Hyaluronidase reaction kinetics evaluated by capillary electrophoresis with UV and high-resolution mass spectrometry (HRMS) detection

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F17%3A00094585" target="_blank" >RIV/00216224:14310/17:00094585 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.aca.2016.11.036" target="_blank" >http://dx.doi.org/10.1016/j.aca.2016.11.036</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.aca.2016.11.036" target="_blank" >10.1016/j.aca.2016.11.036</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Hyaluronidase reaction kinetics evaluated by capillary electrophoresis with UV and high-resolution mass spectrometry (HRMS) detection

Popis výsledku v původním jazyce

A capillary electrophoresis (CE) method with enzymatic reaction inside the capillary was developed for the study of hyaluronidase activity. Capillary electrophoresis was used for the first time for the determination of the kinetic constants (Km, Vmax and IC50) of the enzymatic degradation of hyaluronic acid. Km and Vmax were determined using CE with UV and MS detection. Reaction products were identified using CE-MS. IC50 of a standard natural inhibitor, epigallocatechin gallate, was also determined by CE-UV and CE-MS. Kinetic constant values obtained by CE compared well with literature which validated the developed CE-based assay. Homemade oligosaccharides were tested to search for new hyaluronidase inhibitors (or activators). Sulfatation in 6- and 4- position on chondroitin tetrasacharides were found to inhibit hydrolysis of hyaluronic acid. Small modification of one hydroxyl group in a trisaccharide compound was shown to be enough for changing effect on hyaluronidase activity. CE-based assay can be used for the screening of natural and synthetic inhibitors of hyaluronidase activity for cosmetic and therapeutic applications.

Název v anglickém jazyce

Hyaluronidase reaction kinetics evaluated by capillary electrophoresis with UV and high-resolution mass spectrometry (HRMS) detection

Popis výsledku anglicky

A capillary electrophoresis (CE) method with enzymatic reaction inside the capillary was developed for the study of hyaluronidase activity. Capillary electrophoresis was used for the first time for the determination of the kinetic constants (Km, Vmax and IC50) of the enzymatic degradation of hyaluronic acid. Km and Vmax were determined using CE with UV and MS detection. Reaction products were identified using CE-MS. IC50 of a standard natural inhibitor, epigallocatechin gallate, was also determined by CE-UV and CE-MS. Kinetic constant values obtained by CE compared well with literature which validated the developed CE-based assay. Homemade oligosaccharides were tested to search for new hyaluronidase inhibitors (or activators). Sulfatation in 6- and 4- position on chondroitin tetrasacharides were found to inhibit hydrolysis of hyaluronic acid. Small modification of one hydroxyl group in a trisaccharide compound was shown to be enough for changing effect on hyaluronidase activity. CE-based assay can be used for the screening of natural and synthetic inhibitors of hyaluronidase activity for cosmetic and therapeutic applications.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10600 - Biological sciences

Projekt

<a href="/cs/project/GBP206%2F12%2FG014" target="_blank" >GBP206/12/G014: Centrum pokročilých bioanalytických technologií</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2017

Kód důvěrnosti údajů

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

Název periodika

Analytica Chimica Acta

ISSN

0003-2670

e-ISSN

—

Svazek periodika

951

Číslo periodika v rámci svazku

January

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

11

Strana od-do

140-150

Kód UT WoS článku

000392773800011

EID výsledku v databázi Scopus

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