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Megazyme/Total Dietary Fiber Controls/K-TDFC/Sufficient for 6 Controls

For use with the Total Dietary Fiber Assay Kit. Contains barley β-glucan, high amylose maize starch, wheat starch, casein, pectin and larch galactan. Wheat arabinoxylan is available on request.

View Megazyme’s latest Guide for Dietary Fiber Analysis.

Measuring dietary fibre.

McCleary, B. V. (1999). The World of Ingredients, 50-53. Link to Article Read Abstract Interest in dietary fibre is undergoing a dramatic revival thanks in part to the introduction of new carbohydrates as dietary fibre components. Much emphasis is being placed on determining how much fibre is present in a food. Linking a particular amount of fibre to a specific health benefit is now an important area of research. Total Dietary Fibre. The term \"dietary fibre” first appeared in 1953 and referred to hemicelluloses, celluloses and lignin (1). In 1974, Trowell (2) recommended this term as a replacement for the no longer acceptable term \"crude fibre” Burkitt (3) has likened the interest in dietary fibre to the growth of a river from its first trickle to a mighty torrent. He observes that dietary fibre \"was viewed as merely the less digestible constituent of food which exerts a laxative action by irritating the gut \"thus acquiring the designation \"roughage” a term which was later replaced by \"crude fibre” and ultimately by \"dietary fibre” Various definitions of dietary fibre have appeared over the years, partly due the various concepts used in deriving the term (i.e. origin of material, resistance to digestion, fermentation in the colon etc.), and partly to the difficulties associated with its measurement and labelling (4). The principle components of dietary fibre, as traditionally understood, are non-starch polysaccharides, which in plant fibre are principally hemicelluloses and celluloses, and the non-carbohydrate phenolic components, cutin, suberin and waxes with which they are associated in Nature.

Two issues in dietary fiber measurement.

McCleary, B. V. (2001). Cereal Foods World, 46(4), 164-165. Link to Article Read Abstract Enzyme activity and purity of these topics, the easiest to deal with is the importance of enzyme purity and activity. As a scientist actively involved in polysaccharide research over the past 25 years, I have come to appreciate the importance of enzyme purity and specificity in polysaccharide modification and measurement (7). These factors translate directly to dietary fiber (DF) methodology, because the major components of DF are carbohydrate polymers and oligomers. The committee report published in the March issue of Cereal FOODS WORLD refers only to the methodology for measuring enzyme purity and activity (8) that led up the AOAC method 985.29 (2). In this work enzyme purity was gauged by the lack of hydrolysis (i.e., complete recovery) of a particular DF component (e.g. β-glucan, larch galactan or citrus pectin). Enzyme activity was measured by the ability to completely hydrolyze representative starch and protein (namely wheat starch and casein). These requirements and restrictions on enzyme purity and activity were adequate at the time the method was initially developed and served as a useful working guide. However, it was recognized that there was a need for more stringent quality definitions and assay procedures for enzymes used in DF measurements.

Dietary fibre analysis.

McCleary, B. V. (2003). Proceedings of the Nutrition Society, 62, 3-9. Link to Article Read Abstract The gold standard method for the measurement of total dietary fibre is that of the Association of Official Analytical Chemists (2000; method 985.29). This procedure has been modified to allow measurement of soluble and insoluble dietary fibre, and buffers employed have been improved. However, the recognition of the fact that non-digestible oligosaccharides and resistant starch also behave physiologically as dietary fibre has necessitated a re-examination of the definition of dietary fibre, and in turn, a re-evaluation of the dietary fibre methods of the Association of Official Analytical Chemists. With this realisation, the American Association of Cereal Chemists appointed a scientific review committee and charged it with the task of reviewing and, if necessary, updating the definition of dietary fibre. It organised various workshops and accepted comments from interested parties worldwide through an interactive website. More recently, the (US) Food and Nutrition Board of the Institute of Health, National Academy of Sciences, under the oversight of the Standing Committee on the Scientific Evaluation of Dietary Reference Intakes, assembled a panel to develop a proposed definition(s) of dietary fibre. Various elements of these definitions were in agreement, but not all. What was clear from both reviews is that there is an immediate need to re-evaluate the methods that are used for dietary fibre measurement and to make appropriate changes where required, and to find new methods to fill gaps. In this presentation, the state of the art in measurement of total dietary fibre and dietary fibre components will be described and discussed, together with suggestions for future research.

Measurement of novel dietary fibres.

