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D-Glucose Assay Kit (GOPOD Format)

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00:03  Introduction
00:46   Principle
01:40   Reagent Preparation
03:41    Sample Preparation
05:16    Procedure
07:17    Calculations

D-Glucose Assay Kit GOPOD Format K-GLUC Scheme
   
Product code: K-GLUC
€217.00

660 assays per kit

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Available for shipping

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Content: 660 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: D-Glucose
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 510
Signal Response: Increase
Linear Range: 4 to 100 μg of glucose per assay
Limit of Detection: 40 mg/L
Reaction Time (min): ~ 20 min
Application examples: Wine, beer, fruit juices, soft drinks, milk, jam, dietetic foods, bakery products, candies, fruit and vegetables, tobacco, cosmetics, pharmaceuticals, feed, paper and other materials (e.g. biological cultures, samples, etc.).
Method recognition: Widely used and accepted in clinical chemistry and food analysis

The D-Glucose test kit contains high purity reagents for the measurement and analysis of D-glucose in cereal extracts and for use in combination with other Megazyme kits.

See more related mono/disaccharide assay kit products.

Scheme-K-GLUC GLUC Megazyme

Advantages
  • All reagents stable for > 12 months after preparation 
  • Very competitive price (cost per test) 
  • Simple format 
  • Standard included
Documents
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance Validation Report
Publications
Megazyme publication

Diastatic power and maltose value: a method for the measurement of amylolytic enzymes in malt.

Charmier, L. M., McLoughlin, C. & McCleary, B. V. (2021). Journal of the Institute of Brewing, In Press.

A simple method for measurement of the amylolytic activity of malt has been developed and fully evaluated. The method, termed the Maltose Value (MV) is an extension of previously reported work. Here, the MV method has been studied in detail and all aspects of the assay (sample grinding and extraction, starch hydrolysis, maltose hydrolysis and determination as glucose) have been optimised. The method is highly correlated with other dextrinising power methods. The MV method involves extraction of malt in 0.5% sodium chloride at 30°C for 20 minutes followed by filtration; incubation of an aliquot of the undiluted filtrate with starch solution (pH 4.6) at 30°C for 15 min; termination of reaction with sodium hydroxide solution; dilution of sample in an appropriate buffer; hydrolysis of maltose with a specific α-glucosidase; glucose determination and activity calculation. Unlike all subsequent reducing sugar methods, the maltose value method measures a defined reaction product, maltose, with no requirement to use equations to relate analytical values back to Lintner units. The maltose value method is the first viable method in 130 years that could effectively replace the 1886 Lintner method.

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Megazyme publication

Measurement of available carbohydrates in cereal and cereal products, dairy products, vegetables, fruit and related food products and animal feeds: First Action 2020.07.

McCleary, B. V. & McLoughlin, C. (2021). Journal of AOAC International, qsab019.

Background: The level of available carbohydrates in our diet is directly linked to two major diseases; obesity and Type II diabetes. Despite this, to date there is no method available to allow direct and accurate measurement of available carbohydrates in human and animal foods. Objective: The aim of this research was to develop a method that would allow simple and accurate measurement of available carbohydrates, defined as non-resistant starch, maltodextrins, maltose, isomaltose, sucrose, lactose, glucose, fructose and galactose. Method: Non-resistant (digestible) starch is hydrolysed to glucose and maltose by pancreatic α-amylase and amyloglucosidase at pH 6.0 with shaking or stirring at 37°C for 4 h. Sucrose, lactose, maltose and isomaltose are completely hydrolyzed by specific enzymes to their constituent monosaccharides, which are then measured using pure enzymes in a single reaction cuvette. Results: A method has been developed that allows the accurate measurement of available carbohydrates in all cereal, vegetable, fruit, food, and feed products, including dairy products. Conclusions: A single-laboratory validation was performed on a wide range of food and feed products. The inter-day repeatability (%RSDr) was <3.58% (w/w) across a range of samples containing 44.1 to 88.9% available carbohydrates. The LOD and LOQ obtained were 0.054% (w/w) and 0.179% (w/w), respectively. The method is all inclusive, specific, robust and simple to use. Highlights: A unique method has been developed for the direct measurement of available carbohydrates, entailing separate measurement of glucose, fructose and galactose; information of value in determining the glycemic index of foods.

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Megazyme publication

Measurement of Starch: Critical evaluation of current methodology.

McCleary, B. V., Charmier, L. M. J. & McKie, V. A. (2018). Starch‐Stärke, 71(1-2), 1800146.

