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Resistant Starch Assay Kit (Rapid)

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00:08   Introduction
01:09    Principle
02:35    Reagent Preparation
06:03    Weighing of samples
06:34    Hydrolysis & Solubilization of Digestible Starch
07:53    Waterbath Alignment
08:22    Ethanol washing & centrifugation
10:26    Procedure
14:26     Measurement of Digestible, Non-Resistant Starch
16:57     Calculations

Resistant Starch Assay Kit Rapid K-RAPRS Scheme
Product code: K-RAPRS

100 assays per kit

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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 mg of D-glucose per assay
Limit of Detection: 0.036 g/100 g
Reaction Time (min): ~ 5 h
Application examples: Plant materials, starch samples and other materials.
Method recognition: An updated modification of: AACC Method 32-40.01, AOAC Method 2002.02 and CODEX Method Type II

The Resistant Starch Assay Kit (Rapid) method is suitable for the analysis of resistant starch in pure starch, cereal and legume seeds and food samples.

This method is an update of the method of McCleary et al1 (AOAC Method 2002.02, AACC Method 32-40.01) employing incubation conditions similar to those used in AOAC Method 2017.16 for dietary fiber. The enzyme mixture employed [pancreatic α-amylase (PAA) and amyloglucosidase (AMG)] are those used by Englyst et al.2 except that both enzymes have been purified, standardised and stabilised. Digestion is performed using saturating levels of PAA and AMG with stirring at pH 6 and 37oC for 4 h, to simulate in vivo conditions in the human small intestine. Recent studies on the hydrolysis of “newer” resistant starch materials such as phosphate crosslinked starch (RS4) indicated that these incubation conditions are an essential requirement to obtain meaningful physiologically relevant values for RS.

The incubation conditions parallel those used in AOAC Method 2017.16, a new, rapid integrated procedure for the measurement of total dietary fiber (Megazyme method K-RINTDF). This method is physiologically based and designed to service the definition of DF announced by Codex Alimentarius in 2009.

1. McCleary, B. V., McNally, M. & Rossiter, P. (2002).  Measurement of Resistant Starch by Enzymic Digestion in Starch and Selected Plant Materials - Collaborative Study.  J. AOAC Int., 85, 1103-1111.

2. Englyst, H. N., Kingman, S. M. & Cummings, J. H. (1992).  Classification and measurement of nutritionally important starch fractions.  Eur. J. Clin. Nutr., 46 (Suppl. 2), S33-S50.

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Scheme-K-RAPRS RAPRS Megazyme

Validation of Methods
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance Validation Report
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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Rejuvenated Brewer's Spent Grain: The impact of two BSG-derived ingredients on techno-functional and nutritional characteristics of fibre-enriched pasta.

Sahin, A. W., Hardiman, K., Atzler, J., Vogelsang-O'Dwyer, M., Valdeperez, D., Münch, S., Cattaneo, G., O’Riordan, P. & Arendt, E. K. (2021). Innovative Food Science & Emerging Technologies, 68, 102633.

Brewer's Spent Grain (BSG), rich in fibre and protein is mostly used for animal feed but has great potential to be used as an ingredient for cereal based products. Originated from BSG, the two ingredients EverVita Fibra (EVF) high in fibre; and EverVita Pro (EVP) high in protein, were used to produce fibre-enriched pasta and compared to semolina, wholemeal flour and a commercial fibre-rich pasta. Analysis of gluten network development and pasting properties revealed the formation of a stronger network by the incorporation of EVP resulting in a compact pasta structure which led to a higher pasta firmness and tensile strength and a decrease in predicted glycaemic index compared to the controls. EVF resulted in an inferior product compared to EVP but was comparable to the semolina control. Hence, EVF and EVP have the potential to increase nutritional value of pasta while maintaining or even improving pasta quality and encouraging the recycling of by-streams for food production.

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Roles of Waxy and Soluble Starch Synthase Iia Alleles in Determining Different Type Resistant Starch Contents of Rice.

You, H., Zhang, O., Liang, X., Liang, C., Chen, Y., & Xiang, X. (2020). In Press.

Background: Resistant Starch (RS) is a functional starch that has functions of regulating diabetes, hypertension and obesity. The effects of most starch synthesis-related genes (SSRGs) on RS content and their relationships are largely unknown. Result: In current study, ninety-nine lines from a recombinant inbred line were selected to investigate the effects of SSRGs on the RS content in different process status. Results revealed that RS content decreased dramatically after cooking, but it did not increase significantly after cooling for 7 days. And RS was closely related to many indexes of physicochemical properties, but was not correlated with granule size. Waxy ( Wx ) played an important role in controlling RS content and Wx a could elevate RS content in raw milled rice, cooked rice and retrograded rice. Soluble starch synthase IIa ( SSIIa ) had an impact on RS2, and RS2 content of indica SSIIa were significantly higher than that of japonica SSIIa ( SSIIaj ). Moreover, interaction of Wx and SSIIa was responsible for variations of RS content in three sample types, RS2 and volume proportion of different size starch granules. Conclusions: Wx and SSIIa together significantly regulate different types content of RS in rice, but SSIIa only affects RS2. Wx a - SSIIaj is favorable to forming large-diameter starch granules.

