5:15 PM - 5:30 PM
[2Dp-10] Development of a Novel Colorimetric Fructose Assay Kit Using Enzymatic Method
Keywords:fructose, assay kit, enzymatic method, colorimetric determination
[Purpose] Currently used fructose assay kits need calculations and heating procedure, making it difficult for non-experts to use in food handling environments. Therefore, the aim of this study was to develop a novel and convenient fructose assay kit that does not require heating and complicated operations.
[Methods] A two-step reaction was designed. Firstly, the sample was treated with Solution A to remove glucose and ascorbic acid, which inhibits color reactions. Secondly, by glucose isomerase in Solution B, fructose was converted to glucose, which was immediately converted to pigment using glucose oxidase, peroxidase, coupling compounds and Trinder’s reagents. We explored the conditions required for all these reactions to be proceeded at room temperature and constructed a calibration curve by rate assay. A simple assay kit consisting of Solution A, Solution B and standard solution (fructose) was conducted. The substrate specificity for monosaccharides was assessed. Fructose concentrations of actual food samples were measured using the new kit and conventional methods to compare the results.
[Results] A kit capable of measuring fructose concentration at room temperature was successfully developed. A good linear relationship between absorbance and fructose concentration was obtained within the range of 50-500 mg/L. The kit showed low activity against mannose and galactose. A high correlation was observed between the fructose concentrations measured by the established and newly developed methods, confirming the reliability of the new fructose assay kit.
[Methods] A two-step reaction was designed. Firstly, the sample was treated with Solution A to remove glucose and ascorbic acid, which inhibits color reactions. Secondly, by glucose isomerase in Solution B, fructose was converted to glucose, which was immediately converted to pigment using glucose oxidase, peroxidase, coupling compounds and Trinder’s reagents. We explored the conditions required for all these reactions to be proceeded at room temperature and constructed a calibration curve by rate assay. A simple assay kit consisting of Solution A, Solution B and standard solution (fructose) was conducted. The substrate specificity for monosaccharides was assessed. Fructose concentrations of actual food samples were measured using the new kit and conventional methods to compare the results.
[Results] A kit capable of measuring fructose concentration at room temperature was successfully developed. A good linear relationship between absorbance and fructose concentration was obtained within the range of 50-500 mg/L. The kit showed low activity against mannose and galactose. A high correlation was observed between the fructose concentrations measured by the established and newly developed methods, confirming the reliability of the new fructose assay kit.