title: An amperometric bienzymatic cholesterol biosensor based on functionalized graphene modified electrode and its electrocatalytic activity towards total cholesterol determination.
abstract: Cholesterol oxidase (ChOx) and cholesterol esterase (ChEt) have been covalently immobilized onto functionalized graphene (FG) modified graphite electrode. Enzymes modified electrodes were characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). FG accelerates the electron transfer from electrode surface to the immobilized ChOx, achieving the direct electrochemistry of ChOx. A well defined redox peak was observed, corresponding to the direct electron transfer of the FAD/FADH(2) of ChOx. The electron transfer coefficient (α) and electron transfer rate constant (K(s)) were calculated and their values are found to be 0.31 and 0.78 s(-1), respectively. For the free cholesterol determination, ChOx-FG/Gr electrode exhibits a sensitive response from 50 to 350 μM (R=-0.9972) with a detection limit of 5 μM. For total cholesterol determination, co-immobilization of ChEt and ChOx on modified electrode, i.e. (ChEt/ChOx)-FG/Gr electrode showed linear range from 50 to 300 μM (R=-0.9982) with a detection limit of 15 μM. Some common interferents like glucose, ascorbic acid and uric acid did not cause any interference, due to the use of a low operating potential. The FG/Gr electrode exhibits good electrocatalytic activity towards hydrogen peroxide (H(2)O(2)). A wide linear response to H(2)O(2) ranging from 0.5 to 7 mM (R=-0.9967) with a sensitivity of 443.25 μA mM(-1) cm(-2) has been obtained.
authors.affiliation: Chemistry Research Centre, S. S. M. R. V. Degree College, Jayanagar, Bangalore-560041, India.
authors.forename: Revanasiddappa
authors.initials: R
authors.lastname: Manjunatha
authors.forename: Gurukar
authors.initials: G
authors.lastname: Shivappa Suresh
authors.forename: Jose Savio
authors.initials: JS
authors.lastname: Melo
authors.forename: Stanislaus F
authors.initials: SF
authors.lastname: D'Souza
authors.forename: Thimmappa Venkatarangaiah
authors.initials: TV
authors.lastname: Venkatesha
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002384:Catalysis; D002784:Cholesterol / Q000032:analysis* / Q000097:blood; D002789:Cholesterol Oxidase / Q000737:chemistry / Q000378:metabolism*; D058266:Dielectric Spectroscopy; D004566:Electrodes; D004579:Electron Transport; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D002787:Sterol Esterase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D006108:Graphite; D002784:Cholesterol; D006861:Hydrogen Peroxide; D002789:Cholesterol Oxidase; D002787:Sterol Esterase