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Starch Damage Assay Kit

Play Training Video

00:04  Introduction
01:24   Principle
02:22   Reagent Preparation
04:58   Weighing of Samples
05:51    Addition of α-amylase
07:07   Addition of amyloglucosidase
07:49   Glucose determination
09:16    Calculations
11:19      Further information

Starch Damage Assay Kit K-SDAM Scheme
Product code: K-SDAM

200 assays per kit

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Content: 200 assays per kit
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 1 year under recommended storage conditions
Analyte: Starch Damage
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 510
Signal Response: Increase
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 determination of starch damage in wheat flour / cereal flours.

The milling of wheat causes physical damage to a proportion of the starch granules of the flour. The level of starch damage directly affects water absorption and dough mixing properties of the flour and is thus of technological significance.

See more of our starch assay kits.

Scheme-K-SDAM SDAM Megazyme

  • 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
Validation of Methods
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance
Megazyme publication
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.

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Megazyme publication
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%.

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Megazyme publication

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 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%).

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Megazyme publication
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.

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.

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Comparison of hydration water properties of common and durum wheat brans upon grinding with different loading modes.

Barbar, R., Mayer-Laigle, C., Beaugrand, J., Cuq, B. & Barron, C. (2023). Journal of Cereal Science, 114, 103786.

Wheat bran brings healthy properties in food products. However, its incorporation requires a first milling step during which it is subject to various loading modes which have an influence on its properties. This study investigated the influence of the loading modes (high shear or impact) generated by grinders during the milling on the hydration properties of common and durum wheat brans. An original study at molecular scale to target the distribution and intensity of hydration water bonds was carried out by gravimetric and spectroscopic methods. Results were analyzed in regards to the particle size distribution and shape factors as well as biochemical composition to highlight the process-structure-function relationships at macro and microscales. Impact grinder has a stronger effect on water vapor sorption capacity and FTIR multimer water at 3600 cm−1 of durum wheat bran as well as a red shift in the band at 1030 cm−1 related to a high decrease in residual starch crystallinity degree (about 17%). High shear grinder tends to increase the proportion of water strongly bound. Low field NMR analysis revealed differences in the high mobility water peak and a significant lower relaxation time T2 for native and ground durum wheat bran (up to 1.7 fold less).

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Modification of Acorn Starch Structure and Properties by High Hydrostatic Pressure.

Castro, L. M., Caço, A. I., Pereira, C. F., Sousa, S. C., Brassesco, M. E., Machado, M., Ramos, O. L., Alexandre, E. M. C., Saraiva, J. A. & Pintado, M. (2023). Gels, 9(9), 757.

Despite being rich in starch, over half of acorn production is undervalued. High hydrostatic pressure was used to modify the properties of Q. pyrenaica (0.1 and 460 MPa for 20 min) and Q. robur (0.1 and 333 MPa for 17.4 min) acorn starches to obtain high-valued ingredients. Pressure significantly altered the span distribution and heterogeneity of the acorn starch granules depending on the species, but their morphology was unaffected. Pressurization increased the amylose/amylopectin ratio and damaged starch contents, but the effect was more prominent in Q. pyrenaica than in Q. robur. However, the polymorphism, relative crystallinity, gelatinization temperatures, and enthalpies were preserved. The pressure effect on the starch properties depended on the property and species. The solubility, swelling power, and acorn gels’ resistance towards deformation for both species decreased after pressurization. For Q. pyrenaica starch, the in vitro digestibility increased, but the pseudoplastic behavior decreased after pressurization. No differences were seen for Q. robur. Regarding the commercial starch, acorn starches had lower gelatinization temperatures and enthalpies, lower in vitro digestibility, lower resistance towards deformation, superior pseudoplastic behavior, and overall higher solubility and swelling power until 80 °C. This encourages the usage of acorn starches as a new food ingredient.

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The Physicochemical Properties of Starch from Tongil-type Rice Varieties.

Shin, J. H., Han, C. M., Song, Y. U., Kim, S. K. & Ryu, J. G. (2023). The Korean Journal of Crop Science, 68(3), 99-105.

