Optimization of determination of free and total deoxynivalenol in urine

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F22%3A43925472" target="_blank" >RIV/60461373:22330/22:43925472 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Optimization of determination of free and total deoxynivalenol in urine

Popis výsledku v původním jazyce

Deoxynivalenol (DON) and other structurally similar trichothecene mycotoxins are rapidly metabolized and excreted in humans. Despite this fact, DON shows strong emetic effects and from the view of chronic effects also pose a risk due to protein synthesis inhibition and cytotoxicity. Therefore it is regulated in EU and many other countries worldwide and European Food Safety Authority (EFSA) set a tolerable daily intake (TDI) for DON at 1 μg/kg body weight/day. DON is predominantly excreted in urine mainly after glucuronidation as DON-3-glucuronide, DON-15-glucuronide and to a lesser extent in a free form. Detection of other minor human urinary metabolites of DON has been also described recently. As regards analytical determination of total DON in urine, e.g. free DON and DON associated in DON-glucuronides, enzymatic deconjugation using beta-glucuronidase is typically used in a combination with clean-up step employing immunoaffinity columns (IAC) and liquid chromatography coupled with mass spectrometric detection (LC−MS). In this study, (i) excretion rate of free and glucuronidated DON was evaluated within a small scale study using artificial contamination of a beverage to the TDI level of a volunteer followed by (ii) optimization of enzymatic hydrolysis of DON-glucuronides (hydrolysis pH, temperature, time and number of enzyme units) contained in urine using beta-glucuronidase from Helix pomatia and subsequent (iii) instrumental analysis optimization (ionization and detection parameters) using ultrahigh performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC−HRMS/MS) employing Q-Exactive Plus. The aim of the study was to achieve low limit of quantification (LOQ) without the need utilize time consuming and costly IAC approach which, on the other hand, allows to obtain practically arbitrarily low LOQ depending on the amount of sample pushed through the IAC cartridge. Our approach resulted in acceptable LOQ of 1 μg/L.

Název v anglickém jazyce

Optimization of determination of free and total deoxynivalenol in urine

Popis výsledku anglicky

Deoxynivalenol (DON) and other structurally similar trichothecene mycotoxins are rapidly metabolized and excreted in humans. Despite this fact, DON shows strong emetic effects and from the view of chronic effects also pose a risk due to protein synthesis inhibition and cytotoxicity. Therefore it is regulated in EU and many other countries worldwide and European Food Safety Authority (EFSA) set a tolerable daily intake (TDI) for DON at 1 μg/kg body weight/day. DON is predominantly excreted in urine mainly after glucuronidation as DON-3-glucuronide, DON-15-glucuronide and to a lesser extent in a free form. Detection of other minor human urinary metabolites of DON has been also described recently. As regards analytical determination of total DON in urine, e.g. free DON and DON associated in DON-glucuronides, enzymatic deconjugation using beta-glucuronidase is typically used in a combination with clean-up step employing immunoaffinity columns (IAC) and liquid chromatography coupled with mass spectrometric detection (LC−MS). In this study, (i) excretion rate of free and glucuronidated DON was evaluated within a small scale study using artificial contamination of a beverage to the TDI level of a volunteer followed by (ii) optimization of enzymatic hydrolysis of DON-glucuronides (hydrolysis pH, temperature, time and number of enzyme units) contained in urine using beta-glucuronidase from Helix pomatia and subsequent (iii) instrumental analysis optimization (ionization and detection parameters) using ultrahigh performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC−HRMS/MS) employing Q-Exactive Plus. The aim of the study was to achieve low limit of quantification (LOQ) without the need utilize time consuming and costly IAC approach which, on the other hand, allows to obtain practically arbitrarily low LOQ depending on the amount of sample pushed through the IAC cartridge. Our approach resulted in acceptable LOQ of 1 μg/L.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

30304 - Public and environmental health

Projekt

<a href="/cs/project/LM2018100" target="_blank" >LM2018100: Infrastruktura pro propagaci metrologie v potravinářství a výživě v České republice</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2022

Kód důvěrnosti údajů

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