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β-Glucan Assay Kit (Mixed Linkage)

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00:03    Introduction
00:48    Principle
01:37    Reagent Preparation
04:50    Weighing of samples
06:20    Gelatinisation of sample
07:08    Lichenase depolymerisation of β-Glucan
08:30    pH adjustment & incubation with β-Glucosidase
10:30    Glucose Determination (GOPOD Reagent)
12:04    Calculations
14:30    Further information

beta-Glucan Assay Kit Mixed Linkage K-BGLU Scheme
Product code: K-BGLU

100 assays per kit

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Content: 100 assays per kit
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 2 years under recommended storage conditions
Analyte: β-Glucan
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 510
Signal Response: Increase
Linear Range: 4 to 100 μg of D-glucose per assay
Limit of Detection: 0.5 g/100 g
Total Assay Time: ~ 100 min
Application examples: Oats, barley, malt, wort, beer, food and other materials.
Method recognition: AACC Method 32-23.01, AOAC Method 995.16, AOAC Method 992.28, CODEX Method Type II, EBC Method 3.10.1, ICC Standard No. 166 and RACI Standard Method

The Beta-Glucan test kit is suitable for the measurement and analysis of Beta-Glucan (Mixed Linkage).

For the measurement of 1,3:1,4-β-D-glucan in cereal grains, milling fractions, wort, beer and other food products.

See our complete range of polysaccharide assay kits.

Scheme-K-BGLU BGLU Megazyme

  • Very cost effective 
  • All reagents stable for > 2 years as supplied 
  • Only enzymatic kit available 
  • Very specific 
  • Simple format 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Standard included
Validation of Methods
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Other automated assay procedures Product Performance Validation Report
Megazyme publication
Measurement of carbohydrates in grain, feed and food.

McCleary, B. V., Charnock, S. J., Rossiter, P. C., O’Shea, M. F., Power, A. M. & Lloyd, R. M. (2006). Journal of the Science of Food and Agriculture, 86(11), 1648-1661.

Procedures for the measurement of starch, starch damage (gelatinised starch), resistant starch and the amylose/amylopectin content of starch, β-glucan, fructan, glucomannan and galactosyl-sucrose oligosaccharides (raffinose, stachyose and verbascose) in plant material, animal feeds and foods are described. Most of these methods have been successfully subjected to interlaboratory evaluation. All methods are based on the use of enzymes either purified by conventional chromatography or produced using molecular biology techniques. Such methods allow specific, accurate and reliable quantification of a particular component. Problems in calculating the actual weight of galactosyl-sucrose oligosaccharides in test samples are discussed in detail.

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Megazyme publication

Determination of β-glucan in barley and oats by streamlined enzymic method: summary of collaborative study.

McCleary, B. V. & Mugford, D. C. (1997). Journal of AOAC International, 80(3), 580-583.

A collaborative study was conducted involving 8 laboratories (including the authors’ laboratories) to validate the streamlined enzymatic method for determination of β-D-glucan in barley and oats. In the method, the flour sample is cooked to hydrate and gelatinize β-glucan, which is subsequently hydrolyzed to soluble fragments with the lichenase enzyme. After volume and pH adjustments and filtration, the solution is treated with β-glucosidase, which hydrolyzes β-gluco-oligosaccharides to D-Glucose. D-Glucose is measured with glucose oxidase–peroxidase reagent. Other portions of lichenase hydrolysate are treated directly with glucose oxidase-peroxidase reagent to measure free glucose in test sample. If levels of free glucose are high, the sample is extracted first with 80% ethanol. For all samples analyzed, the repeatability relative standard deviation (RSDr) values ranged from 3.1 to 12.3% and the reproducibility relative standard deviation (RSDr) values ranged from 6.6 to 12.3%. The streamlined enzymatic method for determination of β-D-glucan in barley and oats has been adopted first action by the AOAC INTERNATIONAL.

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Megazyme publication
Measurement of total starch in cereal products by amyloglucosidase-alpha-amylase method: collaborative study.

McCleary, B. V., Gibson, T. S. & Mugford, D. C. (1997). Journal of AOAC International, 80, 571-579.

