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Fructan HK Assay Kit

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00:02  Theory of the Analytical Procedure
02:07   Kit Description
03:12    Preparation of Reagent Solutions/Suspensions
05:30   Samples Containing 0–12% Fructan
07:55    Samples Containing 12–100% Fructan
10:07     Analysis of Fructan Content: a. Hydrolysis of Sucrose & Low DP Maltosaccharides
12:39     Analysis of Fructan Content: b. Hydrolysis of Fructan
14:16      Analysis of Fructan Content: c. Measurement of Fructan
17:14      Calculation of Fructan Content

Fructan HK Assay Kit K-FRUCHK Scheme
Product code: K-FRUCHK

50 assays per kit

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Content: 50 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: Fructan
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Increase
Linear Range: 4 to 80 µg of D-glucose, D-fructose or sucrose per assay
Limit of Detection: 1 g/100 g
Total Assay Time: ~ 30 min
Application examples: Flours, plant materials (e.g. onion), food products and other materials

The Fructan HK test kit is suitable for the specific measurement and analysis of all fructo-oligosaccharides (reducing and non-reducing) and of fructan polysaccharides but is not suitable for the analysis of samples containing high levels of D-glucose, D-fructose, sucrose or maltose.

View our full range of assay kits for polysaccharides.

Scheme-K-FRUCHK FRUCHK Megazyme

  • Very cost effective 
  • All reagents stable for > 12 months after preparation 
  • Fructan kits are available only from Megazyme 
  • Simple format 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Standard included
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance
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

Measurement of total fructan in foods by enzymatic/spectrophotometric method: Collaborative study.

McCleary, B. V., Murphy, A. & Mugford, D. C. (2000). Journal of AOAC International, 83(2), 356-364.

An AOAC collaborative study was conducted to evaluate the accuracy and reliability of an enzyme assay kit procedure for measuring oligofructans and fructan polysaccharide (inulins) in mixed materials and food products. The sample is extracted with hot water, and an aliquot is treated with a mixture of sucrase (a specific sucrose-degrading enzyme), α-amylase, pullulanase, and maltase to hydrolyze sucrose to glucose and fructose, and starch to glucose. These reducing sugars are then reduced to sugar alcohols by treatment with alkaline borohydride solution. The solution is neutralized, and excess borohydride is removed with dilute acetic acid. The fructan is hydrolyzed to fructose and glucose using a mixture of purified exo- and endo-inulinanases (fructanase mixture). The reducing sugars produced (fructose and glucose) are measured with a spectrophotometer after reaction with para-hydroxybenzoic acid hydrazide. The samples analyzed included pure fructan, chocolate, low-fat spread, milk powder, vitamin tablets, onion powder, Jerusalem artichoke flour, wheat stalks, and a sucrose/cellulose control flour. Repeatability relative standard deviations ranged from 2.3 to 7.3%; reproducibility relative standard deviations ranged from 5.0 to 10.8%.

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Megazyme publication

Measurement of inulin and oligofructan.

McCleary, B. V. & Blakeney, A. B. (1999). Cereal Foods World, 44, 398-406.

Fructans are defined as any compound in which one or more fructosyl-fructose linkages constitute a majority of the linkages (1). This refers to polymeric material as well as to oligomers as small as disaccharide inulobiose. Fructans are widely distributed in the plant kingdom. They are present in monocotyledons, dicotyledons, and green algae. Fructans differ in molecular structure and in molecular weight. They may be classified into three main types, the inulin type, the levan (previously called phlein) type, and the graminan type (2). The inulin group consists of material that has mostly of exclusively the (2-1) fructosly-fructose linkage. Levan is material that contains mostly or exclusively the (2-6) fructosyl-fructose linkage. The graminan (or branched) type has both (2-1) and (2-6) fructosly-fructose linkages in significant amounts (e.g. graminan from Gramineae). The distribution of fructans in nature, and the production of fructooligosaccharides, such as neosugar, using fructosyltransferase, has been reviewed in a monograph (3). In the context of this article and the analytical procedure described, the term fructan relates only to inulin and graminan. The current analytical procedure has not been evaluated on levan.

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Megazyme publication

Measurement of inulin and inulin-degrading enzymes.

McCleary, B. V. (1998). “Proceedings of the Seventh Seminar on Inulin”, (A. Fuchs and A. Van Laere, Eds.), European Fructan Association, pp. 36-45.

A non-instrumental method for the measurement of fructan is described. The method simplifies fructan analysis, is easy to perform, uses standard laboratory equipment, and is accurate, reproducible and specific. The procedure employs highly purified and specific enzymes to hydrolyse sucrose, starch and fructans (inulins and graminan).

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Megazyme publication

Fructans - Analytical approaches to a fibre that ferments.

