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Liquid chromatographic methods for biotransformation studies of ochratoxin A

Annelore Schaut, Sarah De Saeger UGent, T Sergent, YJ Schneider, Y Larondelle, L Pussemier, R Blank and Carlos Van Peteghem UGent (2008) BIOMEDICAL CHROMATOGRAPHY. 22(9). p.1013-1020
abstract
Liquid chromatographic methods were used for the detection of ochratoxin A (OTA) and its metabolites ochratoxin alpha (OT alpha), 10-hydroxy OTA (10-OHOTA), 4R-hydroxy OTA (4R-OHOTA) and the ethyl ester of OTA (OTC) in in vitro samples, obtained with Caco-2 cell culture experiments and in in vivo urine samples from sheep. A high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method were developed and validated for the detection of OTA and its metabolites OTo 10-OHOTA, 4R-OHOTA and OTC, which was used as internal standard. The LOD/LOQ values for OT alpha, 4R-OHOTA and OTA were 0.63/2.11, 0.99/3.31 and 0.84/2.81 mu g/L, respectively, for the HPLC-FLD method and 0.98/3.28, 1.11/3.72 and 0.88/2.96 mu g/L, respectively for the LC-MS/MS method. Within-day and between-day precision were both <12% for the HPLC-FLD method, and <10% for the LC-MS/MS method. The recovery of OTA and its metabolites ranged between 71 and 111% for the HPLC-FLD method and between 79 and 110 % for the LC-MS/MS method. In the first experiment only OTA was added to the Caco-2 cells while in the second experiment 3-methylcholanthrene (3MC) was also present in the cell culture systems. Besides OTA, which was recovered in all the samples, an unknown compound was also observed in the second experiment. When 3MC was added, the results showed that the OTA concentration in the basolateral samples was decreased by 50%. The methods were also implemented for the analysis of urine samples of sheep, fed increasing amounts of OTA. With the HPLC-FLD method it could be concluded that the concentration of OTA and OTa increased according to ingested amounts of OTA, with OTa being the most abundant compound. The results obtained with the LC-MS/MS method confirmed these results.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
CELLS, METABOLITES, ASPERGILLUS-OCHRACEUS, ABSORPTION, MYCOTOXIN, HYDROLYSIS, TOXICOKINETICS, RATS, CONVERSION, EXCRETION
journal title
BIOMEDICAL CHROMATOGRAPHY
Biomed. Chromatogr.
editor
Marianne Bailleul UGent
volume
22
issue
9
pages
1013 - 1020
Web of Science type
Article
Web of Science id
000259616600011
JCR category
CHEMISTRY, ANALYTICAL
JCR impact factor
1.592 (2008)
JCR rank
39/70 (2008)
JCR quartile
3 (2008)
ISSN
0269-3879
DOI
10.1002/bmc.1021
language
English
UGent publication?
yes
classification
A1
id
531275
handle
http://hdl.handle.net/1854/LU-531275
date created
2009-03-26 10:56:55
date last changed
2009-03-31 08:44:30
@article{531275,
  abstract     = {Liquid chromatographic methods were used for the detection of ochratoxin A (OTA) and its metabolites ochratoxin alpha (OT alpha), 10-hydroxy OTA (10-OHOTA), 4R-hydroxy OTA (4R-OHOTA) and the ethyl ester of OTA (OTC) in in vitro samples, obtained with Caco-2 cell culture experiments and in in vivo urine samples from sheep. A high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method were developed and validated for the detection of OTA and its metabolites OTo 10-OHOTA, 4R-OHOTA and OTC, which was used as internal standard. The LOD/LOQ values for OT alpha, 4R-OHOTA and OTA were 0.63/2.11, 0.99/3.31 and 0.84/2.81 mu g/L, respectively, for the HPLC-FLD method and 0.98/3.28, 1.11/3.72 and 0.88/2.96 mu g/L, respectively for the LC-MS/MS method. Within-day and between-day precision were both {\textlangle}12\% for the HPLC-FLD method, and {\textlangle}10\% for the LC-MS/MS method. The recovery of OTA and its metabolites ranged between 71 and 111\% for the HPLC-FLD method and between 79 and 110 \% for the LC-MS/MS method. In the first experiment only OTA was added to the Caco-2 cells while in the second experiment 3-methylcholanthrene (3MC) was also present in the cell culture systems. Besides OTA, which was recovered in all the samples, an unknown compound was also observed in the second experiment. When 3MC was added, the results showed that the OTA concentration in the basolateral samples was decreased by 50\%. The methods were also implemented for the analysis of urine samples of sheep, fed increasing amounts of OTA. With the HPLC-FLD method it could be concluded that the concentration of OTA and OTa increased according to ingested amounts of OTA, with OTa being the most abundant compound. The results obtained with the LC-MS/MS method confirmed these results.},
  author       = {Schaut, Annelore and De Saeger, Sarah and Sergent, T and Schneider, YJ and Larondelle, Y and Pussemier, L and Blank, R and Van Peteghem, Carlos},
  editor       = {Bailleul, Marianne},
  issn         = {0269-3879},
  journal      = {BIOMEDICAL CHROMATOGRAPHY},
  keyword      = {CELLS,METABOLITES,ASPERGILLUS-OCHRACEUS,ABSORPTION,MYCOTOXIN,HYDROLYSIS,TOXICOKINETICS,RATS,CONVERSION,EXCRETION},
  language     = {eng},
  number       = {9},
  pages        = {1013--1020},
  title        = {Liquid chromatographic methods for biotransformation studies of ochratoxin A},
  url          = {http://dx.doi.org/10.1002/bmc.1021},
  volume       = {22},
  year         = {2008},
}

Chicago
Schaut, Annelore, Sarah De Saeger, T Sergent, YJ Schneider, Y Larondelle, L Pussemier, R Blank, and Carlos Van Peteghem. 2008. “Liquid Chromatographic Methods for Biotransformation Studies of Ochratoxin A.” Ed. Marianne Bailleul. Biomedical Chromatography 22 (9): 1013–1020.
APA
Schaut, A., De Saeger, S., Sergent, T., Schneider, Y., Larondelle, Y., Pussemier, L., Blank, R., et al. (2008). Liquid chromatographic methods for biotransformation studies of ochratoxin A. (M. Bailleul, Ed.)BIOMEDICAL CHROMATOGRAPHY, 22(9), 1013–1020.
Vancouver
1.
Schaut A, De Saeger S, Sergent T, Schneider Y, Larondelle Y, Pussemier L, et al. Liquid chromatographic methods for biotransformation studies of ochratoxin A. Bailleul M, editor. BIOMEDICAL CHROMATOGRAPHY. 2008;22(9):1013–20.
MLA
Schaut, Annelore, Sarah De Saeger, T Sergent, et al. “Liquid Chromatographic Methods for Biotransformation Studies of Ochratoxin A.” Ed. Marianne Bailleul. BIOMEDICAL CHROMATOGRAPHY 22.9 (2008): 1013–1020. Print.