Covalent attachment of laccase to carboxymethyl-botryosphaeran in aqueous solution for the construction of a voltammetric biosensor to quantify quercetin
| Autor: | Adriana Gomes, Robert F.H. Dekker, Gabriel Junquetti Mattos, Elen Romão Sartori, Bruna Coldibeli, Aneli M. Barbosa Dekker |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Biosensor device
Supporting electrolyte Biophysics 02 engineering and technology Biosensing Techniques 01 natural sciences Limit of Detection Electrochemistry Physical and Theoretical Chemistry Voltammetry Glucans Detection limit Laccase Aqueous solution Chemistry 010401 analytical chemistry Water General Medicine Electrochemical Techniques 021001 nanoscience & nanotechnology 0104 chemical sciences Dielectric spectroscopy Solutions Quercetin 0210 nano-technology Biosensor Nuclear chemistry |
| Zdroj: | Bioelectrochemistry (Amsterdam, Netherlands). 135 |
| ISSN: | 1878-562X |
| Popis: | Laccase from Botryosphaeria rhodina MAMB-05 was covalently immobilized on carboxymethyl-botryosphaeran by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide (EDC/NHS) in aqueous solution. This approach was employed to fabricate a novel laccase-based biosensor to electrochemically quantify quercetin (QCT), using a simple carbon black paste electrode as a transducer. The proposed biosensor was characterized by electrochemical impedance spectroscopy and Nyquist plots were used to evaluate the immobilization of the enzyme. For determining QCT, variables were optimized, that included experimental conditions for laccase immobilization, pH of the supporting electrolyte, and instrumental parameters of the electroanalytical technique. From square-wave-voltammograms, a linear dependence between the cathodic current peak and QCT concentration was observed within the range 4.98–50.0 × 10−8 mol L−1, with a theoretical detection limit of 2.6 × 10−8 mol L−1. The proposed method was successfully applied to determine QCT in beverages, pharmaceuticals, and biological samples. The proposed biosensor device presented good selectivity in the presence of uric acid, various inorganic ions, as well as other phenolic compounds, demonstrating the potential application of this biosensing platform in chemically complex solutions. Operational and analytical stability of the laccase-biosensor were evaluated, and good intra-day (SD = 1.23%) and inter-day (SD = 2.32%) repeatability, and long storage stability (SD = 3.47%) are presented. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect