Advertisement

Food Analytical Methods

, Volume 10, Issue 12, pp 4013–4023 | Cite as

High-affinity Antibodies from a Full Penthiopyrad-mimicking Hapten and Heterologous Immunoassay Development for Fruit Juice Analysis

  • Eric Ceballos-Alcantarilla
  • Consuelo Agulló
  • Antonio Abad-Fuentes
  • Mónica Escamilla-Aguilar
  • Antonio Abad-Somovilla
  • Josep V. MercaderEmail author
Article
  • 131 Downloads

Abstract

New-generation succinate dehydrogenase inhibitors with increased antifungal performance have been developed during the last years. Particularly, penthiopyrad has been recently introduced in the agrochemical market as a complementary fungicide for combined treatments with other active principles displaying a different mode of action. In the present study, rapid and high-throughput analytical methods for penthiopyrad residue determination in foodstuffs were developed. A novel functionalized hapten that completely mimics the molecule of penthiopyrad was designed for high-affinity and specific antibody generation. Moreover, heterologous haptens were synthesized, and the influence of their respective molecular modifications over antibody binding was evaluated by competitive enzyme-linked immunosorbent assay. Two immunoassays were optimized using alternative assay formats, affording limits of detection for penthiopyrad in buffer around 0.07 ng/mL in both cases. Performance, in terms of accuracy and precision, of the developed immunochemical methods was studied using apple juice and red and white grape juices. The obtained results showed that penthiopyrad could be determined in those food samples in compliance with European maximum residue limits.

Keywords

Target mimicking Antibodies Fungicide residues ELISA Immunoassay 

Abbreviations

ABS

adult bovine serum

BSA

bovine serum albumin

DMF

N,N′-dimethylformamide

DSC

N,N′-disuccinimidyl carbonate

ELISA

enzyme-linked immunosorbent assay

ESI

electrospray ionization

GAR-HRP

goat antirabbit immunoglobulins conjugated to peroxidase

HRMS

high-resolution mass spectrometry

HRP

horseradish peroxidase

IR

infrared

LOD

limit of detection

MR

molar ratio

MRL

maximum residue limit

OVA

ovalbumin

PB

phosphate buffer

PBS

phosphate-buffered saline

PBST

PBS containing Tween 20

SDHI

succinate dehydrogenase inhibitor

TLC

thin-layer chromatography

UV

ultraviolet

Notes

Acknowledgements

This work was supported by the Spanish Ministerio de Ciencia e Innovación (AGL2012-39965-C02 and AGL2015-64488-C2) and cofinanced by FEDER funds. E.C.-A. is recipient of a predoctoral fellowship from the “Atracció de Talent, VLC-CAMPUS” program of the University of Valencia. Conjugate analysis was carried out at the Proteomics Unit—a member of ISCIII ProteoRedProteomics Platform—and animal manipulation was carried out at the Animal Production Section, both belonging to the SCSIE of the University of Valencia.

Compliance with Ethical Standards

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Conflict of Interest

Eric Ceballos-Alcantarilla declares that he has no conflict of interest. Consuelo Agulló declares that she has no conflict of interest. Antonio Abad-Fuentes declares that he has no conflict of interest. Mónica Escamilla-Aguilar declares that she has no conflict of interest. Antonio Abad-Somovilla declares that he has no conflict of interest. Josep V. Mercader declares that he has no conflict of interest.