Starch characteristics and pasting properties of Tongil-type rice varieties with different amylose content were analyzed. Three different Tongil-type rice varieties and one Japonica-type rice variety were examined for their properties. Tongil-type rice varieties have longer panicles and higher rice yield (721-765 kg per 10a) than Boramchan, a Japonica-type high-yield rice variety. The protein content of the Tongil-type rice variety was higher than that of Boramchan. Japonica-type rice varieties had lower amylose content than Japonica-type rice varieties, except for Amimyeon (Milyang355). Amimyeon had higher protein content than the other varieties, and its amylose content was particularly high at 39.2%. The distribution of starch granule sizes of the four varieties was similar, and the particle size corresponding to D50 was approximately 87.8-81.9 μm. The pasting properties of rice flour varied among varieties. The Dasanbyeo and Hanarum2 amylogram patterns were similar. These two varieties had a higher peak viscosity (PV) and lower setback (SB) than Boramchan. In the case of Amimyeon, the hot paste viscosity (HPV), cool paste viscosity (CPV), and SB were significantly higher than those of the other Toingil-type varieties, and the breakdown (BD) was very low, showing pasting properties significantly different from that of the other varieties. The onset (To), peak (Tp), and conclusion temperature (Tc) of gelatinization of rice flour from Amimyeon were lower than those of the other tested varieties. In addition, the gelatinization temperature, measured using differnetial scanning calorimetry (DSC), and BD, measured using al rapid visco analyzer (RVA) were low in Amimyeon. Amylose content showed a high positive correlation with pasting time (PTi), HPV, and SB and a negative correlation with PV and BD.

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Corn kernel hardness and drying temperature affect particle size post-hammer-milling and pellet quality in broiler and swine diets.

Cabañas-Ojeda, J. A., Mejia-Abaunza, N. J., Lozano-Cruz, P. A., Aragão-Netto, V., Brown, S., Rubio, A., Fahrenholz, A. & Oviedo-Rondón, E. O. (2023). Animal Feed Science and Technology, 304, 115744.

The particle size (PS) of ground corn affects the live performance, nutrient digestibility, energy utilization, and gut health of poultry and swine. Endosperm hardness influences kernel breakability. The interactive effects of corn moisture content at harvest and drying temperature (DT) can also affect starch-protein matrix structure, breakage susceptibility, and pellet durability. The objective of this study was to evaluate the effects of drying temperature (35 or 120 °C) in two yellow-dent corn hybrids with different kernel hardness (average and hard) on the geometric mean diameter (dgw) and standard deviation by weight (Sgw) when hammermilled, and pellet quality. Four different settings combinations of tip speeds (4213, 3862, 3335, and 2458 m/min) and screen of 8/64", 3.18 mm (8 −8); 20/64", 7.94 mm (20 −20); 24/64", 9.53 mm (24 −24); and 24/64", 9.53 mm (24 −24) respectively were used to target four dgw (400, 700, 925, and 1250 µm). After grinding, the 400 and 700 µm corn were included in broiler and sow diets to assess the impact of these treatments on the pellet durability index (PDI). Grinding and pelleting data were analyzed using a 2 × 2 factorial arrangement of two corn varieties (CV) and two DT in a randomized complete block design with blocks by grinding or pelleting event. The average hardness CV generally had the lowest dgw. The hard CV had the biggest dgw when kernels were dried at 35 °C, and the tip speed setting was 3335 m/min. Drying at 120 °C increased dgw for the average hardness CV when the target PS were 700, 925, and 1250 µm, but caused no increase in dgw at the lowest target PS. In contrast, the dgw of hard CV was not affected by DT when ground to target 700 and 1250 µm and increased at 400 µm with the high DT. The average hardness CV had higher Sgw, especially when dried at 35 °C. Higher particle uniformity (lower Sgw) was observed with hard CV. Diets containing the hard CV presented better PDI than diets with the average CV. Better broiler feed PDI was observed with corn dried at 120 °C than at 35 °C. However, this effect was the opposite when grinding at a lower PS target for sow gestation diets. The PS distribution and damaged starch of corn affected PDI. Higher corn DT affected dgw and Sgw differently depending on CV and improved PDI in broiler diets, while 35 °C DT improved PDI in sow diets.

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Identification and Characterization of Waxy Bread Wheat Carrying a Novel Wx-B1 Allele.

Sung, Y., Kim, K., Park, J., Kang, S., Park, C., Cho, S. & Kim, C. (2023), In Press.

Starch content in wheat is an important characteristic for various purposes. Starch is composed of two components known as amylose and amylopectin, and the proportion between these two components plays a crucial role in determining the properties and structure of starch. One of the key genes involved in starch biosynthesis in wheat is granule bound starch synthase I (GBSSI), also known as the waxy protein, which synthesizes amylose in the endosperm. A mutant cultivar, named Gunji-3, was created by crossing the waxy type wheat (Shinmichal 1) with the wild type bread wheat (Keumkang). When comparing the Wx-B1 allele of the waxy protein gene in Gunji-3 with that of the waxy type wheat, there were 12 SNPs and one deletion difference. Additionally, when comparing amino acid sequences with Wx-B1 alleles, differences at two positions are found, indicating that the mutant carries new Wx-B1 allele which is named as Wx-B1o. In terms of the physicochemical properties of Gunji-3 starch, it had a lower amylose content than the parental wheat varieties and showed higher protein content and greater swelling power. These newly introduced waxy type wheat could provide important basis not only for understanding the starch characteristics of wheat but also for various breeding programs.

