Fructan Assay Kit

The Fructan Assay Kit is suitable for the specific measurement of fructan in plant extracts, animal feed and food products containing starch, sucrose and other sugars. It is used in three validated methods for the determination of fructan: AOAC method 999.03 (foods), AOAC method 2018.07 (Animal Feed) and AOAC method 2016.14 (infant formula and adult nutritionals).

New, improved procedure.

In the most recent development, a recombinant endo-levanase has been incorporated into the fructanase mixture, extending the use of the method to the measurement of levan-type fructans as are present in grasses such as timothy, cocksfoot, ryegrass and red fescue.

The method described in this booklet employs ultra-pure, recombinant enzymes and specifically measures fructans including inulin-type fructans from chicory, dahlia, jerusalem artichoke; highly branched fructans from onion and wheat stems and leaves; and levan-type fructans from pasture grasses such as timothy grass. The enzymes employed are completely devoid of contaminating enzymes active on β-glucan or gluco-oligosaccharides.


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Product Code
100 assays per kit

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Validation of Methods

AACC Method 32-32.01


AOAC Method 999.03
AOAC Method 2016.14
AOAC Method 2018.07


CODEX Method Type III 

Colourimetric method for the determination of Fructan in plant products, foods, infant formula, animal feed and other materials

(1) Sucrose + H2O → D-glucose + D-fructose

                             (β-amylase + maltase + pullulanase)
(2) Starch + maltosaccharides + H2O → D-glucose

(3) D-Glucose + D-fructose → D-sorbitol + D-mannitol

        (exo-inulinase + endo-inulinase + endo-levanase)
(4) Fructan + H2O → D-glucose + D-fructose

                                                    (100°C, 6 min)
(5) D-Glucose + D-fructose + PAHBAH → PAHBAH colour complex

Kit size:                             100 assays
Method:                             Spectrophotometric at 410 nm
Total assay time:              ~ 90 min
Detection limit:                 1-100% of sample weight
Application examples:
Flours, infant formula, animal feed, pet foods, plant materials (e.g. onion), food products and other materials
Method recognition:    
AOAC (Methods 999.03, 2016.14, 2018.07), AACC (Method 32-32.01) and CODEX (Type III Method)


  • Very cost effective
  • All kit reagents stable for > 2 years after preparation
  • Unaffected by high sucrose / reducing sugar concentrations
  • Fructan kits are only available from Megazyme
  • Simple format
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing
  • Standard included

Determination of Fructan (Inulin, FOS, Levan, and Branched Fructan) in Animal Food (Animal Feed, Pet Food, and Ingredients): Single-Laboratory Validation, First Action 2018.07. (2019).

McCleary, B. V., Charmier, L. M. J., McKie, V. A., Ciara McLoughlin, C. & Rogowski, A. (2019). Journal of AOAC International, 102(3), 2019 883.

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.

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.

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.

Physical, microscopic and chemical characterisation of industrial rye and wheat brans from the Nordic countries.

Kamal-Eldin, A., Lærke, H. N., Knudsen, K. E. B., Lampi, A. M., Piironen, V., Adlercreutz, H., Katina, K., Poutanen, K. & Aman, P. (2009). Food & Nutrition Research, 53.

Waxy endosperm accompanies increased fat and saccharide contents in bread wheat (Triticum aestivum) grain.

Yasui, T. & Ashida, K. (2011). Journal of Cereal Science, 53(1), 104-111.

Characterization and in vitro immunomodulatory screening of fructo-oligosaccharides of Asparagus racemosus Willd.

Thakur, M., Connellan, P., Deseo, M. A., Morris, C., Praznik, W., Loeppert, R. & Dixit, V. K. (2012). International Journal of Biological Macromolecules, 50(1), 77-81.

Steam‐girdling of barley (Hordeum vulgare) leaves leads to carbohydrate accumulation and accelerated leaf senescence, facilitating transcriptomic analysis of senescence‐associated genes.

Parrott, D. L., McInnerney, K., Feller, U. & Fischer, A. M. (2007). New Phytologist, 176(1), 56-69.

Contents of dietary fibre components and their relation to associated bioactive components in whole grain wheat samples from the HEALTHGRAIN diversity Screen.

