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

(2008) JOURNAL OF APPLIED TOXICOLOGY. 28(8). p.966-973
Author
Organization
Abstract
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.
Keywords
analytical methodology, biotransformation, zearalenone, imazalil, Caco-2, glucuronidation

Citation

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MLA
Schaut, Annelore, et al. “Study of the Gastrointestinal Biotransformation of Zearalenone in a Caco-2 Cell Culture System with Liquid Chromatographic Methods.” JOURNAL OF APPLIED TOXICOLOGY, vol. 28, no. 8, 2008, pp. 966–73, doi:10.1002/jat.1362.
APA
Schaut, A., De Saeger, S., Sergent, T., Schneider, Y., Larondelle, Y., Pussemier, L., & Van Peteghem, C. (2008). Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods. JOURNAL OF APPLIED TOXICOLOGY, 28(8), 966–973. https://doi.org/10.1002/jat.1362
Chicago author-date
Schaut, Annelore, Sarah De Saeger, T. Sergent, Yves Schneider, Yvan Larondelle, Luc Pussemier, and Carlos Van Peteghem. 2008. “Study of the Gastrointestinal Biotransformation of Zearalenone in a Caco-2 Cell Culture System with Liquid Chromatographic Methods.” JOURNAL OF APPLIED TOXICOLOGY 28 (8): 966–73. https://doi.org/10.1002/jat.1362.
Chicago author-date (all authors)
Schaut, Annelore, Sarah De Saeger, T. Sergent, Yves Schneider, Yvan Larondelle, Luc Pussemier, and Carlos Van Peteghem. 2008. “Study of the Gastrointestinal Biotransformation of Zearalenone in a Caco-2 Cell Culture System with Liquid Chromatographic Methods.” JOURNAL OF APPLIED TOXICOLOGY 28 (8): 966–973. doi:10.1002/jat.1362.
Vancouver
1.
Schaut A, De Saeger S, Sergent T, Schneider Y, Larondelle Y, Pussemier L, et al. Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods. JOURNAL OF APPLIED TOXICOLOGY. 2008;28(8):966–73.
IEEE
[1]
A. Schaut et al., “Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods,” JOURNAL OF APPLIED TOXICOLOGY, vol. 28, no. 8, pp. 966–973, 2008.
@article{680923,
  abstract     = {{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.}},
  author       = {{Schaut, Annelore and De Saeger, Sarah and Sergent, T. and Schneider, Yves and Larondelle, Yvan and Pussemier, Luc and Van Peteghem, Carlos}},
  issn         = {{0260-437X}},
  journal      = {{JOURNAL OF APPLIED TOXICOLOGY}},
  keywords     = {{analytical methodology,biotransformation,zearalenone,imazalil,Caco-2,glucuronidation}},
  language     = {{eng}},
  number       = {{8}},
  pages        = {{966--973}},
  title        = {{Study of the gastrointestinal biotransformation of zearalenone in a Caco-2 cell culture system with liquid chromatographic methods}},
  url          = {{http://doi.org/10.1002/jat.1362}},
  volume       = {{28}},
  year         = {{2008}},
}

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