Amperometric Biosensor Based on Enzymatic Reactor for Choline Determination in Flow Systems

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388955%3A_____%2F19%3A00510097" target="_blank" >RIV/61388955:_____/19:00510097 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/19:10402080

Výsledek na webu

<a href="http://hdl.handle.net/11104/0300653" target="_blank" >http://hdl.handle.net/11104/0300653</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/elan.201900237" target="_blank" >10.1002/elan.201900237</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Amperometric Biosensor Based on Enzymatic Reactor for Choline Determination in Flow Systems

Popis výsledku v původním jazyce

A simple, selective and stable biosensor with the enzymatic reactor based on choline oxidase (ChOx) was developed and applied for the determination of choline (Ch) in flow injection analysis with amperometric detection. The enzyme ChOx was covalently immobilized with glutaraldehyde to mesoporous silica powder (SBA-15) previously covered by NH2-groups. This powder was found as an optimal filling of the reactor. The detection of Ch is based on amperometric monitoring of consumed oxygen during the enzymatic reaction, which is directly proportional to Ch concentration. Two arrangements of an electrolytic cell in FIA, namely wall-jet cell with working silver solid amalgam electrode covered by mercury film and flow-through cell with tubular detector of polished silver solid amalgam were compared. The experimental parameters affecting the sensitivity and stability of the biosensor (i. e. pH of the carrier solution, volume of reactor, amount of the immobilized enzyme, the detection potential, flow rate, etc.) were optimized. Under the optimized conditions, the limit of detection was found to be 9.0×10−6 mol L−1. The Michaelis-Menten constant for covalently immobilized ChOx on SBA-15 was calculated. The proposed amperometric biosensor with the developed ChOx-based reactor exhibits good repeatability, reproducibility, long-term stability, and reusability. Its efficiency has been confirmed by the successful application for the determination of Ch in two commercial pharmaceuticals.

Název v anglickém jazyce

Amperometric Biosensor Based on Enzymatic Reactor for Choline Determination in Flow Systems

Popis výsledku anglicky

A simple, selective and stable biosensor with the enzymatic reactor based on choline oxidase (ChOx) was developed and applied for the determination of choline (Ch) in flow injection analysis with amperometric detection. The enzyme ChOx was covalently immobilized with glutaraldehyde to mesoporous silica powder (SBA-15) previously covered by NH2-groups. This powder was found as an optimal filling of the reactor. The detection of Ch is based on amperometric monitoring of consumed oxygen during the enzymatic reaction, which is directly proportional to Ch concentration. Two arrangements of an electrolytic cell in FIA, namely wall-jet cell with working silver solid amalgam electrode covered by mercury film and flow-through cell with tubular detector of polished silver solid amalgam were compared. The experimental parameters affecting the sensitivity and stability of the biosensor (i. e. pH of the carrier solution, volume of reactor, amount of the immobilized enzyme, the detection potential, flow rate, etc.) were optimized. Under the optimized conditions, the limit of detection was found to be 9.0×10−6 mol L−1. The Michaelis-Menten constant for covalently immobilized ChOx on SBA-15 was calculated. The proposed amperometric biosensor with the developed ChOx-based reactor exhibits good repeatability, reproducibility, long-term stability, and reusability. Its efficiency has been confirmed by the successful application for the determination of Ch in two commercial pharmaceuticals.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10403 - Physical chemistry

Projekt

<a href="/cs/project/GA17-05387S" target="_blank" >GA17-05387S: Nové přístupy pro sledování přenosu zvolených xenobiotik přes biomimetické membrány</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Electroanalysis

ISSN

1040-0397

e-ISSN

—

Svazek periodika

31

Číslo periodika v rámci svazku

10

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

12

Strana od-do

1901-1912

Kód UT WoS článku

000491476600009

EID výsledku v databázi Scopus

2-s2.0-85067473082