Chapter 1: Introduction
Chapter 2: Theory of the Analytical Procedure
Chapter 3: Test Booklet and Reagents
Chapter 4: Reagent Preparation
Chapter 5: Weighing of Samples
Chapter 6: Hydrolysis of Damaged Starch
Chapter 7: Hydrolysis of Dextrins to Glucose
Chapter 8: Glucose Determination (GOPOD Reagent)
Chapter 9: Calculation Simplified using MegaCalc
200 assays per kit
Prices exclude VAT
Available for shipping
|Content:||200 assays per kit|
Short term stability: 2-8oC,
Long term stability: See individual component labels
|Stability:||> 2 years under recommended storage conditions|
|Limit of Detection:||0.5 g/100 g|
|Total Assay Time:||~ 40 min|
|Application examples:||Cereal flours and other materials.|
|Method recognition:||AACC Method 76-31.01, ICC Standard No. 164 and RACI Standard Method|
The Starch Damage test kit is suitable for the measurement and analysis of starch damage in cereal flours.
See more of our starch assay kits.
- 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
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.Hide Abstract
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%.Hide Abstract
Gibson, T. S., Kaldor, C. J. & McCleary, B. V. (1993). Cereal Chemistry, 70(1), 47-51.
A commercially available enzymatic assay kit for the measurement of starch damage in wheat flour was compared with current standard methods, and the kit's 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%).Hide Abstract
Gibson, T. S., Al Qalla, H. & McCleary, B. V. (1992). Journal of Cereal Science, 15(1), 15-27.
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.Hide Abstract
Ai, Y., Gong, L., Reed, M., Huang, J., Zhang, Y. & Jane, J. L. (2016). Starch‐Stärke, 68(1-2), 131-139.
This study aimed to characterize structures, physical properties, and enzymatic-hydrolysis rate of starch isolated from bamboo seeds (Phyllostachys heterocycla var. pubescens (Mazel) Ohwi) in comparison with that of indica and japonica-rice starch. The bamboo seeds consisted of 68.2% starch (dry basis, db), which was lower than the indica and japonica-rice kernels (85.1 and 87.1%, respectively). The bamboo-seed starch also had compound granules, with morphology and sizes comparable to the rice starches. The bamboo-seed amylopectin had a similar branch-chain-length distribution to the indica-rice amylopectin, and both had longer average branch-chain length (DP = 19.1 and 19.7, respectively) than the japonica-rice amylopectin (DP = 17.1). The bamboo-seed and indica-rice starches showed higher onset gelatinization temperatures (T0 = 68.9 and 71.9°C, respectively) and gelatinization enthalpy changes (ΔH = 14.2 and 15.3 J/g), larger percentages of retrogradation (57.1 and 55.4%), but slower enzymatic-hydrolysis rates (50.6 and 46.3% at 24 h) than the japonica-rice starch (56.4°C, 14.0 J/g, 11.4%, and 66.2%, respectively). The bamboo-seed starch displayed a higher pasting temperature but substantially lower viscosity than the rice starches, which could be attributed to its larger amylose content and smaller molecular weight and gyration-radius of the amylopectin.Hide Abstract
Mancebo, C. M., Picón, J. & Gómez, M. (2015). LWT-Food Science and Technology, 64(1), 264-269.
The three main ingredients of cookies are wheat flour, fat and sugar. In gluten-free cookies the wheat flour must be replaced by other ingredients. The objective of the present study was to determine the effects of the flour properties of different gluten-free flours on cookie quality. A variety of different gluten-free flours, including yellow maize, precooked yellow maize, buckwheat, teff and short-grain and long-grain rice flour, were employed. The flour characteristics (protein, damaged starch content, particle size, flour hydration properties and oil absorption), dough properties (texture) and cookie parameters (final diameter, spread factor, texture, colour and acceptability) were evaluated. Coarse-grained rice flours produced cookies with a larger diameter and spread factor, darker colour and lower hardness. The rest of the gluten-free cookies had a lower spread ratio and greater hardness than wheat cookies, regardless of flour particle size. We have thus established that it is possible to obtain gluten-free cookies with organoleptic acceptability similar to that of cookies made from wheat flour. Nevertheless, cookie acceptability was hardly influenced by the cereal origin and their taste.Hide Abstract
Hossen, M. S., Sotome, I., Nanayama, K., Sasaki, T. & Okadome, H. (2015). Cereal Chemistry, 93(1), 53-57.
The digestibility and hydration properties of wet-ground submicron-scale rice flour were compared with those of dry-ground coarser microscale flours. The submicron flour (mean size 0.6 µm) was produced in a wet-media mill with 0.3 mm zirconia beads by continuous 24 h pulverization. The solubility, water absorption index, and swelling power increased as the mean particle size decreased, reaching maximum values in the submicron flour. Starch damage was high in the submicron flour, with the absence of intact starch granules. The digestibility also increased as the particle size decreased, and it was highest in the submicron flour. These results show that wet-ground submicron rice flour has different functional properties from dry-ground coarser flour. The digestibility was more strongly influenced by starch damage and the water absorption index than by the mean particle size.Hide Abstract
ShuJun, W., QuanWei, X., Wei, H., MaoMao, M. & Shuo, W. (2015). Journal of Food Safety and Quality, 6(5), 1783-1789.
