The Starch Damage test kit is suitable for the measurement and analysis of starch damage in cereal flours.

An improved enzymic method for the measurement of starch damage in wheat flour.

Gibson, T. S., Al Qalla, H. & McCleary, B. V. (1992). Journal of Cereal Science, 15(1), 15-27. Link to Article Read Abstract An improved enzymic method for the determination of starch damage in wheat flour has been developed and characterized. The proposed method is simple and reliable, and enables up to 20 samples to be measured in duplicate in 2 h. A single assay takes approximately 40 min. The assay protocol is in two phases. In the first, the flour sample is incubated with purified fungal alpha-amylase to liberate damaged starch granules as soluble oligosaccharides. After centrifugation, the oligosaccharides in the supernatant are hydrolysed by amyloglucosidase to glucose in phase 2. The glucose is then quantified with a glucose oxidase/peroxidase reagent. The proposed method therefore avoids potential errors associated with existing standard assays, which employ unpurified amylase preparations and non-specific reducing group methods to quantify the hydrolytic products. Despite the use of purified assay components, the proposed starch damage method did not exhibit an absolute end-point to the action of alpha-amylase in phase 1. This was due to a low rate of hydrolysis of undamaged granules, and is a feature of enzymic methods for starch damage determination. Other amylolytic enzymes, including beta-amylase, isoamylase and pullulanase, and combinations of these enzymes, were evaluated as alternatives to alpha-amylase in the proposed method. These enzymes, when used alone, gave no benefits over the use of alpha-amylase. When used in combination with alpha-amylase, there was a synergistic action on undamaged granules. A test kit based on the assay format described in this paper is the subject of an international interlaboratory evaluation.

Collaborative evaluation of an enzymatic starch damage assay kit and comparison with other methods.

Gibson, T. S., Kaldor, C. J. & McCleary, B. V. (1993). Cereal Chem., 70(1), 47-51. Link to Article Read Abstract A commercially available enzymatic assay kit for the measurement of starch damage in wheat flour was compared with current standard methods, and the kits precision and repeatability were determined in a collaborative study. Starch damage values determined on a range of flours with the assay kit correlated well (r > 0.96) with those determined by existing standard enzymatic methods. The precision of the kit was evaluated in a comprehensive interlaboratory study. The kit procedure was found to be highly repeatable (relative standard deviation, 2.94-6.80%) and reproducible (relative standard deviation, 5.00-10.30%).

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. Link to Article Read Abstract 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. Link to Article Read Abstract 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.

Starch properties, in vitro digestibility and sensory evaluation of fresh egg pasta produced from oat, teff and wheat flour.

Hager, A. S., Czerny, M., Bez, J., Zannini, E. & Arendt, E. K. (2013). Journal of Cereal Science, 58(1), 156-163. Link to Article Read Abstract Specific dietary requirements, e.g. ceoliac disease, as well as increased consumer demand for products of high nutritional value, makes the production of pasta from alternative cereals interesting. Raw material characterisation showed that the utilisation of oat and teff flour is beneficial as these ingredients contain higher levels of fibre and mineral composition is superior to that of wheat. Starch properties significantly influence pasta quality and therefore damaged starch levels, amylase activity, pasting properties and gelatinisation temperatures of the flours were investigated. Fresh egg pasta based on wheat, oat and teff flour was produced. Sensory properties of oat spaghetti were found to be very close to that of wheat pasta but improvement of smoothness and aroma is necessary, while teff spaghetti showed reduced sensory quality. An in vitro enzymatic digestion was performed using a dialysis system to mimic the behaviour of pasta as eaten and make predictions on the glycemic index (GI). The predicted GI was highest for wheat pasta, followed by teff and oat. Ultra structure was studied using confocal laser scanning microscopy, allowing the visualisation of differences in starch granule size and shape as well as gelatinisation occurring during the cooking process.

Effect of sorghum flour composition and particle size on quality properties of gluten-free bread.

