title: A novel horseradish peroxidase biosensor towards the detection of dopamine: a voltammetric study.
abstract: A polymerized film of glycine (Gly) was prepared on the surface of carbon paste electrode (CPE) through the cyclic voltammetry (CV) technique. A novel biosensor for the determination of dopamine (DA) has been constructed based on horseradish peroxidase (HRP) and multiwalled carbon nanotubes (MWCNTs) immobilizing on Poly (Gly)/CPE through silica sol-gel (SiSG) entrapment. CV measurements were employed in order to understand the feasibility of poly (Gly) as an electron carrier between the immobilized peroxidase and the surface of CPE. By using differential pulse voltammetry (DPV) the calibration curves of DA was obtained in the range of 15-865 μM. The limit of detection (LOD) and limit of quantification (LOQ) of DA was found to be 6×10⁻⁷ M and 2×10⁻⁶ M respectively. The apparent Michaelis-Menten constant (Km(app)) was found to be 0.5 mM and illustrated that the good biological activity of the fixed enzyme. Electrochemical impedance spectroscopy (EIS) results confirmed the rapid electron transfer and also the immobilization of enzyme on the electrode surface. The biosensor showed high sensitivity, selectivity and reproducibility. This method has been used to determine DA in the presence of various interferences and in clinical preparations.
authors.affiliation: Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India.
authors.forename: P
authors.initials: P
authors.lastname: Raghu
authors.affiliation: Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India. Electronic address: tmsreddysvu@gmail.com.
authors.forename: T Madhusudana
authors.initials: TM
authors.lastname: Reddy
authors.affiliation: Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India.
authors.forename: P
authors.initials: P
authors.lastname: Gopal
authors.affiliation: Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India.
authors.forename: K
authors.initials: K
authors.lastname: Reddaiah
authors.affiliation: Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India.
authors.forename: N Y
authors.initials: NY
authors.lastname: Sreedhar
mesh_terms: D015374:Biosensing Techniques / Q000379:methods* / Q000706:statistics & numerical data; D058266:Dielectric Spectroscopy; D004298:Dopamine / Q000032:analysis*; D055664:Electrochemical Techniques; D004800:Enzymes, Immobilized; D006735:Horseradish Peroxidase; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D037742:Nanotubes, Carbon; D010455:Peptides; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004800:Enzymes, Immobilized; D037742:Nanotubes, Carbon; D010455:Peptides; C011080:polyglycine; D006735:Horseradish Peroxidase; D004298:Dopamine
keywords: Carbon paste electrode; Dopamine; Glycine; Horseradish peroxidase; Multiwalled carbon nanotubes; Silica sol–gel