An American Association of Cereal Chemists/AOAC collaborative study was conducted to evaluate the accuracy and reliability of an enzyme assay kit procedure for measurement of total starch in a range of cereal grains and products. The flour sample is incubated at 95 degrees C with thermostable alpha-amylase to catalyze the hydrolysis of starch to maltodextrins, the pH of the slurry is adjusted, and the slurry is treated with a highly purified amyloglucosidase to quantitatively hydrolyze the dextrins to glucose. Glucose is measured with glucose oxidase-peroxidase reagent. Thirty-two collaborators were sent 16 homogeneous test samples as 8 blind duplicates. These samples included chicken feed pellets, white bread, green peas, high-amylose maize starch, white wheat flour, wheat starch, oat bran, and spaghetti. All samples were analyzed by the standard procedure as detailed above; 4 samples (high-amylose maize starch and wheat starch) were also analyzed by a method that requires the samples to be cooked first in dimethyl sulfoxide (DMSO). Relative standard deviations for repeatability (RSD(r)) ranged from 2.1 to 3.9%, and relative standard deviations for reproducibility (RSD(R)) ranged from 2.9 to 5.7%. The RSD(R) value for high amylose maize starch analyzed by the standard (non-DMSO) procedure was 5.7%; the value was reduced to 2.9% when the DMSO procedure was used, and the determined starch values increased from 86.9 to 97.2%.

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Megazyme publication
Measurement of (1→3),(1→4)-β-D-glucan in barley and oats: A streamlined enzymic procedure.

McCleary, B. V. & Codd, R. (1991). Journal of the Science of Food and Agriculture, 55(2), 303-312.

A commercially available enzymic method for the quantitative measurement of (1→3),(1→4)-β-glucan has been simplified to allow analysis of up to 10 grain samples in 70 min or of 100–200 samples by a single operator in a day. These improvements have been achieved with no loss in accuracy or precision and with an increase in reliability. The glucose oxidase/peroxidase reagent has been significantly improved to ensure colour stability for periods of up to 1 h after development. Some problems experienced with the original method have been addressed and resolved, and further experiments to demonstrate the quantitative nature of the assay have been designed and performed.

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Megazyme publication
Purification of (1→3),(1→4)-β-D-glucan from Barley Flour.

McCleary, B. V. (1988). “Methods in Enzymology”, Volume 160, (H. Gilbert, Ed.), Elsevier Inc., pp. 511-514.

The major endosperm cell wall polysaccharide in barley and oats is a linear (1→3),(1→4)-β-D-glucan. In barley, it represents approximately 75% of the carbohydrate in endosperm cell walls. It is generally considered that the majority of the polysaccharide consists of two or three 1,4-β-linked D-glucosyl residues, joined by single 1,3-β-linkages. Barley flour contains mixed-linkage β-glucan fractions that vary in their ease of extraction. Methods employed for the extraction and purification of barley β-glucan are generally modifications of the procedure described by Preece and Mackenzie. Extraction efficiency may be related to the time and conditions of storage of the grain or flour or to conditions of pretreatment of the grain before extraction. Exhaustive extraction procedures have been applied to isolated endosperm cell wall preparations that are essentially devoid of starch and protein. This chapter describes a modification of the method of Preece and Mackenzie that allows the large scale, essentially quantitative extraction of mixed-linkage β-glucan from barley flour.

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Megazyme publication
Measurement of (1→3),(1→4)-β-D-glucan.

McCleary, B. V., Shameer, I. & Glennie-Holmes, M. (1988). Methods in Enzymology, 160, 545-551.