Andersson, A. A. M., Andersson, R., Piironen, V., Lampi, A. M., Nyström, L., Boros, D., Fraś, A., Gebruers, K., Courtin, C. M., Delcour, J. A., Rakszegi, M., Bedo, Z., Ward, J. L., Shewry, P. R. & man, P. (2013). Food Chemistry, 136(3-4), 1243-1248.

Distribution and characterisation of fructan in wheat milling fractions.

Haskå, L., Nyman, M. & Andersson, R. (2008). Journal of Cereal Science, 48(3), 768-774.

Comparison of a colorimetric and a high‐performance liquid chromatography method for the determination of fructan in pasture grasses for horses.

Longland, A. C., Dhanoa, M. S. & Harris, P. A. (2012). Journal of the Science of Food and Agriculture, 92(9), 1878-1885.

Relationship of Grain Fructan Content to Degree of Polymerisation in Different Barleys.

Nemeth, C., Andersson, A. A. M., Andersson, R., Mangelsen, E., Sun, C. & Åman, P. (2014). Food and Nutrition Sciences, 2014, 5(6), 581-589.

Chain length of inulin affects its degradation and the microbiota in the gastrointestinal tract of weaned piglets after a short-term dietary application.

Paßlack, N., Al-Samman, M., Vahjen, W., Männer, K. & Zentek, J. (2012). Livestock Science, 149(1-2), 128-136.

How does the preparation of rye porridge affect molecular weight distribution of extractable dietary fibers?

Rakha, A., Åman, P. & Andersson, R. (2011). International Journal of Molecular Sciences, 12(5), 3381-3393.


R. S. & Singh, R. P. Singh. (2017). “Current Developments in Biotechnology and Bioengineering”, pp 423-446.

Processing effects on four prebiotic carbohydrates supplemented in an extruded cereal and a low pH drink.

Duar, R. M., Ang, P. T., Hoffman, M., Wehling, R., Hutkins, R. & Schlegel, V. (2015). Cogent Food & Agriculture, 1(1), 1013782.

Yacon (Smallanthus sonchifolius) and Lactobacillus acidophilus CRL 1014 reduce the early phases of colon carcinogenesis in male Wistar rats.

da Silva Almeida, A. P., Avi, C. M., Barbisan, L. F., de Moura, N. A., Caetano, B. F. R., Romualdo, G. R. & Sivieri, K. (2015). Food Research International, 74, 48-54.

Selected Sri Lankan food plants and other herbs as potential sources of inulin-type fructans.

Mudannayake, D. C., Wimalasiri, K. M. S., Silva, K. F. S. T. & Ajlouni, S. (2015). Journal of the National Science Foundation of Sri Lanka, 43(1).

Response surface methodology analysis of rice starch and fructo-oligosaccharides as substitutes for phosphate and dextrose in whole muscle cooked hams.

Resconi, V. C., Keenan, D. F., Gough, S., Doran, L., Allen, P., Kerry, J. P. & Hamill, R. M. (2015). L

Positive impact of a functional ingredient on hunger and satiety after ingestion of two meals with different characteristics.

Giuntini, E. B., Dan, M. C., Lui, M. C. Y., Lajolo, F. M. & Menezes, E. W. (2015). Food Research International, 76(3), 395-401.

Wheat grain filling is limited by grain filling capacity rather than the duration of flag leaf photosynthesis: a case study using NAM RNAi plants.

Borrill, P., Fahy, B., Smith, A. M. & Uauy, C. (2015). PloS One, 10(8), e0134947.
To choose a chapter, play the video and select the required chapter from the options on the video display.

Chapter 1: Introduction: Theory of the Analytical Procedure
Chapter 2: Kit Contents
Chapter 3: Preparation of Kit Reagents
Chapter 4: Preparation of Buffer Solutions
Chapter 5: Preparation of PAHBAH Reagent
Chapter 6: Preparation of Alkaline Borohydride Solution
Chapter 7: Weighing Samples
Chapter 8: Assay Procedure: Extraction & Filtration
Chapter 9: Assay Procedure: Removal of Sucrose, Starch & Reducing Sugars
Chapter 10: Assay Procedure: Hydrolysis & Measurement of Fructan
Chapter 11: Assay Procedure: Development of Colour with PAHBAH Reagent
Chapter 12: Calculations

Below you will find a link to our dedicated frequently asked questions section. Within this section you will find common questions and answers on a range of topics about the product.