oat cultivars chosen for processing should be those with β‐glucans that are more resistant to processing, and that maintain their physiochemical properties and, therefore, bioactivity.     

Modification of Standard Enzymatic Protocol to a Cost‐Efficient Format for Mixed‐Linkage (1→3,1→4)‐β‐d‐Glucan Measurement

The current enzymatic assay approach (AACC International Approved Method 32‐23) for the measurement of mixed‐linkage β‐glucan in small grains was modified to a cost‐efficient and high‐throughput format without compromising the accuracy of the results. Ten barley (Hordeum vulgare L.) genotypes used in the study represented a wide range of β‐glucan content levels. A reduced reaction volume is used in the new protocol to adapt to a 96‐well plate format. The volume of key components lichenase and β‐glucosidase were reduced to 25% of the volume required in the original protocol and the cost per sample was reduced to 22% of that in the original protocol. Labor cost was also decreased to 25% of the original protocol as a result of format changes. The accuracy of the measurement from the modified protocol was comparable to the current standard enzymatic procedure. β‐Glucan measurement accuracy of the modified and original protocols were also compared using 21 oat (Avena sativa L.) samples. The results indicated that the new protocol consistently produced accurate measurements in both barley and oat.     

Comparison of Different Types of NIR Instruments in Ability to Measure β‐Glucan Content in Naked Barley

Importance of β‐glucan in human nutrition is mirrored in numerous approval applications registering β‐glucan containing products as health beneficial products in accordance with forthcoming EU Health Claims Regulation. In comparison to other cereals, barley contains considerable amounts of β‐glucan. Naked barley is of particular interest because it circumvents the costs and loss of beneficial substances related to dehusking. In this study, the potential of near‐infrared spectroscopy as an accurate, fast and economic method of determination of β‐glucan in naked barley was appraised. Four different near‐infrared instruments were used to analyze 107 barley samples, in both whole grain and milled form. Importantly, both black and purple pericarp samples, which are of additional nutritional interest due to high anthocyanin content, and waxy samples, which show an extraordinary high β‐glucan content could be analyzed within the same calibration set as the normal samples. All tested dispersive near‐infrared reflection instruments showed suitability for supervision of breeding experiments and β‐glucan monitoring in food industries (R² > 0.78). Common, industrially used near‐infrared transmission instruments also provided reasonable results, although only suitable for rough selection according to β‐glucan levels. On the other hand, the Fourier transform near‐infrared reflection instrument was able to perform analytical analyses (R² = 0.96–0.98).     

REVIEW: Oat and Rye β‐Glucan: Properties and Function

Over the years, the β‐glucan of oats and barley has been the subject of study either because of the importance of the cholesterol‐lowering potential to health claims (FDA 1997, 2005) or, in the case of barley, because of the role of β‐glucan and β‐glucan‐rich endosperm cell walls in malting and brewing. β‐Glucan is also present in rye and in much lesser amounts in wheat. The most striking difference in these latter two sources is the difficulty in extractability; alkali rather than water is required for significant release from the cell walls. This review will discuss physicochemical properties of oat and rye β‐glucan and, where information allows, relate these to physiological effects. Viscosity, or more generally rheology, plays a central role in discussions of cereal β‐glucan functionality and physiological effects and will be the focus of this review.     

Factors Influencing β‐Glucan Levels and Molecular Weight in Cereal‐Based Products

The beneficial role of soluble dietary fiber in human nutrition is well documented and has lead to a growing demand for the incorporation of β‐glucan, particularly from oats and barley, into foods. β‐Glucan with high solubility and high molecular weight distribution results in increased viscosity in the human intestine, which is desirable for increased physiological activity. Molecular weight, level, and solubility of β‐glucan are affected by genotype, environment, agronomic input, and the interactions of these factors and food processing methods. Available literature reveals that the level of β‐glucan in a finished product (e.g. bread, cake, muffins) depends upon several factors in the production chain, whereas food processing operations are major factors affecting molecular weight and solubility of β‐glucans. Therefore, to avail themselves of the natural bioactive compounds, food manufacturers must pay attention not only to ensure sufficient concentration of β‐glucan in the raw material but also to the processing methods and functional properties of β‐glucan, minimizing enzymatic or mechanical breakdown of the β‐glucans in end‐product and optimizing processing conditions. This review discusses the different sources of β‐glucan for use in human functional foods and factors affecting the levels and the molecular weight of β‐glucan at various pre‐ and postharvest operations.     