The major carbohydrate component of the endosperm cell walls of barley and oat grain is a mixed-linkage (1→3),(1→4)-β-D-glucan commonly termed barley β-glucan. Barley β-glucan forms highly viscous aqueous solutions and gelatinous suspensions. In the brewing industry it can lead to diminished rates of wort and beer filtration and to the formation of hazes, precipitates, and gels in stored beer. In an attempt to alleviate the problems caused by barley β-glucan in the brewing and animal feed industries, various approaches have been adopted including the breeding of barley varieties low in this component, the use of only well-modified malts in brewing, and the addition of enzymes active on barley β-glucan. None of these methods has been adopted as a standard procedure. Reasons for this include the lack of specificity or reliability of the assay or the tedious nature of the assay format that limits the number of samples that can be processed in a given time. This chapter describes an assay procedure that overcomes these limitations. In this assay, highly purified endo-1,3(4)-β-glucanase (lichenase) and β-glucosidase are employed. The glucan is depolymerized by lichenase to oligosaccharides, these oligosaccharides are quantitatively hydrolyzed by β-glucosidase to glucose, and this is specifically measured using glucose oxidase/peroxidase reagent. This method is suitable for the routine analysis of mixed-linkage β-glucan in cereal flours, malt, wort, and beer.

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Megazyme publication
Measurement of (1→3)(1→4)-β-D-glucan in malt, wort and beer.

McCleary, B. V. & Nurthen, E. (1986). Journal of the Institute of Brewing, 92(2), 168-173.

A method developed for the quantification of (1→3)(1→4)-β-D-glucan in barley flour has been modified to allow its use in the measurement of this component in malt, wort, beer and spent grain. For malt samples, free D-glucose was first removed with aqueous ethanol. Quantification of the polymer in wort and beer samples involved precipitation of the β-glucan with ammonium sulphate followed by washing with aqueous ethanol to remove free D-glucose. Spent grain was lyophilised and milled and then analysed by the method developed for malt. In all cases, the β-glucan was depolymerised with lichenase and the resultant β-gluco-oligosaccharides hydrolysed to D-glucose with β-D-glucosidase. The released D-glucose was then specifically determined using glucose oxidase-peroxidase reagent.

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Megazyme publication
Enzymic hydrolysis and industrial importance of barley β-glucans and wheat flour pentosans.

McCleary, B. V., Gibson, T. S., Allen, H. & Gams, T. C. (1986). Starch, 38(12), 433-437.

Mixed linkage β-glucane and pentosanes (mainly arabinoxylanes) are the major endosperm cell-wall polysaccharides of barley and wheat respectively. These polysaccharides, although minor components of the whole grain, significantly affect the industrial utilization of these cereals. The modification of barley corns during malting requires the dissolution of the β-glucan in the cell-wall of the starch endosperm. High β-glucane concentration in wort and beer effect the rate of filtration and can also lead to precipitate or gel formation in the final product. In a similar manner, pentosane is thought to cause filtration problems with wheat starch hydrolysates by increasing viscosity and by producing gelatinous precipitate which blocks filters. Ironically, it is this same viscosity building and water binding capacity which is considered to render pentosanes of considerable value in dough development and bread storage (anti-staling functions). In the current paper, some aspects of the beneficial and detrimental effects of pentosans and β-glucan in the industrial utilization of wheat and barley are discussed. More specifically, enzymic methods for the preparation, analysis and identification of these polysaccharides and for the removal of their functional properties, are described in detail.

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Megazyme publication
Enzymic quantification of (1→3) (1→4)-β-D-glucan in barley and malt.

McCleary, B. V. & Glennie-Holmes, M. (1985). Journal of the Institute of Brewing, 91(5), 285-295.

A simple and quantitative method for the determination of (1→3) (1→4)-β-D-glucan in barley flour and malt is described. The method allows direct analysis of β-glucan in flour and malt slurries. Mixed-linkage β-glucan is specifically depolymerized with a highly purified (1→3) (1→4)-β-D-glucanase (lichenase), from Bacillus subtilis, to tri-, tetra- and higher degree of polymerization (d.p.) oligosaccharides. These oligosaccharides are then specifically and quantitatively hydrolysed to glucose using purified β-D-glucosidase. The glucose is then specifically determined using glucose oxidase/peroxidase reagent. Since barley flours contain only low levels of glucose, and maltosaccharides do not interfere with the assay, removal of low d.p. sugars is not necessary. Blank values are determined for each sample allowing the direct measurement of β-glucan in values are determined for each sample allowing the direct measurement of β-glucan in malt samples. α-Amylase does not interfere with the assay. The method is suitable for the routine analysis of β-glucan in barley samples derived from breeding programs; 50 samples can be analysed by a single operator in a day. Evaluation of the technique on different days has indicated a mean standard error of 0-1 for barley flour samples containing 3-8 and 4-6% (w/w) β-glucan content.

