Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
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Rapid and simultaneous purification of aflatoxin B1, zearalenone and deoxynivalenol using their monoclonal antibodies and magnetic nanoparticles

  • Received : 2020.09.28
  • Accepted : 2020.12.17
  • Published : 2021.10.15
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Abstract

To develop a new simple and simultaneous purification method for mycotoxins in feeds and grains, magnetic nanoparticles (MNPs) conjugated with monoclonal antibodies (mAbs) against mycotoxins were used to separate aflatoxin B1 (AFB1), zearalenone (ZEA) and deoxynivalenol (DON). For a single spike of each mycotoxin into the buffer solution (16% MeOH in PBS), mean recoveries were 93.1-95.0% for AFB1 (5-20 ng/mL spiked), 87.2-96.0% for ZEA (125-500 ng/mL spiked) and 75.2-96.9% for DON (250-1,000 ng/mL spiked) by HPLC and ELISA. Recoveries of AFB1 (20 ng/mL) and ZEA (500 ng/mL) simultaneously spiked into the buffer solution were 87.0 and 99.8%, respectively. Recovery rates of AFB1/DON and DON/ZEA spiked simultaneously were 86.2%/76.6% and 92.0%/86.7%, respectively, at concentrations of 20 ng/mL AFB1, 500 ng/mL ZEA, and 1,000 ng/mL DON. Recoveries using the novel mAb-MNP conjugated system in a buffer solution simultaneously spiked with AFB1, ZEA and DON were 82.5, 94.6 and 73.4%, respectively. Recoveries of DON in animal feed were 107.7-132.5% at concentrations of 250-1,000 ng/g spiked in feed. The immunoaffinity chromatography (IAC) clean-up method was compared with the purification method using novel mAb-MNP. After fortification of animal feed with AFB1 (5, 10 and 20 ng/g feed) and ZEA (125, 250 and 500 ng/g feed), AFB1 and ZEA were purified using both the methods. In the case of the novel mAb-MNP conjugated system, mean recoveries for AFB1 were 89.4, 73.1 and 88.3% at concentrations of 5, 10 and 20 ng/g feed, respectively. For ZEA, mean recoveries were 86.7, 85.9 and 79.1% at concentrations of 125, 250 and 500 ng/g, respectively. For IAC purification, recoveries were 42.9-45.1% for AFB1 and 96.8-103.2% for ZEA. In conclusion, the present purification method using monoclonal antibodies conjugated to MNPs can be used for simple and simultaneous purification of mycotoxins from feed and maize.

Keywords

Acknowledgement

This study was conducted as an international collaborative research endeavour between the National Veterinary Research and Quarantine Service (NVRQS) in the Republic of Korea and the Department of Population Medicine and Diagnostic Sciences of Cornell University in the U.S. and funded by the NVRQS (I-AD-2008-11-01). This study was also support by the research fund from Semyung University.

