Resistant Starch Assay Kit

Reference code: K-RSTAR
SKU: 700004336

100 assays per kit

Content: 100 assays per kit
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 2 years under recommended storage conditions
Analyte: Resistant Starch
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 510
Signal Response: Increase
Linear Range: 4 to 100 μg of glucose per assay
Limit of Detection: 0.036 g/100 g
Reaction Time (min): ~ 17 h
Application examples: Plant materials, starch samples and other materials.
Method recognition: AACC Method 32-40.01, AOAC Method 2002.02 and CODEX Method Type II

The Resistant Starch Assay Kit for the measurement and analysis of resistant starch in plant materials and starch samples. Official analysis methods: AOAC Method 2002.02, AACC Method 32-40.01, CODEX Type II Method.

By definition, resistant starch (RS) is that portion of the starch that is not broken down by human enzymes in the small intestine. It enters the large intestine where it is partially or wholly fermented. RS is generally considered to be one of the components that make up total dietary fiber (TDF).

See our full range of starch and dietary fiber products.

Scheme-K-RSTAR RSTAR Megazyme

Advantages
  • Very cost effective 
  • All reagents stable for > 2 years after preparation 
  • Only enzymatic kit available 
  • Measures enzyme resistant starch 
  • Simple format 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Standard included
Validation of Methods
Documents
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance
Publications
Megazyme publication

Measurement of available carbohydrates, digestible, and resistant starch in food ingredients and products.

McCleary, B. V., McLoughlin, C., Charmier, L. M. J. & McGeough, P. (2019). Cereal Chemistry, 97(1), 114-137.

Background and objectives: The importance of selectively measuring available and unavailable carbohydrates in the human diet has been recognized for over 100 years. The levels of available carbohydrates in diets can be directly linked to major diseases of the Western world, namely Type II diabetes and obesity. Methodology for measurement of total carbohydrates by difference was introduced in the 1880s, and this forms the basis of carbohydrate determination in the United States. In the United Kingdom, a method to directly measure available carbohydrates was introduced in the 1920s to assist diabetic patients with food selection. The aim of the current work was to develop simple, specific, and reliable methods for available carbohydrates and digestible starch (and resistant starch). The major component of available carbohydrates in most foods is digestible starch. Findings: Simple methods for the measurement of rapidly digested starch, slowly digested starch, total digestible starch, resistant starch, and available carbohydrates have been developed, and the digestibility of phosphate cross‐linked starch has been studied in detail. The resistant starch procedure developed is an update of current procedures and incorporates incubation conditions with pancreatic α‐amylase (PAA) and amyloglucosidase (AMG) that parallel those used AOAC Method 2017.16 for total dietary fiber. Available carbohydrates are measured as glucose, fructose, and galactose, following complete and selective hydrolysis of digestible starch, maltodextrins, maltose, sucrose, and lactose to glucose, fructose, and galactose. Sucrose is hydrolyzed with a specific sucrase enzyme that has no action on fructo‐oligosaccharides (FOS). Conclusions: The currently described “available carbohydrates” method together with the total dietary fiber method (AOAC Method 2017.16) allows the measurement of all carbohydrates in food products, including digestible starch. Significance and novelty: This paper describes a simple and specific method for measurement of available carbohydrates in cereal, food, and feed products. This is the first method that provides the correct measurement of digestible starch and sucrose in the presence of FOS. Such methodology is essential for accurate labeling of food products, allowing consumers to make informed decisions in food selection.

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Megazyme publication

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.

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.

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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 resistant starch.

McCleary, B. V. & Monaghan, D. A. (2002). Journal of AOAC International, 85(3), 665-675.

A robust and reliable method was developed to measure resistant starch (RS), i.e., starch that enters the large intestine. In vivo conditions were reflected as much as possible while a user-friendly format was maintained. Parameters investigated included α-amylase concentration, pH of incubation, maltose inhibition of α-amylase, the need for amyloglucosidase inclusion, the effect of shaking and stirring on determined values, and problems in recovering and analyzing the RS-containing pellet. The RS values obtained were in good agreement with published in vivo data. An interlaboratory evaluation of the method has been completed (First Action Method 2002.02).

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Megazyme publication

Measurement of resistant starch by enzymatic digestion in starch and selected plant materials: Collaborative study.

McCleary, B. V., McNally, M. & Rossiter, P. (2002). Journal of AOAC International, 85(5), 1103-1111.