Trappey, E. F., Khouryieh, H., Aramouni, F. & Herald, T. (2014). Food Science and Technology International, 1082013214523632. Link to Article Read Abstract White, food-grade sorghum was milled to flour of varying extraction rates (60%, 80%, and 100%) and pin-milled at different speeds (no pin-milling, low-speed, and high-speed) to create flours of both variable composition and particle size. Flours were characterized for flour composition, total starch content, particle size distribution, color, damaged starch, and water absorption. Bread was characterized for specific volume, crumb structure properties, and crumb firmness. Significant differences were found (P P 

Nutritional properties and ultra-structure of commercial gluten free flours from different botanical sources compared to wheat flours.

Hager, A. S., Wolter, A., Jacob, F., Zannini, E. & Arendt, E. K. (2012). Journal of Cereal Science, 56(2), 239-247. Link to Article Read Abstract Coeliac patients suffer from an immune mediated disease, triggered by the ingestion of a protein composite (gluten) found in wheat, rye and barley. Consequently, there is a need for products such as bread or pasta, made from alternative cereal grains or pseudocereals. A fair proportion of the gluten free products currently on the market are nutritionally inadequate. Hence, it was the aim of this study to investigate the nutrient composition of seven commonly used commercial gluten free flours (oat, rice, sorghum, maize, teff, buckwheat and quinoa) and compare them to wheat and wholemeal wheat flour. In addition to the levels of all major compounds, also mineral composition, fatty acid profile, phytate, polyphenols and folate content were determined. Furthermore, properties of carbohydrates were studied in greater detail, looking at total and damaged starch levels; total, soluble and insoluble dietary fibre content as well as amylose/amylopectin ratio. Proteins were further investigated by means of capillary electrophoreses. Additionally, the ultra-structure of these materials was explored using scanning electron microscopy. The results show that maize and rice flour are poor regarding their nutritional value (low protein, fibre, folate contents). In contrast, teff as well as the pseudocereals quinoa and buckwheat show a favourable fatty acid composition and are high in protein and folate. In particular, quinoa and teff are characterised by high fibre content and are high in calcium, magnesium and iron. Therefore these flours represent nutrient dense raw materials for the production of gluten free foods.

Quality variations in flours used for pretzel manufacturing.

Yao, N. & Seetharaman, K. (2010). International Journal of Food Science & Technology, 45(10), 2052-2061. Link to Article Read Abstract Research on the flour properties and their influence on pretzel characteristics is scarce. In the first part of the study, flour protein quantity and quality, flour pasting properties and solvent retention properties of 108 flour samples were investigated to help profile the flour properties used by the pretzel industry. Four different flours with a wider protein range than what was revealed in the flour evaluation were selected to produce pretzels and to determine the relationship between flour properties and the final product quality. Pretzel hardness, colour and pasting properties were used as a measure of pretzel quality. Results indicated that hard wheat flour would produce a harder pretzel but would not affect the surface colour of final product. However, soft wheat flour with a lower damaged starch, low water absorption levels and lower water binding powers during operations is desired for making hard pretzel.

Effect of corn preparation methods on dry-grind ethanol production by granular starch hydrolysis and partitioning of spent beer solids.

Lamsal, B. P., Wang, H. & Johnson, L. A. (2011). Bioresource Technology, 102(12), 6680-6686. Link to Article Read Abstract Two corn preparation methods, rollermill flaking and hammermill grinding, were compared for efficient processing of corn into ethanol by granular starch hydrolysis and simultaneous fermentation by yeast Saccharomyces cerevisiae. Corn was either ground in a hammermill with different size screens or crushed in a smooth-surfaced rollermill at different roller gap settings. The partitioning of beer solids and size distribution of solids in the thin stillage were compared. The mean particle diameter d₅₀ for preparations varied with set-ups and ranged between 210 and 340 µm for ground corn, and 1180–1267 µm for flaked corn. The ethanol concentrations in beer were similar (18–19% v/v) for ground and flaked preparations, however, ethanol productivity increased with reduced particle size. Roller versus hammermilling of corn reduced solids in thin stillage by 28%, and doubled the volume percent of fines (d₅₀ 7 µm) in thin stillage and decreased coarse (d₅₀ 122 µm) by half compared to hammermilling.