Effects of β‐Glucan Content and Pearling of Barley in Diet‐Induced Obese Mice

The effects of the β‐glucan content and pearling of barley on abdominal obesity and the proinflammatory state were investigated in diet‐induced obese mice. Male C57BL/6J mice were randomly divided into four groups and fed either a high‐fat diet containing high‐β‐glucan barley (Beau Fiber [BF]) or a high‐fat diet containing β‐glucan‐free barley (Shikoku‐hadaka 84(bgl ) [BGL]) as whole grain flour or 60% pearled flour for 12 weeks. The weights of mesenteric fat, serum total and low density lipoprotein cholesterol levels, serum insulin and fasting glucose levels, oral glucose tolerance test results, and messenger RNA (mRNA) expression of proinflammatory markers in epididymal fat in both BF groups were significantly lower than those of both BGL groups. The abundance of Bacteroides in both BF groups was significantly higher than that in both BGL groups, whereas the abundance of Clostridium clusters in both BF groups was significantly lower than that in both BGL groups. No significant differences between the whole grain and pearled flours were observed. These results suggest that high‐β‐glucan barley attenuates the progression of abdominal obesity and the proinflammatory state in diet‐induced obese mice compared with β‐glucan‐free barley, possibly by modifying insulin secretion and the microbiota.     

Distribution of β‐Glucan in the Grain of Hull‐less Barley

Nine hull‐less barley (HB) containing waxy (0–7% amylose), normal (≈25% amylose), or high amylose (≈42% amylose) starch with normal or fractured granule make‐up and 4–9% (1→3)(1→4)‐β‐d ‐glucans (β‐glucan) were pearled to remove 70% of the original grain weight in 10% intervals. The pearled fractions were analyzed for β‐glucan distribution within HB grain. Protein content of the pearled fractions indicated that the three outermost fractions contained pericarp and testa, aleurone, and subaleurone tissues, respectively. For all HB, β‐glucan and acid‐extract viscosity were very low in the outermost 20% of the kernel. For low β‐glucan HB, β‐glucan content was the greatest in the subaleurone region and declined slightly toward inner layers. For high β‐glucan HB, however, more than 80% of grain β‐glucan was distributed more evenly throughout the endosperm. Acid extract viscosity was significantly (P < 0.01) correlated with total (r = 0.75) and soluble (r = 0.87) β‐glucan content throughout the kernel of all HB. Growing conditions, location and year, had significant effects on the concentration of protein, starch and β‐glucan. However, protein, starch, and β‐glucan distribution patterns were not affected by growing conditions. The difference in β‐glucan distribution between low and high β‐glucan HB may explain the difference in milling performance of HB with low or high β‐glucan.     

Isolation,Fractionation, and Structural Characteristics of Alkali‐Extractable β‐Glucan from Rye Whole Meal

The main nonstarch polysaccharide of rye is arabinoxylan (AX), but rye contains significant levels of (1→3)(1→4)‐β‐d ‐glucan, which unlike oat and barley β‐glucan, is not readily extracted by water, possibly because of entrapment within a matrix of AX cross‐linked by phenolics. This study continues objectives to improve understanding of factors controlling the physicochemical behavior of the cereal β‐glucans. Rye β‐glucan was extracted by 1.0N NaOH and increasing concentrations of ammonium sulfate were used to separate the β‐glucan from AX and prepare a series of eight narrow molecular weight (MW) distribution fractions. Composition and structural characteristics of the isolated β‐glucan and the eight fractions were determined. High‐performance size‐exclusion chromatography (HPSEC) with both specific calcofluor binding and a triple detection (light scattering, viscometry, and refractive index) system was used for MW determination. Lichenase digestion followed by high‐performance anion exchange chromatography of released oligosaccharides, was used for structural evaluation. The overall structure of all fractions was similar to that of barley β‐glucan.     