Interlaboratory performance statistics was determined for a method developed to measure the resistant starch (RS) content of selected plant food products and a range of commercial starch samples. Food materials examined contained RS (cooked kidney beans, green banana, and corn flakes) and commercial starches, most of which naturally contain, or were processed to yield, elevated RS levels. The method evaluated was optimized to yield RS values in agreement with those reported for in vivo studies. Thirty-seven laboratories tested 8 pairs of blind duplicate starch or plant material samples with RS values between 0.6 (regular maize starch) and 64% (fresh weight basis). For matrixes excluding regular maize starch, repeatability relative standard deviation (RSDr) values ranged from 1.97 to 4.2%, and reproducibility relative standard deviation (RSDR) values ranged from 4.58 to 10.9%. The range of applicability of the test is 2-64% RS. The method is not suitable for products with <1% RS (e.g., regular maize starch; 0.6% RS). For such products, RSDr and RSDR values are unacceptably high.

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Megazyme publication

Two issues in dietary fiber measurement.

McCleary, B. V. (2001). Cereal Foods World, 46, 164-165.

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.

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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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Publication

Structural, physicochemical properties and noodle-making potential of quinoa starch and type 3, type 4, and type 5 quinoa resistant starch. 

Niu, H., Zhao, F., Ji, W., Ma, L., Lu, B., Yuan, Y. & Yue, T. (2024). International Journal of Biological Macromolecules, 258, 128772.

This study prepared type 3, type 4, and type 5 quinoa resistant starch (QRS3, QRS4, and QRS5) from quinoa starch (QS), compared their structural and physicochemical properties and evaluated their noodle-making potential. The results showed that the molecular weight of QRS3 decreased, the number of short-chain molecules increased, and its crystal type changed to B-type after gelatinization, enzymatic hydrolysis, and retrogradation. QRS4 is a phosphorylated cross-linked starch, with a surface morphology, particle size range, and crystal type similar to QS, but displaying modified thermodynamic properties. QRS5 is a complex of amylose and palmitic acid. It displays typical V-type crystals, mainly composed of long chain molecules and primarily exhibits a block morphology. The noodles prepared by replacing 20 % wheat flour with QS, QRS3 and QRS5 have higher hardness and are suitable for people who like elasticity and chewiness. QRS4 noodles are softer and suitable for people like elderly and infants who prefer soft foods. In conclusion, significant differences were evident between the fine structures, crystal types, physicochemical properties and potential applications of QS and the three QRSs. The results may expand the application of QS and QRS in the food and pharmaceutical industries.

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Publication

Native And Modified Sago (Metroxylon sagu) Starches as an Ingredient in The Formulation of Low Glycaemic Food Product. 

Zailani, M. A., Kamilah, H., Husaini, A. A. S. A., Seruji, A. Z. R. A. & Sarbini, S. R. (2023). Malaysian Applied Biology, 52(5), 129-136.

Native sago starch has a high content of resistant starch (RS) which is associated with low glycaemic and beneficial to individuals with obesity and diabetes. Additionally, the RS is linked to the prebiotic properties exhibited by starch. This study aimed to evaluate the predicted glycaemic index (pGI) and probiotic growth rates of food formulated with native or modified starches in the formulation of a breakfast drink. The sago starch was modified via microwave heat treatment (MHT) with different treatment duration or via pre-treatment followed by MHT. The formulation of food was performed by replacing a portion of wheat starch at percentages of 25, 50, or 75%. The pGI was determined by measuring the amount of glucose produced during in vitro digestion. Meanwhile, the probiotic growth rates were conducted by monitoring the optical density of Lactobacillus casei and Bifidobacterium lactis for 24 hr. Comparatively, food formulated with 50 and 75% starch showed lower pGI than other formulations. This was correlated with the increase of RS in food products. Meanwhile, the probiotic growth rates increase for a few of the formulations mostly with a higher pGI or low RS content which is contributed by the accessibility for fermentation to occur. In conclusion, the findings suggest the substitution of 50% wheat flour with native or modified sago starches is sufficient to increase RS content and lower the pGI of formulated food. In the future, investigation of RS components contributing to probiotic growth is needed to enable the exploration of new prebiotics with low glycaemic.

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Publication

Sustainable Strategies to Increase the Content of Protein, Unsaturated Fatty Acids and Vitamins in Tenebrio molitor Larvae Flours through Vegetable Waste Supplementation. 