McCleary, B. V. & Rossiter, P. (2004). Journal of AOAC International, 87(3), 707-717. Link to Article Read Abstract With the recognition that resistant starch (RS) and nondigestible oligosaccharides (NDO) act physiologically as dietary fiber (DF), a need has developed for specific and reliable assay procedures for these components. The ability of AOAC DF methods to accurately measure RS is dependent on the nature of the RS being analyzed. In general, NDO are not measured at all by AOAC DF Methods 985.29 or 991.43, the one exception being the high molecular weight fraction of fructo-oligosaccharides. Values obtained for RS, in general, are not in good agreement with values obtained by in vitro procedures that more closely imitate the in vivo situation in the human digestive tract. Consequently, specific methods for the accurate measurement of RS and NDO have been developed and validated through interlaboratory studies. In this paper, modifications to AOAC fructan Method 999.03 to allow accurate measurement of enzymically produced fructo-oligosaccharides are described. Suggested modifications to AOAC DF methods to ensure complete removal of fructan and RS, and to simplify pH adjustment before amyloglucosidase addition, are also described.

An integrated procedure for the measurement of total dietary fibre (including resistant starch), non-digestible oligosaccharides and available carbohydrates.

McCleary, B. V. (2007). Analytical and Bioanalytical Chemistry, 389(1), 291-308. Link to Article Read Abstract A method is described for the measurement of dietary fibre, including resistant starch (RS), non-digestible oligosaccharides (NDO) and available carbohydrates. Basically, the sample is incubated with pancreatic α-amylase and amyloglucosidase under conditions very similar to those described in AOAC Official Method 2002.02 (RS). Reaction is terminated and high molecular weight resistant polysaccharides are precipitated from solution with alcohol and recovered by filtration. Recovery of RS (for most RS sources) is in line with published data from ileostomy studies. The aqueous ethanol extract is concentrated, desalted and analysed for NDO by high-performance liquid chromatography by a method similar to that described by Okuma (AOAC Method 2001.03), except that for logistical reasons, D-sorbitol is used as the internal standard in place of glycerol. Available carbohydrates, defined as D-glucose, D-fructose, sucrose, the D-glucose component of lactose, maltodextrins and non-resistant starch, are measured as D-glucose plus D-fructose in the sample after hydrolysis of oligosaccharides with a mixture of sucrase/maltase plus β-galactosidase.

Development and evaluation of an integrated method for the measurement of total dietary fibre.

McCleary, B. V., Mills, C. & Draga, A. (2009). Quality Assurance and Safety of Crops & Foods, 1(4), 213–224. Link to Article Read Abstract An integrated total dietary fibre (TDF) method, consistent with the recently accepted CODEX definition of dietary fibre, has been developed. The CODEX Committee on Nutrition and Foods for Special Dietary Uses (CCNFSDU) has been deliberating for the past 8 years on a definition for dietary fibre that correctly reflects the current consensus thinking on what should be included in this definition. As this definition was evolving, it became evident to us that neither of the currently available methods for TDF (AOAC Official Methods 985.29 and 991.43), nor a combination of these and other methods, could meet these requirements. Consequently, we developed an integrated TDF procedure, based on the principals of AOAC Official Methods 2002.02, 991.43 and 2001.03, that is compliant with the new CODEX definition. This procedure quantitates high- and low-molecular weight dietary fibres as defined, giving an accurate estimate of resistant starch and non-digestible oligosaccharides also referred to as low-molecular weight soluble dietary fibre. In this paper, the method is discussed, modifications to the method to improve simplicity and reproducibility are described, and the results of the first rounds of interlaboratory evaluation are reported.

Determination of total dietary fiber (CODEX definition) by enzymatic-gravimetric method and liquid chromatography: collaborative study.

