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Total Starch Assay Kit (AA/AMG)

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0:05  Introduction
2:11    Principle
3:26   Reagent and Sample Preparation
6:22   RTS Method (Solubilization & Hydrolysis of Starch)
12:56  RTS-NaOH Method (Solubilization & Hydrolysis of Starch)
15:41   Calculation

Total Starch Assay Kit AA/AMG K-TSTA TSTA
   
Product code: K-TSTA-100A

Content:

€249.00

100 assays

Prices exclude VAT

Available for shipping

Content: 50 assays / 100 assays
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: Total Starch
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 510
Signal Response: Increase
Linear Range: 4 to 100 μg of D-glucose per assay
Limit of Detection: 0.18 g/100 g total starch “as is”
Total Assay Time: ~ 90 min
Application examples: Cereal flours, food products and other materials.
Method recognition: AACC Method 76-13.01, AOAC Method 996.11, ICC Standard Method No. 168 and RACI Standard Method

The Total Starch (AA/AMG) test kit is used for the measurement and analysis of total starch in cereal flours and food products.  This kit now contains an improved α-amylase that allows the amylase incubations to be performed at pH 5.0 (as well as pH 7.0). 

See our full range of dietary fiber and starch assay kits.


Total Starch Assay Kit AA AMG K-TSTA Scheme

Advantages
  • Very competitive price (cost per test) 
  • All reagents stable for > 2 years after preparation 
  • Rapid reaction 
  • 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 Booklet 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

Measurement of Starch: Critical evaluation of current methodology.

McCleary, B. V., Charmier, L. M. J. & McKie, V. A. (2018). Starch‐Stärke, 71(1-2), 1800146.

Most commonly used methods for the measurement of starch in food, feeds and ingredients employ the combined action of α‐amylase and amyloglucosidase to hydrolyse the starch to glucose, followed by glucose determination with a glucose oxidase/peroxidase reagent. Recently, a number of questions have been raised concerning possible complications in starch analytical methods. In this paper, each of these concerns, including starch hydrolysis, isomerisation of maltose to maltulose, effective hydrolysis of maltodextrins by amyloglucosidase, enzyme purity and hydrolysis of sucrose and β‐glucans have been studied in detailed. Results obtained for a range of starch containing samples using AOAC Methods 996.11 and 2014 .10 are compared and a new simpler format for starch measurement is introduced. With this method that employs a thermostable α-amylase (as distinct from a heat stable α-amylase) which is both stable and active at 100°C and pH 5.0, 10 samples can be analysed within 2 h, as compared to the 6 h required with AOAC Method 2014.10.

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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 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 a simplified assay for total starch in cereal products (AACC Method 76-13).

McCleary, B. V., Gibson, T. S. & Mugford, D. C. (1997). Cereal Foods World, 42, 476-480.

A procedure for the quantitative analysis of total starch in plant materials has been developed and subjected to a comprehensive interlaboratory study involving 32 laboratories, in accordance with the protocol for collaborative studies recommended by American Association of Cereal Chemists and AOAC International. The method involved treatment of a sample at approximately 95°C with thermostable α-amylase to obtain starch depolymerization and solubilisation. The slurry is then treated with purified amyloglucosidase to give quantitative hydrolysis of the starch fragments to glucose, which is measured with glucose oxidase/peroxidase reagent. Test samples used in the interlaboratory study included modified and native starches, cereal flours and brans, processed cereal products, animal feeds, and plant material. Results were statistically analysed according to AOAC International guidelines (1). The procedure was shown to be highly repeatable (relative standard deviation 2.1-3.9%) and reproducible (relative standard deviation 2.9-5.0%), and on the basis of these results has gained first approval status with AACC (AACC Method 76-13) and approval as AOAC Method 986.11. The method is more robust than a method previously reported (AACC Method 76-12), and 20 samples can be analysed within 2 hr.

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Megazyme publication
Total starch measurement in cereal products: interlaboratory evaluation of a rapid enzymic test procedure.

McCleary, B. V., Gibson, T. S., Solah, V. & Mugford, D. C. (1994). Cereal Chemistry, 71(5), 501-505.

The precision of an enzymatic procedure for analysis of total starch in cereal flours and products was determined in a comprehensive inter-laboratory study involving 29 laboratories. Test samples represented a range of sample types, including modified and native starches, cereal flours and brans, processed cereal products, animal feeds, and plant material. Results were statistically analyzed according to AOAC guidelines. The procedure was shown to be highly repeatable (relative standard deviation 1.5-7.3%) and reproducible (relative standard deviation 4.1-11.3%). It is now available, in a slightly modified form, as an assay kit. The assay, therefore, provides a convenient alternative to existing procedures for quantitative measurement of starch in cereal products.

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Megazyme publication
Quantitative measurement of total starch in cereal flours and products.

McCleary, B. V., Solah, V. & Gibson, T. S. (1994). Journal of Cereal Science, 20(1), 51-58.

A rapid and quantitative method has been developed for the determination of total starch in a wide range of materials, including high-amylose maize starches and food materials containing resistant starch. The method allows the analysis of 20 samples in 3 h. A single assay can be performed in 2 h. For a range of samples, the total starch values obtained with this method were significantly higher than those obtained with current standard methods. Two assay formats have been developed. In assay format 1, the sample is incubated solubilised with the chaotropic agent dimethyl sulphoxide (DMSO) to gelatinise the starch, which is then solubilised and partially depolymerised by controlled incubation at ∼ 100°C with a defined level of thermostable alpha-amylase. This allowed near-complete solubilisation of most starches. The remaining starch is then solubilised and the starch fragments are converted to maltose and maltotriose by the combined action of highly purified pullulanase and beta-amylase. After volume adjustment and filtration (if necessary), the maltooligo-saccharides are hydrolysed by high-purity amyloglucosidase to glucose, which is measured with a glucose oxidase/peroxidase reagent. This assay format gave quantitative starch determination in all native starch samples, including high-amylose maize starches. In assay format 2, which is applicable to most starches and cereal flours, the DMSO pre-treatment step is omitted. Samples containing glucose and/or maltosaccharide are pre-washed with aqueous ethanol before analysis.

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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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