Abstract
Concentrations of deoxynivalenol (DON) and deepoxy deoxynivalenol (DOM-1) in animal blood are important parameters for studies in toxicology and biological detoxification of DON. Clean-up methods, using either immunoaffinity chromatography (IAC) or solid phase extraction (SPE), were compared in order to determine the free form of DON or DOM-1 and the sum amount (free form plus glucuronide conjugated form of DON or DOM-1), respectively, in swine serum. Detection was achieved by high performance liquid chromatography with ultraviolet detection (HPLC-UV). Compared with the SPE-HPLC method, the IAC-HPLC method provided lower quantitation limit (DON: 18 vs 42 ng/ml; DOM-1: 21 vs 30 ng/ml) and higher recoveries (DON: 93.4–102.7% vs 63.7–85.3%; DOM-1: 85.5–91.1% vs 68.0–82.6%). Compared with previously published methods, the developed IAC-HPLC method removed analytical interferences from swine serum in one quick and easy step, and eliminated steps of extraction with organic solvent and/or pre-purification using SPE cartridges. This IAC-HPLC method was used to analyze swine serum samples collected from pigs that were evaluated in a feeding trial of a microbiological detoxification of DON. No DON or DOM-1 were detected in serum samples from pigs given a toxin-free diet or a microbial control diet. In serum samples from pigs given a DON diet (5 mg/kg of DON), free form DON and sum free DON + conjugated DON were 38.8 ± 13.7 and 49.8 ± 14.1 ng/ml, respectively. In serum samples from those given a detoxified-DON diet (DON was transformed to DOM-1), free form DOM-1 was detected but not quantified, and the sum DOM-1 was found as 47.5 ± 6.3 ng/ml.
Similar content being viewed by others
References
Bily AC, Reid LM, Savard ME, Reddy R, Blackwell BA, Campbell CM, Krantis A, Durst T, Philogène BJR, Arnason JT, Regnault-Roger C (2004) Analysis of Fusarium graminearum mycotoxins in different biological matrices by LC/MS. Mycopathologia 157:117–126. doi:10.1023/B:MYCO.0000012218.27359.ec
Cahill LM, Kruger SC, McAlice BT, Ramsey CS, Prioli R, Kohn B (1999) Quantification of deoxynivalenol in wheat using an immunoaffinity column and liquid chromatography. J Chromatogr A 859:23–28. doi:10.1016/S0021-9673(99)00846-8
Cote LM, Beasley VR, Bratich PM, Swanson SP, Shivaprasad HL, Buck WB (1985) Sex-related reduced weight gains in growing swine fed diets containing deoxynivalenol. J Anim Sci 61:942–950
Dänicke S, Brüssow KP, Valenta H, Ueberschär KH, Tiemann U, Schollenberger M (2005) On the effects of graded levels of Fusarium toxin contaminated wheat in diets for gilts on feed intake, growth performance and metabolism of deoxynivalenol and zearalenone. Mol Nutr Food Res 49:932–943. doi:10.1002/mnfr.200500050
Döll S, Dänicke S, Ueberschär KH, Valenta H, Schnurrbusch U, Ganter M, Klobasa F, Flachowsky G (2003) Effects of graded levels of Fusarium toxin contaminated maize in diets for female weaned piglets. Arch Anim Nutr 57:311–334. doi:10.1080/00039420310001607680
Döll S, Goyarts T, Tiemann U, Dänicke S (2007) Practically relevant concentrations of deoxynivalenol in diets for growing-finishing pigs offered as mash or pellets. Arch Anim Nutr 61:247–265. doi:10.1080/17450390701431698
Eriksen GS, Pettersson H, Lindberg JE (2003) Absorption, metabolism and excretion of 3-acetyl don in pigs. Arch Anim Nutr 57:335–345. doi:10.1080/00039420310001607699
Federal Register (1995) “Definition and procedure for determination of the Method Detection Limit” 40 CFR Part 136, Appendix B, Revision 1.11 http://www.setonresourcecenter.com/CFR/40CFR/P136_008.HTM
Fuchs E, Binder EM, Heidler D, Krska R (2002) Structural characterization of metabolites after the microbial degradation of type A trichothecenes by the bacterial strain BBSH 797. Food Addit Contam 19:379–386. doi:10.1080/02652030110091154
Glaser JA, Foerst DL, McKee GD, Qyave SA, Budde WL (1981) Trace analyses for wastewaters. Environ Sci Technol 15:1426–1435. doi:10.1021/es00094a002
