title: Colorimetric Glucose Assay Based on Magnetic Particles Having Pseudo-peroxidase Activity and Immobilized Glucose Oxidase.
abstract: Magnetic particles (MPs) are currently used as a suitable alternative for peroxidase in the construction of novel biosensors, analytic and diagnostic methods. Their better chemical and thermal stabilities predestine them as appropriate pseudo-enzymatic catalysts. In this point of view, our research was focused on preparation of simply and fast method for immobilization of glucose oxidase onto surface of MPs with peroxidase-like activity. Spectrophotometric method (wavelength 450 nm) optimized for glucose determination using modified MPs has been successfully developed. Concentration curve for optimization of method was assayed, and Michaelis-Menten constant (K m) calculated, maximum reaction rate (V max), limit of detection, and correlation coefficient were determined to be 0.13 mmol/l (2.34 mg/dl), 1.79 pkat, 3.74 µmol/l (0.067 mg/dl), and 0.996, respectively. Interferences of other sugars such as sucrose, sorbitol, deoxyribose, maltose, and fructose were determined as well as effect of substances presenting in plasma (ascorbic acid, reduced glutathione, trolox, and urea). Results in comparison with positive and negative controls showed no interferences of the other sugars and no influence of plasma substances to measuring of glucose. The constructed method showed corresponding results with linear dependence and a correlation coefficient of 0.997. Possibility of repeated use of modified MPs was successfully proved.
authors.affiliation: Faculty of Military Health Sciences, University of Defence, Trebesska 1575, Hradec Kralove, 500 01, Czech Republic.
authors.forename: Pavla
authors.initials: P
authors.lastname: Martinkova
authors.affiliation: Department of Chemical Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.
authors.forename: Radka
authors.initials: R
authors.lastname: Opatrilova
authors.affiliation: Laboratory of Structural Biology and Proteomics, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Palackeho tr 1/1946, 613 00, Brno, Czech Republic. kruzliakpeter@gmail.com.
authors.forename: Peter
authors.initials: P
authors.lastname: Kruzliak
authors.affiliation: Institute of Geotechnics, Slovak Academy of Sciences, Kosice, Slovak Republic.
authors.forename: Igor
authors.initials: I
authors.lastname: Styriak
authors.affiliation: Faculty of Military Health Sciences, University of Defence, Trebesska 1575, Hradec Kralove, 500 01, Czech Republic. miroslav.pohanka@gmail.com.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Pohanka
mesh_terms: D001786:Blood Glucose / Q000378:metabolism*; D003124:Colorimetry / Q000379:methods*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005260:Female; D005949:Glucose Oxidase / Q000378:metabolism*; D006801:Humans; D008280:Magnetics; D008297:Male; D010544:Peroxidases / Q000378:metabolism*; D013053:Spectrophotometry
chemical_list: D001786:Blood Glucose; D004800:Enzymes, Immobilized; D005949:Glucose Oxidase; D010544:Peroxidases
keywords: Glucose oxidase; Glucose sensor; Magnetic particles; Mimic peroxidase; Spectrophotometry