Offline and online capillary electrophoresis enzyme assays of beta-N-acetylhexosaminidase

Kód výsledku v IS VaVaI

RIV/00216208:11310/13:10133390 - isvavai.cz

Výsledek na webu

http://dx.doi.org/10.1007/s00216-012-6607-1

DOI - Digital Object Identifier

10.1007/s00216-012-6607-1

Jazyk výsledku

angličtina

Název v původním jazyce

Offline and online capillary electrophoresis enzyme assays of beta-N-acetylhexosaminidase

Popis výsledku v původním jazyce

Enzyme assays of beta-N-acetylhexosaminidase from Aspergillus oryzae using capillary electrophoresis in the offline and online setup have been developed. The pH value and concentration of the borate-based background electrolyte were optimized in order toachieve baseline separation of N,N',N"-triacetylchitotriose, N,N'-diacetylchitobiose, and N-acetyl-D-glucosamine. The optimized method using 25 mM tetraborate buffer, pH 10.0, was evaluated in terms of repeatability, limits of detection, quantification,and linearity. The method was successfully applied to the offline enzyme assay of beta-N-acetylhexosaminidase, which was demonstrated by monitoring the hydrolysis of N,N',N"- triacetylchitotriose. The presented method was also utilized to study the pH dependence of enzyme activity. An online assay with N,N'-diacetylchitobiose as a substrate was developed using the Transverse Diffusion of Laminar Flow Profiles model to optimize the injection sequence and incapillary mixing of substrate a

Název v anglickém jazyce

Offline and online capillary electrophoresis enzyme assays of beta-N-acetylhexosaminidase

Popis výsledku anglicky

Enzyme assays of beta-N-acetylhexosaminidase from Aspergillus oryzae using capillary electrophoresis in the offline and online setup have been developed. The pH value and concentration of the borate-based background electrolyte were optimized in order toachieve baseline separation of N,N',N"-triacetylchitotriose, N,N'-diacetylchitobiose, and N-acetyl-D-glucosamine. The optimized method using 25 mM tetraborate buffer, pH 10.0, was evaluated in terms of repeatability, limits of detection, quantification,and linearity. The method was successfully applied to the offline enzyme assay of beta-N-acetylhexosaminidase, which was demonstrated by monitoring the hydrolysis of N,N',N"- triacetylchitotriose. The presented method was also utilized to study the pH dependence of enzyme activity. An online assay with N,N'-diacetylchitobiose as a substrate was developed using the Transverse Diffusion of Laminar Flow Profiles model to optimize the injection sequence and incapillary mixing of substrate a

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

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Projekt

GBP206/12/G151: Centrum nových přístupů k bioanalýze a molekulární diagnostice

Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)
S - Specificky vyzkum na vysokych skolach
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2013

Kód důvěrnosti údajů

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

Název periodika

Analytical and Bioanalytical Chemistry

ISSN

1618-2642

e-ISSN

—

Svazek periodika

405

Číslo periodika v rámci svazku

8

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

10

Strana od-do

2425-2434

Kód UT WoS článku

000315464800003

EID výsledku v databázi Scopus

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