Goyarts T, Dänicke S (2006) Bioavailability of the Fusarium toxin deoxynivalenol (DON) from naturally contaminated wheat for the pig. Toxicol Lett 163:171–182. doi:10.1016/j.toxlet.2005.10.007
He J, Yang R, Zhou T, Tsao R, Young JC, Zhu H, Li X-Z, Boland GJ (2007) Purification of deoxynivalenol from Fusarium graminearum rice culture and mouldy corn by high-speed counter-current chromatography. J Chromatogr A 1151:187–192. doi:10.1016/j.chroma.2007.01.112
Hedman R, Pettersson H, Lindberg JE (1997) Absorption and metabolism of nivalenol in pigs. Arch Anim Nutr 50:13–24. doi:10.1080/17450399709386115
Janes W, Schuster M (2001) Determination of deoxynivalenol (DON) in blood, bile, urine and excrement samples from swine using immunoaffinity chromatography and LC-UV-detection. Mycotoxin Res 17:88–95. doi:10.1007/BF02946131
Lattanzio VMT, Solfrizzo M, Powers S, Visconti A (2007) Simultaneous determination of aflatoxins, ochratoxin A and Fusarium toxins in maize by liquid chromatography/tandem mass spectrometry after multitoxin immunoaffinity cleanup. Rapid Commun Mass Spectrom 21:3253–3261. doi:10.1002/rcm.3210
Li X-Z, Zhu J, de Lange CFM, Zhou T, He J, Yu H, Gong J, Zhu H, Young JC (2008) Microbial detoxification of deoxynivalenol (DON) demonstrated in a swine feeding trial. The 2008 CIFST Conference. Charlottetown, PEI, May 25–27, 2008. https://secure.cifst.ca/default.asp?id=1127
Nelson PE (2002) Fusarium-Paul E. Nelson Memorial Symposium. The American Phytopathological Society, St. Paul
Pestka JJ (2007) Deoxynivalenol: toxicity, mechanisms and animal health risks. Anim Feed Sci Technol 137:283–298. doi:10.1016/j.anifeedsci.2007.06.006
Pestka J, Smolinski A (2005) Deoxynivalenol: toxicology and potential effects on humans. J Toxicol Environ Health Part B 8:39–69
Prelusky DB, Trenholm HL (1991) Tissue distribution of deoxynivalenol in swine dosed intravenously. J Agric Food Chem 39:748–751. doi:10.1021/jf00004a026
Seeling K, Dänicke S, Valenta H, Van Egmond HP, Schothorst RC, Jekel AA, Lebzien P, Schollenberger M, Razzazi-Fazeli E, Flachowsky G (2006) Effects of Fusarium toxin-contaminated wheat and feed intake level on the biotransformation and carry-over of deoxynivalenol in dairy cows. Food Addit Contam 23:1008–1020. doi:10.1080/02652030600723245
Swanson SP, Terwel L, Corley RA, Buck WB (1982) Gas chromatographic method for the determination of diacetoxyscirpenol in swine plasma and urine. J Chromatogr A 248:456–460. doi:10.1016/S0021-9673(00)85057-8
Valenta H, Dänicke S (2005) Study on the transmission of deoxynivalenol and deepoxy-deoxynivalenol into eggs of laying hens using a high-performance liquid chromatography-ultraviolet method with clean-up by immunoaffinity columns. Mol Nutr Food Res 49:779–785. doi:10.1002/mnfr.200500012
Valenta H, Dänicke S, Döll S (2003) Analysis of deoxynivalenol and deepoxy-deoxynivalenol in animal tissue by liquid chromatography after clean-up with an immunoaffinity column. Mycotoxin Res 19:51–55. doi:10.1007/BF02940093
Wu X, Murphy P, Cunnick J, Hendrich S (2007) Synthesis and characterization of deoxynivalenol glucuronide: its comparative immunotoxicity with deoxynivalenol. Food Chem Toxicol 45:1846–1855. doi:10.1016/j.fct.2007.03.018
Young JC, Zhou T, Yu H, Zhu H, Gong J (2007) Degradation of trichothecene mycotoxins by chicken intestinal microbes. Food Chem Toxicol 45:136–143. doi:10.1016/j.fct.2006.07.028
Acknowledgements
The authors thank Julia Zhu from the University of Guelph and Honghui Zhu from the AAFC for their technical assistance.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
He, J., Li, XZ. & Zhou, T. Sample clean-up methods, immunoaffinity chromatography and solid phase extraction, for determination of deoxynivalenol and deepoxy deoxynivalenol in swine serum. Mycotox Res 25, 89–94 (2009). https://doi.org/10.1007/s12550-009-0013-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12550-009-0013-3