References

  1. Orti DL, Hill RH, Liddle JA, Needham LL, Vickers L (1986) High performance liquid chromatography of mycotoxin metabolites in human urine. J Anal Toxiol 10:41-45. https://doi.org/10.1093/jat/10.2.41
  2. Brezina U, Valenta H, Rempe I, Kersten S, Hans- Humpf HU, Danicke S (2014) Development of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of zearalenone, deoxynivalenol and their metabolites in pig serum. Mycotoxin Res 30:171-186. https://doi.org/10.1007/s12550-014-0200-8
  3. Scott PM, Trucksess MW (1997) Application of immunoaffinity columns to mycotoxin analysis. J AOAC Int 80:941-949. https://doi.org/10.1093/jaoac/80.5.941
  4. Takino M, Tanaka H, Tanaka T (2011) Multi mycotoxin analysis in food products using immunoaffinity extraction. Methods Mol Biol 747:259-266. https://doi.org/10.1007/978-1-61779-136-9_11
  5. Napolitano R, Soesbe TC, De Leon-Rodriguez LM, Sherry AD, Udugamasooriya DG (2011) On-bead combinatorial synthesis and imaging of chemical exchange saturation transfer magnetic resonance imaging agents to identify factors that influence water exchange. J Am Chem Soc 133:13023-13030. https://doi.org/10.1021/ja201123f
  6. Nash MA, Yager P, Hoffman AS, Stayton PS (2010) Mixed stimuli-responsive magnetic and gold nanop system for rapid purification, enrichment, and detection of biomarkers. Bioconjugate Chem 21:2197-2204. https://doi.org/10.1021/bc100180q
  7. Varshney M, Yang L, Su XL, Li Y (2005) Magnetic nanop-antibody conjugates for the separation of Escherichia coli O157:H7 in ground beef. J Food Protect 68:1804-1811. https://doi.org/10.4315/0362-028X-68.9.1804
  8. Wang L, Li Y, Mustaphai A (2007) Rapid and simultaneous quantitation of Escherichia coli 0157:H7, Salmonella, and Shigella in ground beef by multiplex real-time PCR and immunomagnetic separation. J Food Protect 70:1366-1372. https://doi.org/10.4315/0362-028X-70.6.1366
  9. Yoshitomi KJ, Jinneman KC, Zapata R, Weagant SD, Fedio WM (2012) Detection and isolation of low levels of E. coli O157:H7 in cilantro by real-time PCR, immunomagnetic separation, and cultural methods with and without an acid treatment. J Food Sci 77:M481-M489. https://doi.org/10.1111/j.1750-3841.2012.02813.x
  10. Lee HM, Song SO, Cha SH, Wee SB, Bischoff K, Park SW, Son SW, Kang HG, Cho MH (2013) Development of a monoclonal antibody against deoxynivalenol for magnetic nanop-based extraction and an enzyme-linked immunosorbent assay. J Vet Sci 14:143-150. https://doi.org/10.4142/jvs.2013.14.2.143
  11. Kim HJ, Kim SH, Lee JK, Choi CU, Lee HS, Kang HG, Cha SH (2012) A novel mycotoxin purification system using magnetic nanops for the recovery of aflatoxin B1 and zearalenone from feed. J Vet Sci 13:363-369. https://doi.org/10.4142/jvs.2012.13.4.363
  12. Alexa E, Dehelean CA, Poiana MA, Radulov I, Cimpean AM, Bordean DM, Tulcan C, Pop G (2013) The occurrence of mycotoxins in wheat from western Romania and histopathological impact as effect of feed intake. Chem Cent J 7:99. https://doi.org/10.1186/1752-153x-7-99
  13. Duarte SC, Lino CM, Pena A (2010) Mycotoxin food and feed regulation and the specific e of ochratoxin A: a review of the worldwide status. Food Addit Contam 27:1440-1450. https://doi.org/10.1080/19440049.2010.497166
  14. Zmudzki J, Wisniewska-Dmytrow H (2004) Limits and regulations for mycotoxins in food and feed. Polish J Vet Sci 7:211-216. https://.ncbi.nlm.nih.gov/15478869/
  15. Aqai P, Peters J, Gerssen A, Haasnoot W, Nielen MW (2011) Immunomagnetic microbeads for screening with flow cytometry and identification with nano-liquid chromatography mass spectrometry of ochratoxins in wheat and cereal. Anal Bioanal Chem 400:3085-3096. https://doi.org/10.1007/s00216-011-4974-7
  16. Puertas S, Batalla P, Moros M, Polo E, Del Pino P, Guisan JM, Grazu V, de la Fuente JM (2011) Taking advantage of unspecific interactions to produce highly active magnetic nanop-antibody conjugates. ACS Nano 5:4521-4528. https://doi.org/10.1021/nn200019s
  17. Ciminiello P, Dell'Aversano C, Lacovo ED, Fattorusso E, Forino M, Tartaglione L (2011) LC-MS of palytoxin and its analogues: state of the art and future perspectives. Toxicon 57:376-389. https://doi.org/10.1016/j.toxicon.2010.11.002
  18. Luigi Lucini L, Molinari GP (2011) Performance and matrix effect observed in QuEChERS extraction and tandem mass spectrometry analyses of pesticide residues in different target crops. J Chromatogr Sci 49:709-714. https://doi.org/10.1093/chrsci/49.9.709