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Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods | DIAL.pr - BOREAL

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Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods

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Schaut, A. De Saeger, S. Sergent, Thérèse [UCL] Schneider, Yves-Jacques [UCL] Larondelle, Yvan [UCL] Van Peteghem, Carlos

A high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) was developed and validated for the detection of zearalenone (ZON), alpha-zearalenon (alpha-ZOL) and beta-zearalenol (beta-ZOL) in in vitro biological samples. Furthermore, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the detection of ZON, alpha-ZOL, beta-ZOL, alpha-zearalanol (alpha-ZAL) and beta-zearalanol (beta-ZAL) in in vitro biological samples. Zearalanone (ZAN) was used as internal standard in both methods. The limit of detection/limit of quantitation (LOD/LOQ) values of ZON, alpha-ZOL and beta-ZOL were 2/7, 2/7 and 4/13 mu g 1(-1), respectively, for the HPLC-FLD method. For the LC-MS/MS method LOD/LOQ values for ZON, alpha-ZOL, beta-ZOL, alpha-ZAL and beta-ZAL were 6/20, 5/17, 4/14, 9/30 and 6/19 mu g 1(-1), respectively. Within-day and between-day precision were less then 11 and 14%, respectively for the HPLC-FLD method, and both less then 20% for the LC-MS/MS method. The recovery of ZON and its metabolities ranged between 73 and 89% for the HPLC-FLD method and between 69 and 112% for the LC-MS/MS method. The methods were used for the detection of the compounds in in vitro biological samples, obtained with human intestinal Caco-2 cells culture experiments. The 8-days post-confluent Caco-2 cells were treated with ZON or a mixture of ZON and imazalil (IMA). After an incubation time of 24 h the samples were analyzed with the HPLC-FLD method. Neither ZON nor its derivatives were detected in the samples. The disappearance of ZON could possibly point out the formation of phase II metabolities like glucuronide conjugates. There, samples were pretreated with beta-glucuronidase before LC-MS/MS analysis. The LC-MS/MS results showed that ZON, alpha-ZOL and beta-ZOL could only be detected in the beta-glucuronidase pretreated samples. This confirmed the formation of glucuronide conjugates and the hydroxylation of ZON during the incubation with Caco-2 cells. Copyright (C) 2008 John Wiley & Sons, Ltd.

Document type Article de périodique (Journal article) – Article de recherche
Publication date 2008
Language Anglais
Journal information "Journal of Applied Toxicology" - Vol. 28, no. 8, p. 966-973 (2008)
Peer reviewed yes
Publisher John Wiley & Sons Ltd (Chichester)
issn 0260-437X
e-issn 1099-1263
Publication status Publié
Affiliations UCL - SC/CHIM - Département de chimie
UCL - AGRO/BAPA - Département de biologie appliquée et des productions agricoles
Keywords zearalenone ; biotransformation ; Caco-2 ; imazalil ; analytical methodology ; glucuronidation
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Bibliographic reference Schaut, A. ; De Saeger, S. ; Sergent, Thérèse ; Schneider, Yves-Jacques ; Larondelle, Yvan ; et. al. Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods. In: Journal of Applied Toxicology, Vol. 28, no. 8, p. 966-973 (2008)
Permanent URL http://hdl.handle.net/2078.1/36220