References

  1. Abad-Fuentes A, Ceballos-Alcantarilla E, Mercader JV, Agulló C, Abad-Somovilla A, Esteve-Turrillas FA (2015) Determination of succinate-dehydrogenase-inhibitor fungicide residues in fruits and vegetables by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 407:4207–4211CrossRefGoogle Scholar
  2. AgroNews. (2017) Global SDHI fungicide market expected to reach $4 bn in 2020. www.agropages.com. Accessed 22 May 2017
  3. Ahn KC, Kim HJ, McCoy MR, Gee SJ, Hammock BD (2011) Immunoassays and biosensors for monitoring environmental and human exposure to pyrethroid insecticides. J Agric Food Chem 59:2792–2802CrossRefGoogle Scholar
  4. APVMA, Australian Pesticides and Veterinary Medicines Authority (2012) Public release summary on the evaluation of penthiopyrad in the new product DuPont™ Fontelis® fungicide. APVMA Product Number 65100Google Scholar
  5. Armarego WLF, Perrin DD (1997) In: purification of laboratory chemicals, 4th edn. Oxford, Butterworth-Heinemann LtdGoogle Scholar
  6. Avenot HF, Michailides TJ (2010) Progress in understanding molecular mechanisms and evolution of resistance to succinate dehydrogenase inhibiting (SDHI) fungicides in phytopathogenic fungi. Crop Prot 29:643–651CrossRefGoogle Scholar
  7. Ceballos-Alcantarilla E, Abad-Fuentes A, Aloisio V, Agulló C, Abad-Somovilla A, Mercader JV (2014) Haptens, bioconjugates, and antibodies for penthiopyrad immunosensing. Analyst 139:5338–5361CrossRefGoogle Scholar
  8. Culbreath AK, Brenneman TB, Kemerait RC Jr, Hammes GG (2009) Effect of the new pyrazole carboxamide fungicide penthiopyrad on late leaf spot and stem rot of peanut. Pest Manag Sci 65:66–73CrossRefGoogle Scholar
  9. Dankwardt A (2006) Immunochemical assays in pesticide analysis. In: Meyers RA (ed) Encyclopedia of analytical chemistry. John Wiley & Sons Ltd, Chichester, UK, pp 1–27Google Scholar
  10. DG SANCO (2013) Final review report for the active substance penthiopyrad. Document number SANCO/12078/2013 rev 2Google Scholar
  11. EFSA, European Food Safety Authority (2013) Conclusion on the peer review of the pesticide risk assessment of the active substance penthiopyrad. EFSA J 11(2):3111CrossRefGoogle Scholar
  12. Esteve-Turrillas FA, Parra J, Abad-Fuentes A, Agulló C, Abad-Somovilla A, Mercader JV (2010) Hapten synthesis, monoclonal antibody generation, and development of competitive immunoassays for the analysis of picoxystrobin in beer. Anal Chim Acta 682:93–103CrossRefGoogle Scholar
  13. FAO (2011) Penthiopyrad. In: Joint FAO/WHO Meeting on Pesticide Residues. Pesticide Residues in Food 2011. FAO Plant Production and Protection Paper 211:189–194Google Scholar
  14. Gudmestad NC, Arabiat S, Miller JS, Pasche JS (2013) Prevalence and impact of SDHI fungicide resistance in Alternaria solani. Plant Dis 97(7):952–960CrossRefGoogle Scholar
  15. Gulkowska A, Buerge IJ, Poiger T (2014) Online solid phase extraction LC-MS/MS method for the analysis of succinate dehydrogenase inhibitor fungicides and its applicability to surface water samples. Anal Bioanal Chem 406:6419–6427CrossRefGoogle Scholar
  16. Li YF, Sun YM, Beier RC, Lei HT, Gee S, Hammock BD, Wang H, Wang Z, Sun X, Shen YD, Yang JY, Xu ZL (2017) Immunochemical techniques for multianalyte analysis of chemical residues in food and the environment: a review. TRAC-Trend Anal Chem 88:25–40CrossRefGoogle Scholar
  17. López-Moreno R, Mercader JV, Agulló C, Abad-Somovilla A, Abad-Fuentes A (2013) Structure–immunogenicity relationship of kresoxim-methyl regioisomeric haptens. Org Biomol Chem 11:7361–7371CrossRefGoogle Scholar
  18. Pesticide Properties Database. (2017) http://sitem.herts.ac.uk/aeru/ppdb/en/. Accessed 22 May 2017
  19. Proffer TJ, Lizotte E, Rothwell NL, Sundin GW (2013) Evaluation of dodine, fluopyram and penthiopyrad for the management of leaf spot and powdery mildew of tart cherry, and fungicide sensitivity screening of Michigan populations of Blumeriella jaapii. Pest Manag Sci 69:747–754CrossRefGoogle Scholar
  20. Sakurai S, Hagiwara H, Yanase Y (2011) Biological activity of penthiopyrad and study on sensitivity test for several plant pathogens. J Pestic Sci 36(4):520–523CrossRefGoogle Scholar
  21. Suri CR, Boro R, Nangia Y, Gandhi S, Sharma P, Wangoo N, Rajesh K (2009) Immunoanalytical techniques for analyzing pesticides in the environment. TRAC-Trend Anal Chem 28:29–39CrossRefGoogle Scholar
  22. Van Emon JM, Chuang JC, Dill K, Xiong G (2008) Immunoassays and biosensors. In: Tadeo JL (ed) Analysis of pesticides in food and environmental samples. CRC Press, Taylor & Francis Group, Boca Raton, pp 95–123Google Scholar
  23. Xu ZL, Shen YD, Beier RC, Yang JY, Lei HT, Wang H, Sun YM (2009) Application of computer-assisted molecular modeling for immunoassay of low molecular weight food contaminants: a review. Anal Chim Acta 647:125–136CrossRefGoogle Scholar
  24. Yoshikawa Y, Katsuta H, Kishi J, Yanase Y (2011) Structure–activity relationship of carboxin-related carboxamides as fungicide. J Pestic Sci 36(3):347–356CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Department of Organic ChemistryUniversitat de ValènciaBurjassotSpain
  2. 2.Department of Technologies for Food Preservation and SafetyInstitute of Agrochemistry and Food Technology (IATA), Spanish National Research Council (CSIC)PaternaSpain

Personalised recommendations