Most commonly used methods for the measurement of starch in food, feeds and ingredients employ the combined action of α‐amylase and amyloglucosidase to hydrolyse the starch to glucose, followed by glucose determination with a glucose oxidase/peroxidase reagent. Recently, a number of questions have been raised concerning possible complications in starch analytical methods. In this paper, each of these concerns, including starch hydrolysis, isomerisation of maltose to maltulose, effective hydrolysis of maltodextrins by amyloglucosidase, enzyme purity and hydrolysis of sucrose and β‐glucans have been studied in detailed. Results obtained for a range of starch containing samples using AOAC Methods 996.11 and 2014 .10 are compared and a new simpler format for starch measurement is introduced. With this method that employs a thermostable α-amylase (as distinct from a heat stable α-amylase) which is both stable and active at 100°C and pH 5.0, 10 samples can be analysed within 2 h, as compared to the 6 h required with AOAC Method 2014.10.

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Megazyme publication

Megazyme “advanced” wine test kits general characteristics and validation.

Charnock, S. J., McCleary, B. V., Daverede, C. & Gallant, P. (2006). Reveue des Oenologues, 120, 1-5.

Many of the enzymatic test kits are official methods of prestigious organisations such as the Association of Official Analytical Chemicals (AOAC) and the American Association of Cereal Chemists (AACC) in response to the interest from oenologists. Megazyme decided to use its long history of enzymatic bio-analysis to make a significant contribution to the wine industry, by the development of a range of advanced enzymatic test kits. This task has now been successfully completed through the strategic and comprehensive process of identifying limitations of existing enzymatic bio-analysis test kits where they occurred, and then using advanced techniques, such as molecular biology (photo 1), to rapidly overcome them. Novel test kits have also been developed for analytes of emerging interest to the oenologist, such as yeast available nitrogen (YAN; see pages 2-3 of issue 117 article), or where previously enzymes were simply either not available, or were too expensive to employ, such as for D-mannitol analysis.

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Megazyme publication

Grape and wine analysis: Oenologists to exploit advanced test kits.

Charnock, S. C. & McCleary, B. V. (2005). Revue des Enology, 117, 1-5.

It is without doubt that testing plays a pivotal role throughout the whole of the vinification process. To produce the best possible quality wine and to minimise process problems such as “stuck” fermentation or troublesome infections, it is now recognised that if possible testing should begin prior to harvesting of the grapes and continue through to bottling. Traditional methods of wine analysis are often expensive, time consuming, require either elaborate equipment or specialist expertise and frequently lack accuracy. However, enzymatic bio-analysis enables the accurate measurement of the vast majority of analytes of interest to the wine maker, using just one piece of apparatus, the spectrophotometer (see previous issue No. 116 for a detailed technical review). Grape juice and wine are amenable to enzymatic testing as being liquids they are homogenous, easy to manipulate, and can generally be analysed without any sample preparation.

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Publication

Mechanistic insights into the enhanced texture of potato noodles by incorporation of small granule starches.

Ma, M., Zhang, X., Zhu, Y., Li, Z., Sui, Z. & Corke, H. (2024). International Journal of Biological Macromolecules, 257, 128535.

Potato noodles are a popular food due to their unique texture and taste, but native potato starch often fails to meet consumer demands for precise textural outcomes. The effect of blending small granule (waxy amaranth, non-waxy oat and quinoa) starch with potato starch on the properties of noodles was investigated to enhance quality of noodles. Morphological results demonstrated that small granule starch filled gaps between potato starch granules, some of which gelatinized incompletely. Meanwhile, XRD and FTIR analysis indicated that more ordered structures and hydrogen bonding among starch granules increased with addition of small granule starch. The addition of oat or quinoa starch increased gel elasticity, decreased viscosity of the pastes, and increased the tensile strength of noodles, while addition of 30 % and 45 % waxy amaranth starch did not increase G′ value of gel or tensile strength of noodles. These results indicated that amylose molecules played an important role during retrogradation, and may intertwine and interact with each other to enhance the network structure of starch gel in potato starch blended with oat or quinoa starch. This study provides a natural way to modify potato starch for desirable textural properties of noodle product.

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Publication

Effects of single and dual modifications with debranching and heat-moisture treatments on physicochemical, rheological, and digestibility properties of proso millet starch.

Kumar, S. R., Tangsrianugul, N., Sriprablom, J., Winuprasith, T., Wansuksri, R. & Suphantharika, M. (2023). Carbohydrate Polymer Technologies and Applications, 6, 100399.