Glycemic Response to Oat Bran Muffins Treated to Vary Molecular Weight of β‐Glucan

Oat bran muffins, containing 4 or 8 g of β‐glucan per two‐muffin serving, were prepared with or without β‐glucanase treatment to produce a range of β‐glucan molecular weights from 130,000 to just over 2 million. Following an overnight fast, the glycemic responses elicited by the untreated and treated muffins was measured in 10 healthy subjects and compared with a control whole wheat muffin. Taken all together, the 4‐g β‐glucan/serving muffins reduced blood glucose peak rise (PBGR) by 15 ± 6% compared with the control. The 8‐g β‐glucan/serving muffins had a significantly greater effect (44 ± 5% reduction compared with the control, P < 0.05). The efficacy of the muffins decreased as the molecular weight was reduced from a 45 ± 6% reduction in PBGR (P < 0.05) for the untreated muffins (averaged of both serving sizes) to 15 ± 6% (P < 0.05) for muffins with the lowest molecular weight. As the molecular weight was reduced from 2,200,000 to 400,000, the solubility of the β‐glucan increased from a mean of 44 to 57%, but as the molecular weight was further decreased to 120,000, solubility fell to 26%. There was a significant correlation (r² = 0.729, P < 0.001) between the peak blood glucose and the product of the extractable β‐glucan content and the molecular weight of the β‐glucan extracted.     

Release of β‐Glucan from Cell Walls of Starchy Endosperm of Barley

β‐Glucan can be solubilized from barley by warm water, with increasing solubilization as the temperature is increased. Substantially less glucan is extracted if the barley is dehusked using sulfuric acid, particularly if the dehusked barley is denatured. This indicates that enzymes capable of solubilizing glucan are present in barley. Various purified enzymes promote the solubilization of glucan from denatured and dehusked barley. Apart from endo‐β‐(1→3)(1→4)‐glucanase, these enzymes include endo‐xylanases, arabinofuranosidase, xyloacetylesterase, and feruloyl esterase. Ferulic acid and, probably, acetyl groups are esterlinked to arabinoxylan, not β‐glucan, in the cell walls of barley starchy endosperm, so the ability of the esterases, xylanases, and arabinofuranosidase to solubilize glucan indicates the pentosan component of the cell wall can restrict the extraction of glucan.     

Gastrointestinal Release of β‐Glucan and Pectin Using an In Vitro Method

The release of soluble dietary fiber is a prerequisite for viscous effects and hence beneficial health properties. A simple in vitro method was adapted to follow the release during gastrointestinal digestion, and the percentage of solubilized fiber was measured over time. β‐Glucan from oat bran was mainly released during gastric digestion while the release of pectin from sugar beet fiber continued in the small intestine. Unmilled fractions of sugar beet fiber released more soluble fiber than oat bran flakes, probably due to the porous structure of sugar beet fiber as a result of manufacturing processes, but also due to differences in source. Milling to smaller fiber particles significantly improved releasability (from 20 to 55% released β‐glucan and from 50 to 70% released pectin, respectively, after digestion). When milled fibers were included in individual food matrices, the release was reduced by protein and starch matrices (5% β‐glucan and 35% pectin released, respectively) and slowed by fat (45% β‐glucan and 60% pectin released). This may result in a too low or too late release in the upper small intestine to be able to interfere with macronutrient uptake. The method may be suitable for predicting the gastrointestinal release of soluble dietary fibers from food matrices in the development of healthy food products.     

Bulk Carbohydrate Grain Filling of Barley β‐Glucan Mutants Studied by 1H HR MAS NMR

Temporal and genotypic differences in bulk carbohydrate accumulation in three barley genotypes differing in the content of mixed linkage β‐(1→3),(1→4)‐D‐glucan (β‐glucan) and starch were investigated using proton high‐resolution, magic angle spinning, nuclear magnetic resonance (¹H HR MAS NMR) during grain filling. For the first time, ¹H HR MAS NMR spectra of flour from immature barley seeds are analyzed. Spectral assignments are made using two‐dimensional (2D) NMR methods. Both α‐ and β‐glucan biosynthesis were characterized by inspection of the spectra as well as by calibration to the reference methods for starch and β‐glucan content. Starch was quantified with very good calibrations to the α‐(1→4) peak (5.29–5.40 ppm) and the region 3.67–3.83 ppm covering starch glycopyranosidic protons from H5 and H6. In contrast, the spectral inspection of the β‐anomeric region 4.45–4.85 ppm showed unexpected lack of intensity in the high β‐glucan mutant lys5f at seed maturity, resulting in poor calibration to reference β‐glucan content. We hypothesize that the lack of β‐glucan signal in lys5f indicates partial immobilization of the β‐glucan that appears to be either genotypic dependent or water/β‐glucan ratio dependent.     