Blakeney, A. B., McCleary, B. V. & Mugford, D. C. (1997). Chemistry in Australia, 17-19.

Fructans are defined as any compound where one or more fructosyl-fructose linkages constitute a majority of the linkages. This refers to polymeric material as well as oligomers as small as the diasaccharide inulobiose. Material included in this definition may or may not contain attached glucose. The terms oligomer and polymer are used by fructan researchers to distinguish between materials that can be specifically characterised and those that can not. Fructans are widely distributed in the plant kingdom. They are present in monocotyledons, dicotyledons and in green algae.

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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
Enzymic analysis of the fine structure of galactomannans.

McCleary, B. V. (1994). “Methods in Carbohydrate Chemistry”, Vol. X, (J. N. BeMiller, D. J. Manners and R. J. Sturgeon, Eds.), John Wiley & Sons Inc., pp. 175-182.

A number of methods have been described for the analysis of the fine structure of galactomannans, i.e., the distribution of D-galactosyl units along the D-mannan backbone (1). Such studies include the analysis of x-ray diffraction data of stretched fibers of galactomannans (2,3), 1H- and 13C-nmr (nuclear magnetic resonance) of native and partially depolymerized galacto¬mannans (4) and a range of chemical procedures (5-7), including those employing a detailed theoretical analysis of the kinetics of reaction (8). An alternative approach involves the characterization and quantification of the oligosaccharides produced on hydrolysis of galactomannans by highly purified and well-characterized β-mannanases (EC (9,10). The β-mannanases employed were purified to homogeneity by affinity chromatography on gIucornannan-AH-Sepharose 4B. They were characterized by a range of physicochemicai procedures by determining the kinetics of their action on β-mannooligosaccharides, and by characterizing the structures of oligosaccharides produced on hydrolysis of galactomannans and glucomannans (11). From these studies, a basic model describing the subsite binding requirements of all the β-mannanases examined was proposed (Fig. 1). This model was then modified to account for the slight differences noted in the types of oligosaccharides produced by β-mannanases from different sources. The β-mannanases which differ most significantly in their action patterns on galactomannans are those from Aspergillus niger culture filtrates and from germinated guar seed.

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Rheological properties and characterisation of some bioactive components in flours made of different coloured sweet potato (Ipomoea batatas L.) genotypes.

Mihály-Langó, B., Ács, K., Berényi, A., Maróti Tóth, K., Táborosi Ábrahám, Z., Gáll, T. & Ács, E. (2023). Acta Alimentaria, 52(4), 570-578.

The popularity of sweet potatoes in Central Europe has been increasing recently, mainly the high-quality, perfect, fresh tubers are in demand. However, out of class grade tubers could be marketed in dried, grounded form as sweet potato flour. The aim of this study was to characterise some important nutritional properties of flours of three sweet potato genotypes with different tuber colours (white, purple, and orange) and to investigate how this raw material affects the rheological properties of sweet potato-wheat flour blends. Dietary fibres are present in sweet potatoes in a significant proportion, orange coloured flour showed the highest values. The main free sugars were sucrose, glucose, and fructose, but sucrose was the dominant one. Antioxidant capacity and total phenolic content also varied considerably, the purple flour had the highest values. Mineral composition showed significant variability, the purple flour contained the highest level of minerals. It was confirmed that adding sweet potato flour to wheat flour affected its rheological properties, however in a varied manner. For the orange flour these properties have lightly decreased, though it had no significant effect on dough quality, while the white and purple flours with a dosage of 5, 10 and 15% could improve the dough behaviour. Thus, sweet potato in this form is a valuable raw material.

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Alteration of Carbohydrate Metabolism in Fusarium Infected Wheat Kernels Treated with Fungicides and Its Relation to Baking Technological Parameters and Deoxynivalenol Contamination.

Acs, K., Varga, M., Szekeres, A., Salgo, A., Lantos, C., Bekes, F., Pauk, J. & Mesterhazy, A. (2023). Agriculture, 13(4), 868.

Changes of water-soluble carbohydrate (WSC) content such as fructose, glucose, sucrose, maltose, nystose, raffinose, stachyose and fructan were analyzed in wheat kernels in Fusarium epidemic and non-epidemic seasons. In both season types, eight commercial fungicides were applied and three wheat varieties with differing Fusarium resistance were tested. In the epidemic year, the average total amount of WSC was above 1.6% which was 2 times higher than in the non-epidemic year (0.7%). Sucrose, maltose, raffinose and fructan components determined the increased WSC value, but the most substantial change was observed in maltose content where its average amount was 28 times higher in the epidemic year. Fungicide application also significantly increased all the carbohydrate components except maltose, where significant reduction was observed. WSC components had strong correlation with several farinograph or extensograph parameters, but only the maltose content showed positive strong correlation (r = 0.9) with deoxynivalenol (DON) toxin that was highly affected by the applied fungicide. The changes of WSC indicate altered carbohydrate synthesis along with abnormal degradation processes and thus have impaction on the baking features. It seems that the sugar metabolism interacts with DON synthesis and the results give important additional information to the altered metabolism of the attacked plant.