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Impact of different preparation methods on the properties of brown rice flour and the cooking stability of brown rice noodles and the underlying mechanism: Microstructure, starch-protein distribution, moisture migration.

Sun, Z., Lyu, Q., Zhuang, K., Chen, L., Wang, G., Wang, Y., Wang, Y. & Ding, W. (2023). LWT, 181, 114697.

Brown rice flour (BRF) and corresponding brown rice noodle (BRN) were prepared by different methods and the mechanism of impact of extruded rice bran on cooking stability of BRN was investigated. The particle size distribution, damaged starch content and pasting properties of BRFs were evaluated and the texture properties, microstructure and moisture distribution during cooking of BRNs were investigated. The average particle size and damaged starch of brown rice flour refilled with extruded rice bran (BRF-E) and brown rice flour refilled with nonextruded rice bran (BRF-NE) were obviously less than that of the brown rice flour by fully crushed brown rice (BRF-F). The peak viscosity (PV), setback viscosity (SV) and pasting temperature (PT) of BRF-E and BRF-NE were significantly higher than that of BRF-F. The results of scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) showed that the microstructure of BRN-E was most compact, which indicated that using BRF refilled with extruded rice bran to produce BRN could promote BRN formed a compact texture structure. Meanwhile, water migration and texture properties analysis demonstrated that BRN-E had better cooking stability and chewiness.

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Changes on Techno-Functional, Thermal, Rheological, and Microstructural Properties of Tef Flours Induced by Microwave Radiation-Development of New Improved Gluten-Free Ingredients.

Calix-Rivera, C. S., Villanueva, M., Náthia-Neves, G. & Ronda, F. (2023). Foods, 12(6), 1345.

Tef [Eragrostis tef (Zucc.) Trotter] flour is a gluten-free cereal rich in fiber, minerals, vitamins, and antioxidants, which offers a promising alternative for new food development. This study investigated the effect of microwave radiation (MW) on the techno-functional, thermal, rheological and microstructural properties of tef flours. White and brown tef grains were milled and microwaved at different moisture contents (MC) (15%, 20% and 25%) for a total irradiation time of 480 s. The morphological structure of tef flours was affected by MW treatment, and its particle size and hydration properties increased after the treatment. Lower peak, breakdown, and setback viscosities, up to 45%, 96%, and 67% below those of the control (untreated) samples, and higher pasting temperature, up to 8 °C in the 25% MC samples, were observed. From FTIR analysis a disruption of short-range molecular order was concluded, while DSC confirmed an increased stability of starch crystallites. Rheological analysis of the gels made from the treated samples revealed that MW had a structuring and stabilizing effect on all samples, leading to higher viscoelastic moduli, G′ and G″, and the maximum stress the gels withstood before breaking their structure, τmax. The MC of the flours during the MWT drove the modification of the techno-functional properties of the tef flours and the gel rheological and thermal characteristics. These results suggest that MW-treated tef flours are potential ingredients for improving the technological, nutritional and sensory quality of food products.

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Applicability of Flours from Pigmented and Glutinous Rice in Gluten-Free Bread Baking.

Burešová, I., Červenka, L., Šebestíková, R., Augustová, M. & Jarošová, A. (2023). Foods, 12(6), 1324.

The flours from pigmented and glutinous rice have a great potential to increase the nutritional quality of gluten-free breads. The characteristics of whole-meal fine flours, slurries, doughs, and breads prepared from black, red, and white glutinous rice were, therefore, compared with commercially used refined fine and semi-coarse white rice flours. The pasting properties of different flours were strongly influenced by the type of rice they were made from. Slurries with red and glutinous flours exhibited a shift to a lower pasting temperature T0, lower values of ηPeak and ηFinal, as well as higher values of the breakdown and setback region than the slurries with refined flours. The slurry with black flour exhibited high values of viscosity η0 and breakdown, together with low values of ηFinal, setback region and total setback. Bread characteristics were strongly correlated with the pasting properties. The presence of bran particles did not negatively impact loaf volume, crumb hardness, springiness, and chewiness. Some defects were observed in glutinous and red rice bread. Fine flour exhibited better baking performance than semi-coarse flour. Glutinous flour has the potential to become an ingredient in gluten-free baking. The applicability of various black and red rice flours may be limited by the flavor and the taste.