McCleary, B. V., DeVries, J. W., Rader, J. I., Cohen, G., Prosky, L., Mugford, D. C., Champ, M. & Okuma, K. (2010). Journal of AOAC International, 93(1), 221-233. Link to Article Read Abstract A method for the determination of total dietary fiber (TDF), as defined by the CODEX Alimentarius, was validated in foods. Based upon the principles of AOAC Official MethodsSM 985.29, 991.43, 2001.03, and 2002.02, the method quantitates high- and low-molecular-weight dietary fiber (HMWDF and LMWDF, respectively). In 2007, McCleary described a method of extended enzymatic digestion at 37°C to simulate human intestinal digestion followed by gravimetric isolation and quantitation of HMWDF and the use of LC to quantitate low-molecular-weight soluble dietary fiber (LMWSDF). The method thus quantitates the complete range of dietary fiber components from resistant starch (by utilizing the digestion conditions of AOAC Method 2002.02) to digestion resistant oligosaccharides (by incorporating the deionization and LC procedures of AOAC Method 2001.03). The method was evaluated through an AOAC collaborative study. Eighteen laboratories participated with 16 laboratories returning valid assay data for 16 test portions (eight blind duplicates) consisting of samples with a range of traditional dietary fiber, resistant starch, and nondigestible oligosaccharides. The dietary fiber content of the eight test pairs ranged from 11.57 to 47.83. Digestion of samples under the conditions of AOAC Method 2002.02 followed by the isolation and gravimetric procedures of AOAC Methods 985.29 and 991.43 results in quantitation of HMWDF. The filtrate from the quantitation of HMWDF is concentrated, deionized, concentrated again, and analyzed by LC to determine the LMWSDF, i.e., all nondigestible oligosaccharides of degree of polymerization 3. TDF is calculated as the sum of HMWDF and LMWSDF. Repeatability standard deviations (Sr) ranged from 0.41 to 1.43, and reproducibility standard deviations (SR) ranged from 1.18 to 5.44. These results are comparable to other official dietary fiber methods, and the method is recommended for adoption as Official First Action.

Determination of insoluble, soluble, and total dietary fiber (codex definition) by enzymatic-gravimetric method and liquid chromatography: Collaborative Study.

McCleary, B. V., DeVries, J. W., Rader, J. I., Cohen, G., Prosky, L., Mugford, D. C., Champ, M. & Okuma, K. (2012). Journal of AOAC International, 95(3), 824-844. Link to Article Read Abstract A method for the determination of insoluble (IDF), soluble (SDF), and total dietary fiber (TDF), as defined by the CODEX Alimentarius, was validated in foods. Based upon the principles of AOAC Official MethodsSM 985.29, 991.43, 2001.03, and 2002.02, the method quantitates water-insoluble and water-soluble dietary fiber. This method extends the capabilities of the previously adopted AOAC Official Method 2009.01, Total Dietary Fiber in Foods, Enzymatic-Gravimetric-Liquid Chromatographic Method, applicable to plant material, foods, and food ingredients consistent with CODEX Definition 2009, including naturally occurring, isolated, modified, and synthetic polymers meeting that definition. The method was evaluated through an AOAC/AACC collaborative study. Twenty-two laboratories participated, with 19 laboratories returning valid assay data for 16 test portions (eight blind duplicates) consisting of samples with a range of traditional dietary fiber, resistant starch, and nondigestible oligosaccharides. The dietary fiber content of the eight test pairs ranged from 10.45 to 29.90%. Digestion of samples under the conditions of AOAC 2002.02 followed by the isolation, fractionation, and gravimetric procedures of AOAC 985.29 (and its extensions 991.42 and 993.19) and 991.43 results in quantitation of IDF and soluble dietary fiber that precipitates (SDFP). The filtrate from the quantitation of water-alcohol-insoluble dietary fiber is concentrated, deionized, concentrated again, and analyzed by LC to determine the SDF that remains soluble (SDFS), i.e., all dietary fiber polymers of degree of polymerization = 3 and higher, consisting primarily, but not exclusively, of oligosaccharides. SDF is calculated as the sum of SDFP and SDFS. TDF is calculated as the sum of IDF and SDF. The within-laboratory variability, repeatability SD (Sr), for IDF ranged from 0.13 to 0.71, and the between-laboratory variability, reproducibility SD (sR), for IDF ranged from 0.42 to 2.24. The within-laboratory variability sr for SDF ranged from 0.28 to 1.03, and the between-laboratory variability sR for SDF ranged from 0.85 to 1.66. The within-laboratory variability sr for TDF ranged from 0.47 to 1.41, and the between-laboratory variability sR for TDF ranged from 0.95 to 3.14. This is comparable to other official and approved dietary fiber methods, and the method is recommended for adoption as Official First Action.

Measurement of total dietary fiber using AOAC method 2009.01 (AACC International approved method 32-45.01): Evaluation and updates.