López-Gámez, G., del Pino-García, R., López-Bascón, M. A., Lara-Cambil, A., Vigil-Chacón, A. & Quiles-Morales, J. L. (2023, October). Biology and Life Sciences Forum, 26(1), 58.

Tenebrio molitor larvae were fed with wheat bran and supplemented (1:1) with tomato or cucumber agricultural waste for 6 weeks. After supplementation, larvae were dried in a pilot-infrared oven (68°C for 4 h) and ground to obtain the flours. The quality attributes and nutritional value of insect flours differed based on the supplemented diet. Unsaturated fatty acids, proteins, starches and certain vitamins were enhanced in flours from supplemented larvae. Therefore, tomato and cucumber waste can be revalorised as supplements of T. molitors’ conventional diet to obtain insect flours with higher nutritional values and acceptable-quality attributes.

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Publication

Effect evaluation of banana on improving hyperglycemia and hypertension in diabetic spontaneously hypertensive rats.

Lin, H. L., Tseng, Y. H., Yeh, C. C., Chen, Y. M. & Shen, K. P. (2023). Natural Product Communications, 18(11), 1934578X231213225.

Background: This study aimed to evaluate the protective effects of dried and milled banana (BN) against streptozocin (STZ), nicotinamide (NA), and high-fat diet (HFD)-induced diabetes mellitus (DM) in 8-week-old spontaneously hypertensive rats (SHRs). Methods: First, we analyzed the resistance starch contents, anti-oxidant compositions and activity, and α-glucosidase inhibitory effects of the BN. Then the DM animal model was performed. The SHRs were randomly divided into 3 groups. The control group was fed normal chow, the DM group was injected with STZ/NA and fed HFD, and the DM/BN group was injected with STZ/NA and fed HFD having the carbohydrate content replaced with BN. Results: After 4 weeks, hyperglycemia and hypoinsulinemia occurred; moreover, HbA1c levels were higher and glucagon-like peptide 1 concentrations were lower, but blood pressure and heart rate did not worsen in the DM group. Related glucose metabolism and vasorelaxation proteins were disrupted by DM. However, in the DM/BN group, all of the above biochemical parameters were ameliorated by improving glucose homeostasis and clearly brought about improvements in blood pressure and heart rate as well. Conclusion: Together, the functional compositions of banana may help to improve DM, hypertension, and related complications.

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Publication

Investigation of freezing temperature and time to improve resistant starch content and quality of pure mung bean starch vermicelli.

Nguyen, T. P., Rumpagaporn, P. & Songsermpong, S. (2023). Australian Journal of Crop Science, 17(9), 693-698.

Mung bean vermicelli is one of the commonly consumed styles of noodles because of its convenience in Asian countries as well as being a good source of resistant starch (RS). It was hypothesized that suitable freezing conditions could produce mung bean vermicelli with a greater RS content and good cooking quality compared to the commercially available product. Therefore, this study investigated the optimum freezing conditions to increase the RS content in mung bean vermicelli. After extrusion, boiling, and cooling, the drained vermicelli was incubated at 4 C for 1 h 30 min before using different freezing conditions by varying the processing temperature (-5, -10, -15, or -20 &#61551;C) and time (6, 12, 18, or 20 h for each temperature).The highest RS content was observed in dried mung bean vermicelli (12.83% at -10 &#61551;C for 18 h; V1018), whereas the commercial vermicelli had the lowest RS content (9.73%). The cooking time, water absorption, volume increase, and resistant starch content of cooked V1018 were higher with values of 12.69 min, 502.97%, 19.13% and 7.32%, respectively, than those of the commercial vermicelli (8.56 min, 347.34%, 15.19% and 5.58%, respectively). Furthermore, the cooking loss, tensile strength, and elasticity of V1018 were 1.08%, 0.0101 N, and 11.96 mm, respectively, which were lower than for the commercial vermicelli (1.96%, 1.16-folds, and 2.16-folds, respectively). The color of dried V1018 vermicelli had more lightness, redness, and yellowness, while the cooked V1018 vermicelli had more lightness, greenness, and blueness than the respective commercial products. Hence, incubating at 4 C for 1 h 30 min, followed by freezing at -10 C for 18 h was recommended to produce pure mung bean starch vermicelli with a high resistant starch content and good quality.