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Exploring the nutritional potential, anti-nutritional components and carbohydrate fractions of Indian pigmented maize.

Gogoi, P., Sharma, P., Mahajan, A., Goudar, G., Kumar, A., Sreedhar, M., Singh, M. & Longvah, T. (2022). Food Chemistry Advances, 100176.

Seventeen pigmented maize cultivated in different locations of India were collected and evaluated for their nutritional, anti-nutritional and starch profile. A wide variation was observed in all the parameters evaluated. Protein and total dietary fiber content varied between 7.18 to 10.85 g/100g and 8.76 to 14.04 g/100g, respectively. Among the water-soluble vitamins, B2 varied from 0.071 to 0.134; B3, 1.50 to 2.79; B5, 0.17 to 0.70 and B6, 0.141 to 0.245 mg/100g; while total tocopherol and tocotrienol varied 1417 to 2246 µg/100g and 18.57 to 30.01 µg/100g, respectively. The resistant starch, total starch and amylose varied significantly and observed between 0.49 to 4.78 %, 61.33 to 69.65 % and 10.91 to 32.01 %, respectively. Among the minerals, Fe and Zn were observed between 1.51 to 5.01 and 2.25 to 5.51 mg/100g, respectively. Total polyunsaturated and total monounsaturated fatty acids were observed up to 45.72 and 55.54 %, respectively. Amongst the anti-nutrients, phytate was detected the lowest in Laal makki, whereas total and soluble oxalates were the lowest in Kashmiri red maize.

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Pasting and rheological properties of water caltrop starch as affected by the addition of konjac glucomannan, guar gum and xanthan gum.

Lan, Y. C. & Lai, L. S. (2022). Food Hydrocolloids, 108245.

Water caltrop starch (WCS) isolated from the dehydrated water caltrop (DWCS) showed higher content of protein and ash as compared to starch isolated from fresh water caltrop (FWCS), leading to slightly lower starch purity. However, the pasting profiles of DWCS and FWCS are similar under a constant solid content of 4% (w/w). WCS/gum mixed systems contain 0.3%–0.9% gum generally showed higher pasting temperature, peak viscosity, and final viscosity than the starch alone system. The increase in peak and final viscosity during rapid-viscosity analysis was most pronounced for WCS/gum mixed systems with konjac glucomannan (KG), followed by guar gum (GG) and xanthan gum (XG). All WCS/gum mixed systems showed shear thinning and thixotropic characteristics. The extent of pseudoplasticity and shear stability, as evidenced by the smaller flow behavior index (n) and area of the hysteresis loop, was most pronounced for WCS/gum mixed system with XG, followed by mixed systems with GG and KG. Dynamic viscoelastic analysis revealed that the addition of gum increased both the storage and loss moduli of WCS, and lowered the frequency dependence of the loss factor significantly. However, the loss factor of WCS/gum mixed systems was generally higher than that of the starch alone system, implying the addition of gums modified the rheological characteristic of WCS to more viscous dominant behavior.

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Effects of milling methods on the properties of rice flour and steamed rice cakes.

Wang, G., Yan, X., Wang, B., Hu, X., Chen, X. & Ding, W. (2022). LWT, 167, 113848.

To investigate the effects of milling methods on the properties of rice flour and steamed rice cakes (SRCs), dry, semi-dry, and wet milling methods were compared. Damaged starch, particle size distribution, water hydration properties, gelatinization properties, gel strength, thermal properties, glass transition temperature, dynamic rheological properties of rice flour, sensory and textural properties of SRCs were evaluated. Semidry-milling rice flour (SF) had much less damaged starch and smaller particle size compared with dry-milling rice flour (DF). The pasting, hydration, and rheological properties of SF were similar to that of wet-milling rice flour (WF). Except for adhesion and appearance, the sensory and textural properties of SF SRCs were significantly better than that of DF SRCs, while comparable to WF SRCs. The results showed that semi-dry milling as an environmental, efficient, and convenient method could provide rice flour suitable to prepare SRCs. In view of sustainable and economic development, semi-dry milling could be a good alternative method in industrial production.

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Safety Information
Symbol : GHS05, GHS08
Signal Word : Danger
Hazard Statements : H314, H315, H319, H334
Precautionary Statements : P260, P261, P264, P280, P284, P301+P330+P331, P302+P352, P303+P361+P353, P304+P340, P342+P311, P501
Safety Data Sheet
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