The effects of single and dual modifications with heat-moisture treatment (HMT) and enzymatic debranching on the structural, physicochemical, rheological, and in-vitro digestibility properties of proso millet starch (PMS) were comparatively studied. Amylose content increased after gelatinization and all modification treatments. Debranching and dual modification (debranching followed by HMT) decreased the average chain length and swelling power but increased the solubility of PMS. HMT did not change the average chain length and chain length distribution in native and debranched starches but decreased the swelling power and solubility of native starch. The A-type X-ray diffraction pattern of PMS was converted to the A + V type by HMT and to the B + V type by debranching and dual modification. FTIR investigation showed that the short-range molecular order of all modified starches was disrupted. The viscoelasticity and viscosity of all starch pastes decreased in the following order: PMS > HMT starch > debranched starch > dual modified starch. In-vitro starch digestibility showed that HMT increased slowly digestible starch (SDS) to 32.88 %, while debranching increased resistant starch (RS) to 77.14 %. HMT increased RS to 86.17 % in the dual modified starch. Debranching therefore predominates in the dual modification of starch compared to HMT.

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Publication

Effects of heat–moisture treatment on structural characteristics and in vitro digestibility of A-and B-type wheat starch.

Wang, J., Huang, J., Liang, Q. & Gao, Q. (2024). International Journal of Biological Macromolecules256, 128012.

In this study, A- and B-type wheat starch granules (AWS and BWS) were separated and modified by heat-moisture treatment (HMT) with different moisture content (10 %–40 %). The effects of HMT on the structure characteristics and digestibility of raw/cooked AWS and BWS were investigated by SEM, FT-IR, XRD, DSC, TGA and NMR. SEM and FT-IR results showed that BWS was more sensitive to HMT than AWS. Interestingly, crystalline conformation of AWS and BWS changed from A type to A + V type after HMT, and the relative crystallinity (V-type) of starch increased to 2.7 % and 3.4 %, respectively. XRD and NMR results verified the formation of V-type crystalline structure. The resistant starch (RS) content of cooked starch was increased, especially for BWS (from 11.46 % to 28.29 %). Compared to the cooked starch, the RS content of raw AWS and BWS was affected by relative crystallinity and the size of starch granules. Furthermore, structure characteristics and digestion kinetics results indicated that the digestion rate of cooked AWS increased due to the deconstruction of starch chains, opposite to BWS (because of the more V-type crystals). The results enrich our understanding of the mechanism of digestion subjected to HMT by different grain sizes of the same wheat starch.

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Publication

Characterisation of physicochemical parameters and antibacterial properties of New Caledonian honeys.

Bucekova, M., Godocikova, J., Gueyte, R., Chambrey, C. & Majtan, J. (2023). Plos one, 18(10), e0293730.

Honey is an attractive natural product with various health benefits. A few honey-based commercial products have successfully been adopted in clinics to improve wound healing. However, screening of other potential sources of medical-grade honey, in particular, honeys from territories with high floral species diversity and high endemicity, is highly needed. The goal of this study was to characterise the physicochemical and antibacterial properties of New Caledonian honey samples (n = 33) and to elucidate the major mechanism of their antibacterial action. Inhibitory antibacterial activity of honeys against Staphylococcus aureus and Pseudomonas aeruginosa was determined with a minimum inhibitory concentration (MIC) assay. Enzymatic activity of glucose oxidase and the content of hydrogen peroxide (H2O2) in honey samples were analysed. Furthermore, total protein content of honeys together with their electrophoretic protein profiles were also determined in the study. The antibacterial efficacy of 24% of the tested honey samples was slightly superior to that of manuka honey with unique manuka factor 15+. The antibacterial activity of catalase-treated honey sample solutions was significantly reduced, suggesting that H2O2 is a key antibacterial compound of diluted honeys. However, the kinetic profiles of H2O2 production in most potent honeys at a MIC value of 6% was not uniform. Under the experimental conditions, we found that a H2O2 concentration of 150 μM in diluted honeys is a critical concentration for inhibiting the growth of S. aureus. In contrast, 150 μM H2O2 in artificial honey solution was not able to inhibit bacterial growth, suggesting a role of phytochemicals in the antibacterial activity of natural honey. In addition, the continuous generation of H2O2 in diluted honey demonstrated an ability to counteract additional bacteria in re-inoculation experiments. In conclusion, the tested New Caledonian honey samples showed strong antibacterial activity, primarily based on H2O2 action, and therefore represent a suitable source for medical-grade honey.

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Publication

Lipid complexation reduces rice starch digestibility and boosts short-chain fatty acid production via gut microbiota.

Shen, Y., An, Z., Huyan, Z., Shu, X., Wu, D., Zhang, N., Pellegrini, N. & Rubert, J. (2023). npj Science of Food, 7(1), 56.