Objective: To study the relationship between acrylamide levels produced in the Maillard reaction and the contents of damaged starch and reducing sugars. Methods: The grains of 3 wheat varieties were milled by Brabender mill and Buhler mill to produce 8 kinds of wheat flour samples. The content of damaged starch and reducing sugars were determined by Megazyme damaged starch assay kit and reducing sugar assay kit. The content of acrylamide in baked doughs was measured by high performance liquid chromatography. Result: There were significant differences in the damaged starch content of 8 kinds of wheat flours, ranging from 3.59% to 8.93%. Reducing sugar contents were in the range of 0.07%-1.1%. The acrylamide levels were in the range of 0.84-1.84 µg/mL in the baked doughs with yeast, while baked doughs without yeast presented the acrylamide levels in the range of 0.47-1.90 µg/mL. Conclusion: There was a positive correlation between damaged starch content and reducing sugar content. Inconsistent effect of damaged starch content and addition of yeast on the formation of acrylamide was noticed.Hide Abstract
Liu, C., Wang, S., Copeland, L. & Wang, S. (2015). LWT-Food Science and Technology, 64(2), 829-836.
The physicochemical and in vitro digestibility properties of starches isolated from four varieties of field peas grown in China were determined. Field pea starch granules were oval, round, even though there were many irregularly shaped granules with sizes between 21.5 and 23.9 µm. Zhongwan-6 showed the highest amylose content (36.0%), whereas Baofeng-3 showed the lowest amylose content (33.2%). All starches exhibited a C-type X-ray diffraction pattern, with relative crystallinity ranging between 27.0 and 30.2%. Zhongwan-6 had the highest relative crystallinity and the value of 1047/1022 cm-1. Baofeng-3 exhibited the highest swelling power (10.30 g/g) and Cai exhibited the lowest swelling power (8.45 g/g). Ma had the highest T0 and Tp, but the lowest ΔH and narrowest R, whereas Zhongwan-6 showed the lowest T0 and Tp, but the highest ΔH. Zhongwan-6 and Ma starches showed the lowest and highest pasting viscosities. The gel of Zhongwan-6 was the hardest and Baofeng-3 was the softest. The in vitro digestibility studies showed that Baofeng-3 starch may be more effective than other starches in moderating post-prandial blood glucose levels.Hide Abstract
Ramirez‐Cortes, R., Bello‐Pérez, L. A., Gonzalez‐Soto, R. A., Gutierrez‐Meraz, F. & Alvarez‐Ramirez, J. (2015). Starch‐Stärke, 68(5-6), 488-495.
Isolating starch from botanical sources has been widely reported on a small laboratory scale for characterization. However, isolation processes on a large laboratory scale intended for up-scaling have rarely been studied. In general, the laboratory-scale processes studied are batch processes, which limit the production rate for intensive commercialization. This work studied a large, laboratory-scale process for isolating starch from plantain fruits. The process is simple and involves a series of inclined sieves for separating solids and liquids. Hard green (unripe) pre-climacteric plantains (Musa paradisiaca L.) were collected and divided into four lots to evaluate the process. Analyses based on SEM, particle size distribution, X-ray diffraction, and thermal properties showed the reproducibility of the process, which exhibited an acceptable isolation yield (approximately 50%), purity (approximately 90%), and moisture content (approximately 5%). Moreover, the amount of damaged starch was not higher than 1.53/100 g.Hide Abstract
Trappey, E. F., Khouryieh, H., Aramouni, F. & Herald, T. (2014). Food Science and Technology International, 1082013214523632.
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 < 0.05) in the composition of sorghum flours of varying extraction rate, most notably for fiber and total starch contents. Flour particle size and starch damage were significantly impacted by extraction rate and speed of pin-milling. Water absorption increased significantly with increasing extraction rate and pin-milling speed. Breads produced from 60% extraction flour had significantly higher specific volumes, better crumb properties, and lower crumb firmness when compared with all other extractions and flour types. The specific volume of bread slices ranged from 2.01 mL/g (100% extraction, no pin-milling) to 2.54 mL/g (60% extraction, low-speed pin-milling), whereas the firmness ranged from 553.28 g (60% extraction, high-speed pin-milling) to 1096.26 g (commercial flour, no pin-milling). The bread characteristics were significantly impacted by flour properties, specifically particle size, starch damage, and fiber content (P < 0.05).Hide Abstract
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.
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.Hide Abstract
Hager, A. S., Czerny, M., Bez, J., Zannini, E. & Arendt, E. K. (2013). Journal of Cereal Science, 58(1), 156-163.
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.Hide Abstract
Cruz, B. R., Abraão, A. S., Lemos, A. M. & Nunes, F. M. (2013). Carbohydrate Polymers, 94(1), 594-602.
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.Hide Abstract
Lamsal, B. P. & Johnson, L. A. (2012). Journal of Cereal Science, 56(2), 253-259.
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.Hide Abstract
Hager, A. S., Wolter, A., Jacob, F., Zannini, E. & Arendt, E. K. (2012). Journal of Cereal Science, 56(2), 239-247.
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.Hide Abstract
Butterworth, P. J., Warren, F. J., Grassby, T., Patel, H. & Ellis, P. R. (2012). Carbohydrate Polymers, 87(3), 2189-2197.
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.Hide Abstract
Lamsal, B. P., Wang, H. & Johnson, L. A. (2011). Bioresource Technology, 102(12), 6680-6686.
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.Hide Abstract
Yao, N. & Seetharaman, K. (2010). International Journal of Food Science & Technology, 45(10), 2052-2061.
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.Hide Abstract
Himmelsbach, D. S., Manful, J. T. & Coker, R. D. (2008). Cereal chemistry, 85(3), 384-390.
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 Vh/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.Hide Abstract