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Fibre and short-chain carbohydrate composition in rye varieties, novel industrial milling fractions and breads.

Szentmiklóssy, M. K. J., Jaksics, E., Farkas, A., Pusztai, É., Kemény, S., Németh, R. & Tömösközi, S. (2023). Acta Alimentaria, 52(2), 177-189.

Rye is an important raw material of bread due to tradition and its favourable nutritional and technological qualities. Despite the beneficial fibre composition, a special group of short-chain carbohydrates, the so called FODMAPs (fermentable oligo-, di-, monosaccharides and polyols) may cause problems for patients with irritable bowel syndrome. The aim of our work was to investigate the non-starch carbohydrate (dietary fibre compounds, short-chain carbohydrates) composition of rye varieties, and of their novel milling fractions obtained from industrial milling trials and test loaves made from them. Regarding fibre and short chain carbohydrate composition, rye varieties did not show significant differences. In new subfractions, fibre and FODMAP composition were described, among profiles most of them differ from commonly used flours, independently from variety. The yeast fermentation and baking caused a decrease in water-extractable arabinoxylan content, at the same time increased the substitution pattern of water-extractable arabinoxylans. Furthermore, breadmaking process decreased the fructan content, and therefore increased the fructose level, thus modifying the short-chain carbohydrate composition. Based on our knowledge, this research is among the first ones investigating the fibre and short-chain composition of rye from the seeds to the consumable final products.

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FODMAP modulation as a dietary therapy for IBS: Scientific and market perspective.

Ispiryan, L., Zannini, E. & Arendt, E. K. (2022). Comprehensive Reviews in Food Science and Food Safety, 21(2), 1491-1516.

A diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAPs) is a promising therapeutic approach to reduce gastrointestinal symptoms associated with irritable bowel syndrome (IBS). However, a shift toward a more sustainable, healthy diet with higher inclusion of whole-grain cereals (i.e., wheat, rye, barley) and pulses, naturally rich in FODMAPs, poses a severe challenge for susceptible individuals. Dietary restriction of fermentable carbohydrates (commonly called the "low FODMAP diet") has received significant consideration. Hence, the development of functional low FODMAP products is emerging in food science and the food industry. In this review, we evaluate the most promising yet neglected (bio)-technological strategies adopted for modulating the FODMAP contents in complex food systems and the extent of their uptake in the global food market. We extensively investigated the global low FODMAP market, contrasted with the status quo in food science and discussed the key principles and concomitant challenges of targeted FODMAP reduction strategies. Powerful tools are available which are based either on the use of ingredients where FODMAPs have been physically removed (e.g., by membrane filtration) or biotechnologically reduced during the food processing, mediated by added enzymes, microbial enzymes during a fermentation process, and seed endogenous enzymes. However, <10% of the small market of functional products with a low FODMAP claim (total ~800 products) used any of the targeted FODMAP reduction techniques. The global market is currently dominated by gluten-free products, which are naturally low in FODMAPs and characterized by inferior sensory attributes.

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Tritordeum breads are well tolerated with preference over gluten‐free breads in non‐celiac wheat‐sensitive patients and its consumption induce changes in gut bacteria.

Sánchez‐León, S., Haro, C., Villatoro, M., Vaquero, L., Comino, I., González‐Amigo, A. B., Vivas, S., Pastor, J., Sousa, C., Landa, B. B. & Barro, F. (2021). Journal of the Science of Food and Agriculture, 101(8), 3508-3517.

Background: The ingestion of wheat and other cereals are related to several gut disorders. The specific components responsible for non-celiac wheat-sensitivity (NCWS) may include gluten and other compounds. Tritordeum is a new cereal derived from crossing durum wheat with a wild barley species, which differs from bread wheat in its gluten composition. In the present work, we examined the response of NCWS patients to tritordeum bread Gastrointestinal symptoms as well as tritordeum acceptability, gluten immunogenic peptides excretion, and the composition and structure of the intestinal microbiota were evaluated. Results: Gastrointestinal symptoms of the subjects showed no significant change between the gluten-free bread and the tritordeum bread. Participating subjects rated tritordeum bread higher than the gluten-free bread. Analysis of the bacterial gut microbiota indicated that tritordeum consumption does not alter the global structure and composition of the intestinal microbiota, and only a few changes in some butyrate-producing bacteria were observed. Conclusions: All the results derived from acceptability, biochemical and microbiological tests suggest that tritordeum may be tolerated by a sub-set of NCWS sufferers who do not require strict exclusion of gluten from their diet.

