A simple microplate-based method for the determination of α-amylase activity using the glucose assay kit (GOD method). (15th November 2016)

• Record Type: Journal Article
• Authors: Visvanathan, Rizliya; Jayathilake, Chathuni; Liyanage, Ruvini
• Is Part Of: Food chemistry. Volume 211(2016)
• Page Start: 853
• Page End: 859
• Publication Date: 2016-11-15
• Subjects: Gallic acid (PubChem CID: 370) -- (+)-Catechin hydrate (PubChem CID: 9064) -- Rutin hydrate (PubChem CID: 45479757) -- Maltose monohydrate (PubChem CID: 16211336) -- Anhydrous dextrose (PubChem CID: 5793) -- Sodium phosphate dibasic (PubChem CID: 24203) -- Sodium phosphate monobasic (PubChem CID: 23672064) -- Starch soluble (PubChem CID: 439341) -- 3, 5-Dinitrosalicylic acid (PubChem CID: 11873) -- Potassium sodium tartrate tetrahydrate (PubChem CID: 2724148); Glucose oxidase -- α-Amylase -- Maltose -- Amylase inhibitors -- DNSA; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2016.05.090
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: A new method was established for α-amylase activity determination. The method is simple and less laborious. Total assay time is 40 min including the pre-incubation steps. The developed method is accurate, quantitative, precise and highly reproducible. The high-throughput technique is also suitable for kinetic studies. Abstract: For the first time, a reliable, simple, rapid and high-throughput analytical method for the detection and quantification of α-amylase inhibitory activity using the glucose assay kit was developed. The new method facilitates rapid screening of a large number of samples, reduces labor, time and reagents and is also suitable for kinetic studies. This method is based on the reaction of maltose with glucose oxidase (GOD) and the development of a red quinone. The test is done in microtitre plates with a total volume of 260 μL and an assay time of 40 min including the pre-incubation steps. The new method is tested for linearity, sensitivity, precision, reproducibility and applicability. The new method is also compared with the most commonly used 3, 5-dinitrosalicylic acid (DNSA) method for determining α-amylase activity.