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%.
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.
Sterilization in a liquid of a specific starch makes it slowly digestible in vitro and low glycemic in rats.
Severijnen, C., Abrahamse, E., Van der Beek, E. M., Buco, A., van de Heijning, B. J. M., van Laere, K. & Bouritius, H. (2007). The Journal of Nutrition, 137(10), 2202-2207.
Diabetics are recommended to eat a balanced diet containing normal amounts of carbohydrates, preferably those with a low glycemic index. For solid foods, this can be achieved by choosing whole-grain, fiber-rich products. For (sterilized) liquid products, such as meal replacers, the choices for carbohydrate sources are restricted due to technological limitations. Starches usually have a high glycemic index after sterilization in liquids, whereas low glycemic sugars and sugar replacers can only be used in limited amounts. Using an in vitro digestion assay, we identified a resistant starch (RS) source [modified high amylose starch (mHAS)] that might enable the production of a sterilized liquid product with a low glycemic index. Heating mHAS for 4–5 min in liquid increased the slowly digestible starch (SDS) fraction at the expense of the RS portion. The effect was temperature dependent and reached its maximum above 120°C. Heating at 130°C significantly reduced the RS fraction from 49 to 22%. The product remained stable for at least several months when stored at 4°C. To investigate whether a higher SDS fraction would result in a lower postprandial glycemic response, the sterilized mHAS solution was compared with rapidly digestible maltodextrin. Male Wistar rats received an i.g. bolus of 2.0 g available carbohydrate/kg body weight. Ingestion of heat-treated mHAS resulted in a significant attenuation of the postprandial plasma glucose and insulin responses compared with maltodextrin. mHAS appears to be a starch source which, after sterilization in a liquid product, acquires slow-release properties. The long-term stability of mHAS solutions indicates that this may provide a suitable carbohydrate source for low glycemic index liquid products for inclusion in a diabetes-specific diet.
Development of high amylose wheat through TILLING.
Slade, A. J., McGuire, C., Loeffler, D., Mullenberg, J., Skinner, W., Fazio, G., Holm, A., Brandt, K. M., Steine M. N., Goodstal, J. F. & Knauf, V. C. (2012). BMC Plant Biology, 12(1), 69.
Background: Wheat (Triticum spp.) is an important source of food worldwide and the focus of considerable efforts to identify new combinations of genetic diversity for crop improvement. In particular, wheat starch composition is a major target for changes that could benefit human health. Starches with increased levels of amylose are of interest because of the correlation between higher amylose content and elevated levels of resistant starch, which has been shown to have beneficial effects on health for combating obesity and diabetes. TILLING (Targeting Induced Local Lesions in Genomes) is a means to identify novel genetic variation without the need for direct selection of phenotypes. Results: Using TILLING to identify novel genetic variation in each of the A and B genomes in tetraploid durum wheat and the A, B and D genomes in hexaploid bread wheat, we have identified mutations in the form of single nucleotide polymorphisms (SNPs) in starch branching enzyme IIa genes (SBEIIa). Combining these new alleles of SBEIIa through breeding resulted in the development of high amylose durum and bread wheat varieties containing 47-55% amylose and having elevated resistant starch levels compared to wild-type wheat. High amylose lines also had reduced expression of SBEIIa RNA, changes in starch granule morphology and altered starch granule protein profiles as evaluated by mass spectrometry. Conclusions: We report the use of TILLING to develop new traits in crops with complex genomes without the use of transgenic modifications. Combined mutations in SBEIIa in durum and bread wheat varieties resulted in lines with significantly increased amylose and resistant starch contents.
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.
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.