Flaking as a corn preparation technique for dry-grind ethanol production using raw starch hydrolysis.

Lamsal, B. P. & Johnson, L. A. (2012). Journal of Cereal Science, 56(2), 253-259. Link to Article Read Abstract A 2³ full-factorial study was designed to study the effect of corn preparation methods (flaking and grinding) on dry-grind ethanol performance using raw starch hydrolysis (RSH) process. Moisture content (15, 22%), flaker roller gapsetting (0.508 mm, 1.016 mm), and grinding were studied. Fifteen hundred g of corn samples were cracked, roller pressed, and were either ground further or retained, along with control ground corn. A bimodal size distribution was observed for ground corn, regardless of flaking. Moisture at 22% resulted in bigger-sized flakes with d₅₀ between ~1.3 and 1.8 mm, compared to ~138–169 µm for ground corn. Not all ground corn resulted in higher ethanol concentration in fermentation beer; the ethanol levels in beer did not reflect the starch hydrolysis trend that favored ground corn. In a related study, the beer ethanol concentration did not show a clear trend with rollermill gapsetting while fermenting the flakes produced at 0.203, 0.305, 0.406, and 0.508 mm gapsettings. Generally, flakes from corn at 22% moisture resulted in higher ethanol content in beer. Rollermill flaking was found comparable to hammermill grinding for dry-grind corn ethanol via raw starch hydrolysis and yeast fermentation.

Chemical composition and functional properties of native chestnut starch (Castanea sativa Mill).

Cruz, B. R., Abraão, A. S., Lemos, A. M. & Nunes, F. M. (2013). Carbohydrate Polymers, 94(1), 594-602. Link to Article Read Abstract Starch isolation methods can change their physico-chemical and functional characteristics hindering the establishment of a starch-food functionality relation. A simple high yield and soft isolation method was applied for chestnut (Castanea sativa Mill) starch consisting in steeping and fruit disintegration in a 25 mM sodium bisulfite solution and purification by sedimentation. Starch integrity, physico-chemical composition, morphology and functional properties were determined, being observed significant differences from previous described methods for chestnut starch isolation. The X-ray pattern was of B-type, with a degree of crystallinity ranging from 51% to 9%, dependent on the starch moisture content. The onset, peak, and conclusion gelatinization temperatures were 57.1°C, 61.9°C and 67.9°C, respectively. Total amylose content was 26.6%, and there was not found any evidence for lipid complexed amylose. Swelling power at 90°C was 19 g/g starch, and the amount of leached amylose was 78% of the total amylose content. Native chestnut starch presents a type B pasting profile similar to corn starch but with a lower gelatinization (56.1°C) and peak viscosity (79.5°C) temperatures, making native chestnut starch a potential technological alternative to corn starch, especially in application where lower processing temperatures are needed.

Changes in rice with variable temperature parboiling: thermal and spectroscopic assessment.

Himmelsbach, D. S., Manful, J. T. & Coker, R. D. (2008). Cereal chemistry, 85(3), 384-390. Link to Article Read Abstract Rapid visco analysis (RVA) and differential scannning calorimetry (DSC) provided overall assessments of the effects of variable temperature soaking at 30, 50, 70, and 90°C and steaming at 4, 8, and 12 min. Calculation of the relative parboiling index (RPI) and percent gelatinization provided good metrics for determining the overall effects of partial parboiling. FT-Raman and solid-state ¹³C CP-MAS NMR spectroscopies provided insight to conformational changes in protein and starch of paddy rice under various parboiling conditions. RVA showed lower pasting curves and DSC showed lower ΔH with increased temperature and steaming times. A large decrease in viscosity occurred with only the 30-4 treatment as opposed to raw rice. This observation was consistent with FT-Raman results that indicated substantial conversion of the protein from α-helix to other conformations. DSC indicated incomplete gelatinization of starch, even with 90°C soaking and 12 min of steaming. Solid-state ¹³C CP-MAS NMR spectroscopy confirmed this result. However, it indicated the percent of V_h/amorphous plus the remaining crystalline starch in the 90-12 treatment was equal to the amorphous and partially-ordered starch in commercially parboiled rice. These results suggest that partial parboiling, 90°C soaking, and more than 8 min of steaming (ideally ≈12 min) of paddy rice is sufficient to induce changes that inactivate enzymes and provide enough starch gelatinization to prevent kernel breakage.