In this study, two rice varieties (RS4 and GZ93) with different amylose and lipid contents were studied, and their starch was used to prepare starch-palmitic acid complexes. The RS4 samples showed a significantly higher lipid content in their flour, starch, and complex samples compared to GZ93. The static in vitro digestion highlighted that RS4 samples had significantly lower digestibility than the GZ93 samples. The C of the starch-lipid complex samples was found to be 17.7% and 18.5% lower than that of the starch samples in GZ93 and RS4, respectively. The INFOGEST undigested fractions were subsequently used for in vitro colonic fermentation. Short-chain fatty acids (SCFAs) concentrations, mainly acetate, and propionate were significantly higher in starch-lipid complexes compared to native flour or starch samples. Starch-lipid complexes produced a distinctive microbial composition, which resulted in different gene functions, mainly related to pyruvate, fructose, and mannose metabolism. Using Model-based Integration of Metabolite Observations and Species Abundances 2 (MIMOSA2), SCFA production was predicted and associated with the gut microbiota. These results indicated that incorporating lipids into rice starch promotes SCFA production by modulating the gut microbiota selectively.

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Publication

Intensification of corn fiber saccharification using a tailor made enzymatic cocktail.

Manso, J. O., Nielsen, M. B., Moya, E. B., Sandri, J. P., Yamakawa, C. K. & Mussatto, S. I. (2024). Enzyme and Microbial Technology172, 110347.

The transition from an economic model based on resource extraction to a more sustainable and circular economy requires the development of innovative methods to unlock the potential of raw materials such as lignocellulosic biomasses. Corn fiber differs from more traditional lignocellulosic biomasses due to its high starch content, which provides additional carbohydrates for fermentation-based biomanufacturing processes. Due to its unique chemical composition, this study focused on the development of a tailor made enzymatic cocktail for corn fiber saccharification into monosaccharides. Three commercially available hydrolytic enzymes (Cellic® CTec2, Pentopan® Mono BG, and Termamyl® 300 L) were combined to hydrolyze the polysaccharide structure of the three main carbohydrate fractions of corn fiber (cellulose, hemicellulose and starch, respectively). Prior to saccharification, corn fiber was submitted to a mild hydrothermal pretreatment (30 min at 100 °C). Then, two experimental designs were used to render an enzymatic cocktail capable of providing efficient release of monosaccharides. Using 60 FPU/g DM of Cellic® CTec2 and 4.62 U/g DM of Termamyl® 300 L, without addition of Pentopan® Mono BG, resulted in the highest efficiencies for glucose and xylose release (66% and 30%, respectively). While higher enzyme dosages could enhance the saccharification efficiency, adding more enzymes would have a more pronounced effect on the overall process costs rather than in increasing the efficiency for monosaccharides release. The results revealed that the recalcitrance of corn fiber poses a problem for its full enzymatic degradation. This fact combined with the unique chemical composition of this material, justify the need for developing a tailor made enzymatic cocktail for its degradation. However, attention should also be given to the pretreatment step to reduce even more the recalcitrance of corn fiber and improve the performance of the tailored cocktail, as a consequence.

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Publication

The role of herbal teas in reducing the starch digestibility of cooked rice (Oryza sativa L.): An in vitro co-digestion study.

Aumasa, T., Ogawa, Y., Singh, J., Panpipat, W. & Donlao, N. (2023). NFS Journal, 33, 100154.

Herbal teas are well known for their antidiabetic effects due to the abundance of phenolic acids, flavonoids, and tannins. An in vitro co-digestion test was conducted to observe influence of herbal teas (HTs) i.e., beal fruit (BA), mulberry leaf (MB), gymnema leaf (GM), and chrysanthemum flower (CS) on starch digestibility of cooked rice and to elucidate correlation analysis of phytochemicals and their antioxidant activities during in vitro starch hydrolysis (SH). HTs prepared from GM and MB showed highest reduction of SH, kinetic constant and estimated glycemic index (eGI) of cooked rice, followed by BA and CS. Besides, MB and GM teas decreased the eGI of cooked rice up to 15%, followed by BA and CS teas, respectively. Phytochemicals and their antioxidant activities were positively correlated to SH, with both parameters displaying a higher decrease in intestinal phase when compared to gastric phase during in vitro co-digestion. These results indicate a strong interaction among phytochemicals and α-amylase in influencing glycaemic parameters. Thus, HTs has the potential to provide functional health benefits in lowering postprandrial hyperglycemia.