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Megazyme publication

Enzymic modification and quantification of polymers based on a (1→4)-β-D-glucan backbone.

McCleary, B. V. (1985). “Gums and Stabilisers for the Food Industry”, Volume 3, (G. O. Philips, D. J. Wedlock and P. A. Williams, Eds.), Pergamon Press, pp. 17-28.

In this paper, examples of the use of enzymes in the modification, quantification and investigation of fine-structural details of mixed-linkage (1+3)(1+4)-β-D-glucans, xyloglucans, glucomannans and xanthan are presented and discussed.

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Molecular conformation and dilute solution properties of barley β-glucan: unveiling β-glucan as a highly flexible biopolymer under different processing conditions. 

Hematian Sourki, A. & Hesarinejad, M. A. (2023). Chemical and Biological Technologies in Agriculture, 10(1), 69.

Background: The functional properties of food fluids containing hydrocolloids are influenced by temperature, soluble salts, pH and the presence of sugars. In this research, the properties of a dilute barley β-glucan (BBG) solution were evaluated in the presence of various factors. Different models were explored to determine the intrinsic viscosity of BBG. Results: The results indicated that the models of Higiro and Tanglertpaibul-Rao were more efficient in determining the intrinsic viscosity of BBG at different temperatures, different pH, the presence of sodium chloride, calcium chloride, and sucrose. Every 10°C increase in temperature from 25 to 65°C caused a decrease in intrinsic viscosity by 7.8, 10, 5.7 and 7.2%, respectively. In relation to the non-ionic structure of BBG, the presence of monovalent and bivalent salts had a negligible effect on reducing the intrinsic viscosity. The increase in pH from 3 to 7 caused a rise in intrinsic viscosity. But a further increase in pH, up to 9, caused a decrease in intrinsic viscosity. However, these changes were not significant and indicated that the non-ionic structure of BBG was independent of the pH. Since the constant b values of BBG were close to 1 at all temperatures, salt concentrations, different pH values and different sucrose concentrations, it can be assumed that the structure of BBG in the dilute range was close to the random coil conformation. Conclusions: The values of flexibility index and activation energy for BBG were calculated as 789.52 and 0.65×107 J/kmole, respectively, which indicated that this hydrocolloid was highly flexible in different environmental conditions and it was independent of the processing temperature. Therefore, BBG can be recommended as a natural thickener in food-related fluids.

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High-beta-glucan and low-glycemic index functional bulgur produced from high-beta-glucan barley.

Tekin-Cakmak, Z. H., Ozer, C., Ozkan, K., Yildirim, H., Sestili, F., Jilal, A., Sagdic, O., Ozgolet, M. & Koksel, H. (2024). Journal of Functional Foods112, 105939.

A high β-glucan hull-less barley (cv. Chifaa) was used in bulgur production and its technological and nutritional properties were compared with bulgurs of another hull-less barley and durum wheat. Although Chifaa bulgur had longer cooking time (9.5 min) which is expected to have an increasing effect on total organic matter (TOM), it had lower TOM (1.31 g/100 g) than the durum bulgur. The phenolics of barleys were significantly higher than those of Kiziltan wheat. The amounts of total phenolic contents decreased after bulgur production. While glycemic index (GI) of durum bulgur was high, GI of barley bulgurs were medium probably due to their higher β-glucan contents. The GI of Chifaa (56.25) is very close to the limit value of low GI foods (56). The limit to bear the health claim is 3 g of β-glucans/serving. The results indicated that this can be provided per serving of high β-glucan barley bulgur.

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Effect of non-starch components on the structural properties, physicochemical properties and in vitro digestibility of waxy highland barley starch.

Xie, J., Cheng, L., Li, Z., Li, C., Hong, Y. & Gu, Z. (2024). International Journal of Biological Macromolecules, 255, 128013.

