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Electrochemical biosensors with spatially separated enzymatic and detection parts for selective analysis in flow-through arrangement
Tvorynska, Sofiia ; Barek, Jiří (advisor) ; Labuda, Ján (referee) ; Korecká, Lucie (referee)
This dissertation thesis presents the newly developed four highly reusable, stable as well as simple, and cost-effective electrochemical (bi)enzymatic biosensors for the selective and reliable determination of choline, acetylcholine, uric acid, and L-lactic acid in flow injection analysis. All biosensors are based on the concept of the spatial separation of the biorecognition part from detection one and amperometric monitoring of the enzymatically consumed oxygen via its four-electron reduction at the highly negative detection potential. In this way, the design of the biosensors includes an easily replaceable enzymatic mini-reactor(s) connected upstream to the flow cell that contains the appropriate silver amalgam-based transducer. The enzymatic mini-reactor based on choline oxidase, uricase, or lactate oxidase was used for choline, uric acid, or L-lactic acid biosensors, respectively. The acetylcholine bienzymatic biosensor includes the consequently connected choline oxidase- and acetylcholinesterase-based mini-reactors. The first part of this thesis focuses on the construction of two different silver amalgam-based electrodes. Specifically, this section discusses the fabrication of a silver solid amalgam electrode covered by mercury film operating in a wall-jet cell and also highlights the...
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Construction and Application of Flow Amperometric Biosensor Based on Enzymatic Reactor for Determination of Choline
Tvorynska, Sofiia ; Barek, J. ; Josypčuk, Bohdan
In this contribution, a novel fast, simple and stable biosensor with the enzymatic reactor based on choline oxidase (ChOx) was developed and applied for the determination of choline using flow injection analysis with amperometric detection. The preparation of the reactor is based on the covalent immobilization of ChOx with glutaraldehyde to mesoporous silica powder (SBA-15) previously covered by NH2-groups. The experimental parameters affecting the sensitivity and stability of the biosensor were optimized. The proposed biosensor with the newly developed ChOx-based reactor possesses good repeatability, reproducibility, long-term stability, and reusability. It was successfully applied for the determination of choline in commercial pharmaceuticals.
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