Determination of formulation and processing factors affecting slowly digestible starch, protein digestibility and antioxidant capacity of extruded sorghum–maize composite flour.

Licata, R., Chu, J., Wang, S., Coorey, R., James, A., Zhao, Y. & Johnson, S. (2014). International Journal of Food Science & Technology, 49(5), 1408-1419. Link to Article Read Abstract High-temperature high-pressure extrusion of sorghum–maize composite flour, of potential for healthy food manufacture, was investigated by factorial experimental design to determine the effect of level of sorghum in dry mix (15–60%); final barrel zone temperature (120–150°C); total moisture in barrel (21.4–25.8%); total input rate (2.3–6.8 kg h^-1); and screw speed (250–450 rpm) on extrudate slowly digestible starch (SDS), phenolic content, antioxidant capacity, protein digestibility, density and expansion ratio. Extrudate SDS increased with increasing sorghum level and decreased as the barrel temperature increased. Total phenolic content and antioxidant capacity were positively associated with sorghum level. Protein digestibility was associated negatively with sorghum level and positively with barrel temperature. Extrudate density was associated positively with total moisture and negatively with barrel temperature and input rate. Sorghum in dry mix, final barrel zone temperature and total moisture in barrel were the three most significant independent variables influencing extrudate dependant variables.

Analysis of starch amylolysis using plots for first-order kinetics.

Butterworth, P. J., Warren, F. J., Grassby, T., Patel, H. & Ellis, P. R. (2012). Carbohydrate Polymers, 87(3), 2189-2197. Link to Article Read Abstract Investigators often study product release from starches during prolonged incubations with α-amylase in vitro. The reaction time courses usually fit to a linear form of a first order rate equation, i.e., ln[(C∞ − Ct)/C∞] = −kt. This equation calls for an accurate estimate of C∞, i.e., the concentration of product at the end of the reaction. Estimates of C∞ from digestibility curves can be unreliable. The Guggenheim method does not require prior knowledge of C∞ but seems not to have been applied to starch hydrolysis data. An alternative method is also available in which the logarithm of the slope (LOS) of a digestibility curve at various time points is plotted against time. This allows estimations of both k and C∞ and can also reveal whether changes occur in digestion rate from rapid to slow as digestion proceeds. We describe the Guggenheim and LOS methods and provide examples of their application to starch digestibility data. Validation of Methods

AACC Method 76-31.01

ICC Standard No. 164

RACI Standard Method

Colourimetric method for the determination of Starch Damage in cereal floursPrinciple: (fungal α-amylase)(1) Damaged (or gelatinised) starch + H₂O → maltodextrins (amyloglucosidase)(2) Maltodextrins + H₂O → D-glucose (glucose oxidase)(3) D-Glucose + H₂O + O₂ → D-gluconate + H₂O₂ (peroxidase)(4) 2H₂O₂ + p-hydroxybenzoic acid + 4-aminoantipyrine → quinoneimine + 4H₂O

Kit size: 200 assaysMethod: Spectrophotometric at 510 nmTotal assay time:~ 40 minDetection limit:0.5-100% of sample weightApplication examples: Cereal flours and other materialsMethod recognition: AACC (Method 76-31.01), ICC (Standard No. 164) and RACI (Standard Method)

Advantages

• Very cost effective
• All reagents stable for > 2 years after preparation
• Only enzymatic kit available
• Very specific
• Simple format
• Mega-Calc™ software tool is available from our website for hassle-free raw data processing
• Standard included

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