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Reducing FODMAPs and improving bread quality using type II sourdough with selected starter cultures.

Menezes, L. A. A., De Marco, I., Neves Oliveira dos Santos, N., Costa Nunes, C., Leite Cartabiano, C. E., Molognoni, L., de Melo Pereira, G. V., Daguer, H. & De Dea Lindner, J. (2021). International Journal of Food Sciences and Nutrition, 1-11.

This study focussed on lactic acid bacteria (LAB) screening for sourdough type II elaboration and evaluating the effects of sourdough fermentation in bread making, focussing mainly on reducing FODMAPs. After a technological performance screening, six strains (Levilactobacillus brevisWeissella minorLactiplantibacillus plantarumLeuconostoc citreumLimosilactobacillus fermentum, and Companilactobacillus farciminis) were selected for sourdough preparation. Total titratable acidity, pH, specific volume, and enumeration of microorganisms were carried out on sourdoughs, doughs, and breads. Breads were subjected to texture profile and colour analysis, moulds and yeast enumeration, and total fructans (main group of FODMAPs) quantification. Breads produced with sourdough showed a significant reduction of fructans, greater acidity, volume, and better performance during storage when compared to fermentation using only baker's yeast. Including specific cultures as starters in sourdough reduced fructans content by >92%, thereby producing a low FODMAP bread suitable for Irritable Bowel Syndrome patients with improved nutritional and technological properties.

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β-fructosidase FosE activity in Lactobacillus paracasei regulates fructan degradation during sourdough fermentation and total FODMAP levels in steamed bread.

Fang, S., Yan, B., Tian, F., Lian, H., Zhao, J., Zhang, H., Chen, W. & Fan, D. (2021). LWT, 145, 111294.

Low fermentable oligo-, di- and monosaccharides, and polyols (FODMAPs) have been previously shown to effectively relieve symptoms of irritable bowel syndrome (IBS) patients. This study aimed to evaluate the effect of lactobacilli on the FODMAPs degradation during sourdough fermentation, especially on fructan. Phylogenomic analysis of lactobacilli revealed that the gene coding for β-fructosidase FosE are present in Lactobacillus paracasei. L. paracasei FJSSZ3L1 was selected to effectively degrade fructans, with a degradation percentage of 86.57 ± 2.88%. The β-fructosidase FosE activity with substrate specificity was detected in the supernatant and cell wall extract, and the location of enzyme activity was related to the growth time of the strain. Fructan content decreased during sourdough fermentation with L. paracasei. According to high-efficiency anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) analysis, the rapid hydrolyzation of higher degree of polymerization (DP) fructans resulted in a transient increase in DP 3 content after 24 h of fermentation. The fructan and FODMAPs content of steamed bread with sourdough fermented by L. paracasei FJSSZ3L1 were 0.19 ± 0.01 g/100 g and 0.31 ± 0.01 g/100 g, respectively, which are significantly lower than the critical intake standards for IBS patients.

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Nutritional profile of rodent diets impacts experimental reproducibility in microbiome preclinical research.

Tuck, C. J., De Palma, G., Takami, K., Brant, B., Caminero, A., Reed, D. E., Muir, J. G., Gibson, P. R., Winterborn, A., Verdu, E. F., Bercik, P. & Vanner, S. (2020). Scientific Reports, 10(1), 1-13.

The lack of reproducibility of animal experimental results between laboratories, particularly in studies investigating the microbiota, has raised concern among the scientific community. Factors such as environment, stress and sex have been identified as contributors, whereas dietary composition has received less attention. This study firstly evaluated the use of commercially available rodent diets across research institutions, with 28 different diets reported by 45 survey respondents. Secondly, highly variable ingredient, FODMAP (Fermentable Oligo-, Di-, Mono-saccharides And Polyols) and gluten content was found between different commercially available rodent diets. Finally, 40 mice were randomized to four groups, each receiving a different commercially available rodent diet, and the dietary impact on cecal microbiota, short- and branched-chain fatty acid profiles was evaluated. The gut microbiota composition differed significantly between diets and sexes, with significantly different clusters in β-diversity. Total BCFA were highest (p = 0.01) and SCFA were lowest (p = 0.03) in mice fed a diet lower in FODMAPs and gluten. These results suggest that nutritional composition of commercially available rodent diets impact gut microbiota profiles and fermentation patterns, with major implications for the reproducibility of results across laboratories. However, further studies are required to elucidate the specific dietary factors driving these changes.

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Safety Information
Symbol : GHS05, GHS08
Signal Word : Danger
Hazard Statements : H314, H360
Precautionary Statements : P201, P202, P260, P264, P280
Safety Data Sheet
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