Highland barley (HB) endosperm with an amylose content of 0-10% is called waxy HB (WHB). WHB is a naturally slow-digesting grain, and the interaction between its endogenous non-starch composition and the WHB starch (WHBS) has an important effect on starch digestion. This paper focuses on the mechanisms by which the components of β-glucan, proteins and lipids affect the molecular, granular, crystalline structure and digestive properties of WHBS. After eliminating the main nutrients except for starch, the estimated glycemic index (eGI) of the samples rose from 62.56% to 92.93%, and the rapidly digested starch content increased from 60.81% to 98.56%, respectively. The resistant starch (RS) content, in contrast, dropped from 38.61% to 0.13%. Comparatively to lipids, β-glucan and protein contributed more to the rise in eGI and decline in RS content. The crystalline characteristics of starch were enhanced in the decomposed samples. The samples' gelatinization properties improved, as did the order of the starch molecules. Protein and β-glucan form a dense matrix on the surface of WHBS particles to inhibit WHBS digestion. In summary, this study revealed the mechanism influencing the digestibility of WHBS from the perspective of endogenous non-starch composition and provided a theoretical basis to develop slow-digesting foods.

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Changes in bioactive compounds during solid‐state fermentation of Sphenarium purpurascens with Aspergillus oryzae.

Pérez‐Rodríguez, E., Reyes‐Herrera, A., Ibarra‐Herrera, C. C. & Pérez‐Carrillo, E. (2023). International Journal of Food Science & Technology, 58(12), 6653-6659.

Solid-state fermentation (SSF) can improve the availability of nutrients in food products by converting complex molecules into simpler ones with the help of microorganisms. The present research focuses on using the fungi Aspergillus oryzae in SSF to ferment grasshoppers (Sphenarium purpurascens). The fermentation was carried out for 8 days, and kinetic growth, enzyme, and bioactive compound (TPC, ABTS, and DPPH) production were monitored. Second order equation modelled the growth of A. oryzae on a grasshopper substrate. During fermentation, a protein increase of 50% was obtained after day 4. TPC, ABTS, DPPH scavenging activity percentage, β-glucan, and amylase activity increased 116%, 70%, 10%, 380%, and 629.6% at 8, 0.5, 3, 4, and 7 days, respectively, as maximum during fermentation. Results herein clearly indicate that A. oryzae could use S. purpurascens as a substrate in SSF to produce enzymes and bioactive compounds of industrial and nutritional interest.

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Maiorca wheat malt: A comprehensive analysis of physicochemical properties, volatile compounds, and sensory evaluation in brewing process and final product quality.

Gugino, I. M., Alfeo, V., Ashkezary, M. R., Marconi, O., Pirrone, A., Francesca, N., Cincotta, F., Verzera, A. & Todaro, A. (2024). Food Chemistry, 435, 137517.

This study explores the potential of Maiorca wheat malt as an alternative ingredient in beer production, investigating its impact on the brewing process and beer quality at different recipe contents (50 %, 75 %, 100 %). The study encompasses a comprehensive analysis of key malt parameters, revealing Maiorca malt’s positive influence on maltose, glucose, filterability, extract, free amino nitrogen, and fermentability. Notably, the malt exhibited heightened levels of α-amylase and β-amylase enzymes compared to conventional commercial malt. Furthermore, the analysis of aroma compounds and subsequent sensory evaluations unveiled a significant correlation between the proportion of Maiorca malt in the formulation and intensified estery, fruity, malty, honey, complemented by a reduction in attributes such as aromatic compounds, phenolic, yeasty, sulfury, oxidized, and solvent-like odors. This research underscores the favorable contribution of Maiorca wheat malt to enhancing both the brewing process and final beer quality, highlighting its potential as an innovative ingredient in brewing practices.

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Safety Information
Symbol : GHS05, GHS08
Signal Word : Danger
Hazard Statements : H314, H315, H319, H334
Precautionary Statements : P260, P261, P264, P280, P284, P301+P330+P331, P302+P352, P303+P361+P353, P304+P340
Safety Data Sheet
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