Prediction of β‐Glucan Concentration Based on Viscosity Evaluations of Raw Oat Flours from High β‐Glucan and Traditional Oat Lines

Rheological properties of raw oat flour slurries were determined in experimental high β‐glucan (≤7.8%) and traditional oat lines (4–5% β‐glucan) grown in two consecutive years. Three different media were used to disperse oat flours: deionized water, silver nitrate solution (to inactivate endogenous enzymes), and alkali solution (to solubilize both water‐soluble and water‐insoluble β‐glucans). Significant correlations (P < 0.05) between viscosity of slurries and β‐glucan concentration obtained in either deionized water (r = 0.833), silver nitrate (r = 0.940), or alkali (r = 0.896) solutions showed that β‐glucans were the main contributor to oat extract viscosity. The highest correlation was obtained in silver nitrate solution, suggesting that inactivating endogenous enzymes is important to obtain high correlations. Predictive models of oat β‐glucan concentration based on the viscosity profile were developed using partial least squares (PLS) regression. Prediction of β‐glucan concentration based on viscosity was most effective in the silver nitrate solution (r = 0.949, correlation coefficient of predicted vs. analyzed β‐glucans) and least effective in the alkali solution (r = 0.870). These findings demonstrate that the β‐glucan in oat could be predicted by measuring the viscosity of raw flours in silver nitrate solution, and this method could be used as a screening tool for selective breeding.     

Pasting and Thermal Properties of Flours from Oat Lines with High and Typical Amounts of β‐Glucan

Experimental oat lines high in β‐glucan (6–7.8%) and traditional lines (3.9–5.7% β‐glucan) were used to evaluate the effect of β‐glucan on pasting (by rapid viscoanalysis) and thermal properties (by differential scanning calorimetry) of oat flours. Significant correlations established between β‐glucan concentration and the pasting parameters after amylolysis demonstrated the role of β‐glucans in oat pasting. The relative decrease of peak viscosity (PV) observed after enzymatic removal of β‐glucans was correlated with β‐glucan concentration (r = 0.880, P < 0.010) and reconfirmed their contribution to pasting. A significant increase of PV with β‐glucan concentration obtained under conditions of either autolysis (deionized water used for dispersion) (r = 0.89, P < 0.010) or inhibition (silver nitrate solution used for dispersion) (r = 0.91, P < 0.001) might be explained by an increase in water retention capacity caused by the β‐glucans. Predictive models of β‐glucan concentration based on the whole pasting profile or selected profile regions were developed using partial least squares (PLS) regression.Prediction of β‐glucan based on the whole profile obtained in the silver nitrate solution was the most effective (r = 0.93, correlation coefficient of predicted vs. analyzed β‐glucans, P < 0.050). No correlations were observed between the thermal properties of oat flours and the β‐glucan concentration.     

Structure of (1→3)(1→4)‐β‐d‐Glucan in Waxy and Nonwaxy Barley

The endosperm cell walls of barley are composed largely of a (1→3)(1→4)‐β‐d ‐glucan commonly known simply as β‐d ‐glucan (Wood 2001). There has been much research into the characteristics of barley β‐glucan because of the influence of this polysaccharide on performance of barley in malting and subsequent brewing of beer, and in feed value, especially for young chicks (MacGregor and Fincher 1993). The potential for β‐glucan to develop high viscosity is a problem in these uses, but from the perspective of human nutrition, this characteristic may be an advantage. The glycemic response to oat β‐glucan is inversely related to (log)viscosity (Wood et al 1994a) and there is evidence to suggest that the lowering of serum cholesterol levels associated with oat and barley products (Lupton et al 1994; Wood and Beer 1998) is at least in part due to the β‐glucan (Braaten et al 1994) and probably also its capacity to develop viscosity in the gastrointestinal tract (Haskell et al 1992).     

Preparation and Characterization of Films Using Barley and Oat β‐Glucan Extracts

Films for potential food use were prepared from aqueous solutions of β‐glucan extracted from hulled barley, hull‐less barley, and oats. The extracts (75.2–79.3% β‐glucan) also contained proteins, fat, and ash. Glycerol was used as a plasticizer. The films were translucent, smooth, and homogeneous in structure on both sides. Water vapor permeability of films prepared from 4% solutions of β‐glucan extracts were higher than those from 2% solutions, despite similar values for water vapor transmission rate. Mechanical properties were influenced by both β‐glucan source and concentration. The oat β‐glucan films showed higher tensile strength and water solubility, and lower color, opacity, and deformation values than those of barley. Films prepared from hull‐less barley cv. HLB233 remained intact upon immersion in water for 24 hr.