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Leuconostoc citreum TR116 as a Microbial Cell Factory to Functionalise High-Protein Faba Bean Ingredients for Bakery Applications.

Hoehnel, A., Bez, J., Sahin, A. W., Coffey, A., Arendt, E. K. & Zannini, E. (2020). Foods, 9(11), 1706.

Grain legumes, such as faba beans, have been investigated as promising ingredients to enhance the nutritional value of wheat bread. However, a detrimental effect on technological bread quality was often reported. Furthermore, considerable amounts of antinutritional compounds present in faba beans are a subject of concern. Sourdough-like fermentation can positively affect baking performance and nutritional attributes of faba bean flours. The multifunctional lactic acid bacteria strain Leuconostoc citreum TR116 was employed to ferment two faba bean flours with different protein contents (dehulled flour (DF); high-protein flour (PR)). The strain’s fermentation profile (growth, acidification, carbohydrate metabolism and antifungal phenolic acids) was monitored in both substrates. The fermentates were applied in regular wheat bread by replacing 15% of wheat flour. Water absorption, gluten aggregation behaviour, bread quality characteristics and in vitro starch digestibility were compared to formulations containing unfermented DF and PR and to a control wheat bread. Similar microbial growth, carbohydrate consumption as well as production of lactic and acetic acid were observed in both faba bean ingredients. A less pronounced pH drop as well as a slightly higher amount of antifungal phenolic acids were measured in the PR fermentate. Fermentation caused a striking improvement of the ingredients’ baking performance. GlutoPeak measurements allowed for an association of this observation with an improved gluten aggregation. Given its higher potential to improve protein quality in cereal products, the PR fermentate seemed generally more promising as functional ingredient due to its positive impact on bread quality and only moderately increased starch digestibility in bread.

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Physico-chemical properties of flour, dough and bread from wheat and hydrothermally-treated finger millet.

Onyango, C., Luvitaa, S. K., Unbehend, G. & Haase, N. (2020). Journal of Cereal Science, 102954.

Hydrothermally-treated (HTT) finger millet was prepared by tempering the grains twice with water (10:1) followed by incubation at about 25-30°C in a woven polypropylene sack for 10 days. Hydrothermally-treated finger millet was darker and had higher α-amylase activity and lower starch digestibility than native (NAT) grains. The HTT finger millet was composited with wheat flour and used to prepare bread. Composite dough had higher dough stability, dough development time and degree of softening but lower dough energy, extensibility and resistance to extension than WHE dough. The higher specific volume and lower crumb firmness and chewiness of WHE-HTT compared to WHE-NAT bread was attributed to the high α-amylase activity and water absorption capacity of HTT finger millet. Wheat-HTT bread had higher dietary fibre, phytate and phenolic acid content but the same starch and protein digestibility as WHE bread.

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Design and synthesis of modified and resistant starch-based oil-in-water emulsions.

Jain, S., Winuprasith, T. & Suphantharika, M. (2019). Food Hydrocolloids, 89, 153-162.

In this study, rice starch was modified by different modifications that included debranching (DB), esterification with octenyl succinic anhydride (OSA), debranching followed by OSA modification (DBOS) and OSA modification followed by debranching (OSDB). Following modification, the resistant starch content and emulsifying properties of the modified starches markedly increased in comparison with the native starch with the DBOS starch having the highest resistant starch content and the best emulsifying properties. This can be attributed to its higher degree of OSA substitution resulting in an enhanced amphiphilic character and better emulsion stability. Microstructure, physical stability and rheological properties of oil-in-water emulsions using these modified rice starches were also investigated and it was observed that emulsions stabilized by OSA and DBOS starches exhibited smaller oil droplets, were more stable to creaming and possessed stronger three-dimensional structure when compared with the other modified starch-based emulsions. Since the DBOS starch was found to demonstrate the highest resistant starch content and better emulsifying properties than the other starches, it was then chosen for optimization of its concentration to give the best emulsion characteristics and stability. The emulsions stabilized by the DBOS starch at a concentration of 4% or 5% (w/w) were found to be more stable and very little phase separation was observed after a storage period of 30 days.

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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, P501
Safety Data Sheet
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