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Publication

Growth-coupled anaerobic production of isobutanol from glucose in minimal medium with Escherichia coli.

Boecker, S., Schulze, P. & Klamt, S. (2023). Biotechnology for Biofuels and Bioproducts, 16(1), 148.

Background: The microbial production of isobutanol holds promise to become a sustainable alternative to fossil-based synthesis routes for this important chemical. Escherichia coli has been considered as one production host, however, due to redox imbalance, growth-coupled anaerobic production of isobutanol from glucose in E. coli is only possible if complex media additives or small amounts of oxygen are provided. These strategies have a negative impact on product yield, productivity, reproducibility, and production costs. Results: In this study, we propose a strategy based on acetate as co-substrate for resolving the redox imbalance. We constructed the E. coli background strain SB001 (ΔldhA ΔfrdA ΔpflB) with blocked pathways from glucose to alternative fermentation products but with an enabled pathway for acetate uptake and subsequent conversion to ethanol via acetyl-CoA. This strain, if equipped with the isobutanol production plasmid pIBA4, showed robust exponential growth (µ = 0.05 h−1) under anaerobic conditions in minimal glucose medium supplemented with small amounts of acetate. In small-scale batch cultivations, the strain reached a glucose uptake rate of 4.8 mmol gDW−1 h−1, a titer of 74 mM and 89% of the theoretical maximal isobutanol/glucose yield, while secreting only small amounts of ethanol synthesized from acetate. Furthermore, we show that the strain keeps a high metabolic activity also in a pulsed fed-batch bioreactor cultivation, even if cell growth is impaired by the accumulation of isobutanol in the medium. Conclusions: This study showcases the beneficial utilization of acetate as a co-substrate and redox sink to facilitate growth-coupled production of isobutanol under anaerobic conditions. This approach holds potential for other applications with different production hosts and/or substrate–product combinations.

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Publication

Starch molecular structures in relation to properties of ratoon rice produced by different ratooning practices.

Yang, X., Peng, T., Xu, Y., Gao, K., Zhao, Q. & Song, X. (2024). Carbohydrate Polymers, 323, 121459.

The development of forage-grain ratoon rice (RR) pattern could ensure food security and promote silage production. Herein three indica rice varieties were used to investigate the influence of different forage clipping stages (heading, milk-ripe, wax-ripe, and full-ripe) on starch molecular structures and RR properties. The apparent amylose contents (AAC) of starches increased, but pasting viscosities, gelatinization temperatures and starch sizes decreased with the postponement of clipping stages due to the retardation of endosperm development. The starches showed A-type crystalline structure with increased in vitro digestibility; however relative crystallinity decreased by 13.45 % to 23.89 %. The short fa (DP 6-12) chains of amylopectin increased while long fb3 (DP ≥ 37) chains decreased (p < 0.05). The proportions of amylose chains with DP 100-2000 increased but those with DP 2000-20,000 decreased. Rice grain strength was positively correlated with fb3 chains while negatively correlated with fa chain. The hardness of cooked RR was positively correlated with AAC while negatively correlated with fb2 (DP 25–36). RR clipping at milk-ripe stage had the highest grain strength and moderate texture properties. The elucidation of structure-property relationships is helpful for RR utilization and development of suitable cultivation conditions for RR production.

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Publication

Recovery of Nanocellulose from Agri-Food Residues through Chemical and Physical Processes.

Pirozzi, A., Pappalardo, G. & Donsì, F. (2023). Chemical Engineering Transactions, 102, 175-180.

This work proposes a biorefinery approach for the exploitation of agri-food by-products, such as tomato pomace (TP), through the combination of mild chemical hydrolysis and high-pressure homogenization (HPH) in water not only to promote the recovery of cellulose but also its defibrillation to obtain nanocellulose. In particular, the cellulose pulp was isolated from TP using different combinations of chemical and physical processes, by applying HPH treatment (i) directly on the raw material, (ii) after the acid hydrolysis, and (iii) after alkaline hydrolysis. Moreover, the isolated cellulose was deconstructed to obtain cellulose nanoparticles, also through the application of the HPH treatment, enhancing the polymer properties. The structural and physical features of cellulose nanoparticles from TP were analyzed through Fourier-transform infrared spectroscopy (FT-IR) analysis, ?-potential measurement, and morphological analysis with SEM. The results clearly showed that the HPH treatment (80 MPa, 20 passes) at different stages of the process caused only a slight increase in the yield of cellulose recovery, but significantly contributed to obtaining defibrillated cellulose particles, characterized by smaller irregular domains containing elongated needle-like fibers.

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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, P303+P361+P353
Safety Data Sheet
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