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Publication

Barley SS2a single base mutation at the splicing site led to obvious change in starch.

Bang, Wang., Jing, Liu., Chen, X. L., Qiang, XU., ZHANG, Y. Z., Dong, H. X., Tang, H. P., Qi, P. F., Deng, M., Ma, J., Wnag, J. R., Chen, G. Y., Wei, Y. M., Zheng, Y. L. & Jiang, Q. T. (2023). Journal of Integrative Agriculture, In Press.

Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes. Resistant starch is not digested in the small intestine, thus preventing the rapid rise of the glycemic index. Starch synthase 2a (SS2a), a key enzyme in amylopectin biosynthesis, has significant effects on starch structure and properties. In this study, we identified an ss2a null mutant (M3-1413) with a single base mutation from an ethyl methane sulfonate (EMS)-mutagenized population of barley. The mutation was located at the 3´ end of the first intron of the RNA splicing receptor (AG) site, resulting in abnormal RNA splicing and two abnormal transcripts of ss2a, which caused the inactivation of the SS2a gene. The starch structure and properties were significantly altered in the mutant, with M3-1413 containing decreased total starch and increased amylose and resistant starch levels. This study sheds light the effect of barley ss2a null mutations on starch properties and helps to guide new applications of barley starch to develop nutritious food products.

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Publication

Decorticated lentil malt flour: production process and use.

Cimini, A., Poliziani, A. & Moresi, M. (2023). Chemical Engineering Transactions, 102, 121-126.

In this work, the malting process of lentil seeds (Lens culinaris) was set-up to minimize their anti-nutrient content. The first (water steeping) and second (germination) process steps were studied in a 1.2-kg bench-top plant at 25°C. After 2-h steeping about 98.8% of seeds sprouted. As the germination process was prolonged for 72 h, the flatulence-inducing raffinose or phytic acid content was reduced by 94% or 63%, respectively. The third process step (kilning), carried out under fluent dry air at 50°C for 48 h and at 75°C for 3 h, gave rise to a gold metallic yellow-lentil malt, the cotyledons of which were cyclonically recovered and finally milled. The resulting decorticated yellow-lentil malt flour was used to prepare a fresh egg pasta high in raw protein (28±2 g/100 g), low in phytate (0.46±0.03 g/100 g) and in vitro glycemic index (38%), and approximately zero oligosaccharides.

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Publication

Compositional characterization of starch, proteins and lipids of long bean, dwarf long bean, mung bean and French bean seed flours.

Azmah, U. N., Makeri, M. U., Bagirei, S. Y. & Shehu, A. B. (2023). Measurement: Food, 12, 100111.

Four underutilized legumes comprised of mung bean, long bean, dwarf long bean and French bean were investigated for their total starch, dietary fibre, amylose, amylopectin and resistant contents along with proteins and lipids profiles employing standard methods in order to explore appropriate end uses. Results showed significant variations (p<0.05) in the fat, protein, moisture, ash, total dietary fibre, carbohydrate, starch, amylose and amylopectin contents of all the four legume seeds Flours. Mung bean, long bean, dwarf long bean, and French bean showed carbohydrates contents of 55.24, 49.63, 48.12 and 45.56 g/100 g, respectively. Dwarf long bean, long bean and mung bean recorded total starch of 38.39, 49.78 and 57.68 g/100 g, respectively. Whereas French bean had the highest total dietary fibre of 16.91 g/100 g and dwarf long bean the highest amylopectin (86.67 g/100 g), both dwarf long bean and long bean recorded insoluble dietary fibre and soluble dietary fibre in the range of 7.25 to 14.58 g/100 g and 0.05 to 1.25 g/100 g, respectively, however, all the seed flours showed higher content of insoluble dietary fibre than soluble dietary fibre. Significant differences (p<0.05) observed in the total starch, resistant starch and non-resistant starch content of all the 4 samples. Non-resistant starch was the predominant constituents of the total of all the seed flours with mung bean showing the highest value (54.16 g/100 g) followed by long bean, dwarf long bean and French bean with 48.74, 37.73 and 16.88 g/100 g, respectively. Though the beans had low amylose contents, their amino acids score and patterns surpassed the FAO reference proteins and their saturated and unsaturated fatty acids ratio falls within FAO allowed limits. This study highlights the hidden potentials of these non-conventional legumes as sources of quality proteins, functional foods, healthy fats and dietary fibres.

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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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