McCleary, B. V., Sloane, N., Draga, A. & Lazewska, I. (2013). Cereal Chemistry, 90(4), 396-414. Link to Article Read Abstract The Codex Committee on Methods of Analysis and Sampling recently recommended 14 methods for measurement of dietary fiber, eight of these being type I methods. Of these type I methods, AACC International Approved Method 32-45.01 (AOAC method 2009.01) is the only procedure that measures all of the dietary fiber components as defined by Codex Alimentarius. Other methods such as the Prosky method (AACCI Approved Method 32-05.01) give similar analytical data for the high-molecular-weight dietary fiber contents of food and vegetable products low in resistant starch. In the current work, AACCI Approved Method 32-45.01 has been modified to allow accurate measurement of samples high in particular fructooligosaccharides: for example, fructotriose, which, in the HPLC system used, chromatographs at the same point as disaccharides, meaning that it is currently not included in the measurement. Incubation of the resistant oligosaccharides fraction with sucrase/β-galactosidase removes disaccharides that interfere with the quantitation of this fraction. The dietary fiber value for resistant starch type 4 (RS4), varies significantly with different analytical methods, with much lower values being obtained with AACCI Approved Method 32-45.01 than with 32-05.01. This difference results from the greater susceptibility of RS4 to hydrolysis by pancreatic α-amylase than by bacterial α-amylase, and also a greater susceptibility to hydrolysis at lower temperatures. On hydrolysis of samples high in starch in the assay format of AACCI Approved Method 32-45.01 (AOAC method 2009.01), resistant maltodextrins are produced. The major component is a heptasaccharide that is highly resistant to hydrolysis by most of the starch-degrading enzymes studied. However, it is hydrolyzed by the maltase/amyloglucosidase/isomaltase enzyme complex present in the brush border lining of the small intestine. As a consequence, AOAC methods 2009.01 and 2011.25 (AACCI Approved Methods 32-45.01 and 32-50.01, respectively) must be updated to include an additional incubation with amyloglucosidase to remove these oligosaccharides.

Modification to AOAC Official Methods 2009.01 and 2011.25 to allow for minor overestimation of low molecular weight soluble dietary fiber in samples containing starch.

McCleary, B. V. (2014). Journal of AOAC International, 97(3), 896-901. Link to Article Read Abstract AOAC Official Methods 2009.01 and 2011.25 have been modified to allow removal of resistant maltodextrins produced on hydrolysis of various starches by the combination of pancreatic α-amylase and amyloglucosidase (AMG) used in these assay procedures. The major resistant maltodextrin, 63,65-di-α-D-glucosyl maltopentaose, is highly resistant to hydrolysis by microbial α-glucosidases, isoamylase, pullulanase, pancreatic, bacterial and fungal α-amylase and AMG. However, this oligosaccharide is hydrolyzed by the mucosal α-glucosidase complex of the pig small intestine (which is similar to the human small intestine), and thus must be removed in the analytical procedure. Hydrolysis of these oligosaccharides has been by incubation with a high concentration of a purified AMG at 60°C. This incubation results in no hydrolysis or loss of other resistant oligosaccharides such as FOS, GOS, XOS, resistant maltodextrins (e.g., Fibersol 2) or polydextrose. The effect of this additional incubation with AMG on the measured level of low molecular weight soluble dietary fiber (SDFS) and of total dietary fiber in a broad range of samples is reported. Results from this study demonstrate that the proposed modification can be used with confidence in the measurement of dietary fiber.

In vitro fermentation of spent turmeric powder with a mixed culture of pig faecal bacteria.

Han, K. H., Azuma, S. & Fukushima, M. (2014). Food & Function, 10, 2446-2452. Link to Article Read Abstract The fermentation potential of spent turmeric was studied in in vitro swine faecal batch cultures. The spent turmeric residue (the enzyme-resistant fraction from spent turmeric, EST) was obtained through the use of the digestive enzymes amyloglucosidase and pancreatin and compared to cellulose and high-amylose starch (HAS) as carbon sources. EST showed significant increases in total anaerobes, bifidobacteria, lactobacilli and lactic acid bacteria populations compared to cellulose at 12, 24 and 48 h, and the total anaerobic level in the HAS group was significantly higher than in the cellulose group at 24 and 48 h. However, a significant decrease in the coliform population was only found in the HAS group compared to the cellulose group at 48 h. The total short-chain fatty acid (SCFA) concentrations in the EST and HAS groups were significantly higher than that in the cellulose group at 12 h and 48h. However, there was no significant difference in the total SCFA concentration between the EST and HAS groups at 12 h and 48h. Ammonia and pH levels in the EST and HAS groups were significantly lower than those in the cellulose group at 24 and 48 h, but there was no significant difference between the EST and HAS groups. These results indicate that the fermentation potential of the enzyme-resistant fraction from spent turmeric is comparable to that of commercially established resistant starch.