Skip to main content
SpringerLink
Log in

Determination of highly polar anionic pesticides in beehive products by hydrophilic interaction liquid chromatography coupled to mass spectrometry

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

The analysis of highly polar pesticides is challenging due to their unique physicochemical properties, requiring specialized chromatographic techniques for their accurate and sensitive detection. Furthermore, the high level of co-extracted polar matrix components that can co-elute with the analytes can interfere with the analysis. Consequently, there is lack of pesticide monitoring data, as the European Food Safety Authority has pointed out. This article explores the overcoming of such difficulties in the analysis of these compounds. Analytical methodologies for the extraction, clean-up, and direct determination of 11 highly polar anionic pesticides, including glyphosate, glufosinate, ethephon, fosetyl-aluminium, and their related metabolites in complex food matrices such as honey and pollen by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry were successfully developed and validated. Solid-phase extraction and micro-solid-phase extraction employing strong anion exchange (SAX) cartridges were implemented for clean-up. The automation and miniaturization of SAX clean-up for these compounds were achieved for the first time. For method validation, SANTE/11312/2021 guideline was followed. Recoveries were between 70 and 120%, with RSDs below 20%. Limits of quantitation ranged from 0.005 to 0.020 mg kg-1. Linearity was evaluated from 0.002 to 0.200 mg kg-1. Matrix effects were assessed, showing medium to low signal suppression for most compounds. AMPA and glufosinate presented the highest signal suppression, but it was reduced after SAX clean-up. Analysis of real honey and pollen samples revealed the occurrence of the studied compounds in beehive products and showed the applicability of the validated methodologies for routine control of these complex samples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. EFSA, Carrasco Cabrera L, Medina Pastor P. The 2020 European Union report on pesticide residues in food. EFSA J. 2022;20(3):e07215. https://doi.org/10.2903/j.efsa.2022.7215.

    Article  Google Scholar 

  2. Melton LM, Taylor MJ, Flynn EE. The utilisation of ion chromatography and tandem mass spectrometry (IC-MS/MS) for the multi-residue simultaneous determination of highly polar anionic pesticides in fruit and vegetables. Food Chem. 2019;298:125028. https://doi.org/10.1016/j.foodchem.2019.125028.

    Article  CAS  PubMed  Google Scholar 

  3. Guo Y, Gaiki S. Retention and selectivity of stationary phases for hydrophilic interaction chromatography. J Chromatogr A. 2011;1218(35):5920–38. https://doi.org/10.1016/j.chroma.2011.06.052.

    Article  CAS  PubMed  Google Scholar 

  4. Botero-Coy AM, Ibáñez M, Sancho JV, Hernández F. Direct liquid chromatography–tandem mass spectrometry determination of underivatized glyphosate in rice, maize and soybean. J Chromatogr A. 2013;1313:157–65. https://doi.org/10.1016/j.chroma.2013.07.037.

    Article  CAS  PubMed  Google Scholar 

  5. Cutillas V, Fernández-Alba AR. Analysis by LC-MS/MS of polar pesticides in fruits and vegetables using new hybrid stationary phase. MethodsX. 2021;8:101306. https://doi.org/10.1016/j.mex.2021.101306.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Herrera López S, Scholten J, Kiedrowska B, de Kok A. Method validation and application of a selective multiresidue analysis of highly polar pesticides in food matrices using hydrophilic interaction liquid chromatography and mass spectrometry. J Chromatogr A. 2019;1594:93–104. https://doi.org/10.1016/j.chroma.2019.02.024.

    Article  CAS  PubMed  Google Scholar 

  7. Herrera López S, Dias J, de Kok A. Analysis of highly polar pesticides and their main metabolites in animal origin matrices by hydrophilic interaction liquid chromatography and mass spectrometry. Food Control. 2020;115:107289. https://doi.org/10.1016/j.foodcont.2020.107289.

    Article  CAS  Google Scholar 

  8. Herrera Lopez S, Dias J, Mol H, de Kok A. Selective multiresidue determination of highly polar anionic pesticides in plant-based milk, wine and beer using hydrophilic interaction liquid chromatography combined with tandem mass spectrometry. J Chromatogr A. 2020;1625:461226. https://doi.org/10.1016/j.chroma.2020.461226.

    Article  CAS  Google Scholar 

  9. Kaczyński P. Clean-up and matrix effect in LC-MS/MS analysis of food of plant origin for high polar herbicides. Food Chem. 2017;230:524–31. https://doi.org/10.1016/j.foodchem.2017.03.091.

    Article  CAS  PubMed  Google Scholar 

  10. Wu Y, et al. Development and inter-laboratory validation of analytical methods for glufosinate and its two metabolites in foods of plant origin. Anal Bioanal Chem. 2023. https://doi.org/10.1007/s00216-023-04542-9.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Chamkasem N, Morris C, Harmon T. Direct determination of glyphosate, glufosinate, and AMPA in milk by liquid chromatography/tandem mass spectrometry. J Regul Sci. 2015;02:20–6.

    Google Scholar 

  12. Hakme E, Poulsen ME. Evaluation of the automated micro-solid phase extraction clean-up system for the analysis of pesticide residues in cereals by gas chromatography-Orbitrap mass spectrometry. J Chromatogr A. 2021;1652:462384. https://doi.org/10.1016/j.chroma.2021.462384.

    Article  CAS  PubMed  Google Scholar 

  13. Nagatomi Y, Yoshioka T, Yanagisawa M, Uyama A, Mochizuki N. Simultaneous LC-MS/MS analysis of glyphosate, glufosinate, and their metabolic products in beer, barley tea, and their ingredients. Biosci Biotechnol Biochem. 2013;77(11):2218–21. https://doi.org/10.1271/bbb.130433.

    Article  CAS  PubMed  Google Scholar 

  14. Ding J, et al. Determination of underivatized glyphosate residues in plant-derived food with low matrix effect by solid phase extraction-liquid chromatography-tandem mass spectrometry. Food Anal Methods. 2016;9(10):2856–63. https://doi.org/10.1007/s12161-016-0468-8.

    Article  Google Scholar 

  15. Gotti R, Fiori J, Bosi S, Dinelli G. Field-amplified sample injection and sweeping micellar electrokinetic chromatography in analysis of glyphosate and aminomethylphosphonic acid in wheat. J Chromatogr A. 2019;1601:357–64. https://doi.org/10.1016/j.chroma.2019.05.013.

    Article  CAS  PubMed  Google Scholar 

  16. Lehotay SJ, Han L, Sapozhnikova Y. Automated mini-column solid-phase extraction cleanup for high-throughput analysis of chemical contaminants in foods by low-pressure gas chromatography—tandem mass spectrometry. Chromatographia. 2016;79(17):1113–30. https://doi.org/10.1007/s10337-016-3116-y.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Michlig N, Lehotay SJ. Evaluation of a septumless mini-cartridge for automated solid-phase extraction cleanup in gas chromatographic analysis of >250 pesticides and environmental contaminants in fatty and nonfatty foods. J Chromatogr A. 2022;1685:463596. https://doi.org/10.1016/j.chroma.2022.463596.

    Article  CAS  PubMed  Google Scholar 

  18. Manzano Sánchez L, Jesús F, Ferrer C, Gómez-Ramos MM, Fernández-Alba A. Evaluation of automated clean-up for large scope pesticide multiresidue analysis by liquid chromatography coupled to mass spectrometry. J Chromatogr A. 2023;1694:463906. https://doi.org/10.1016/j.chroma.2023.463906.

    Article  CAS  PubMed  Google Scholar 

  19. Murcia-Morales M, Heinzen H, Parrilla-Vázquez P, Gómez-Ramos MM, Fernández-Alba AR. Presence and distribution of pesticides in apicultural products: a critical appraisal. TrAC Trends Anal Chem. 2022;146. https://doi.org/10.1016/j.trac.2021.116506.

  20. Rampazzo G, Gazzotti T, Zironi E, Pagliuca G. Glyphosate and glufosinate residues in honey and other hive products. Foods. 2023;12(6). 10.3390/foods12061155.

  21. Verdini E, Pecorelli I. The current status of analytical methods applied to the determination of polar pesticides in food of animal origin: a brief review. Foods. 2022;11(10). 10.3390/foods11101527.

  22. Thompson HM, et al. Evaluating exposure and potential effects on honeybee brood (Apis mellifera) development using glyphosate as an example. Integr Environ Assess Manag. 2014;10(3):463–70. https://doi.org/10.1002/ieam.1529.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Thompson TS, van den Heever JP, Limanowka RE. Determination of glyphosate, AMPA, and glufosinate in honey by online solid-phase extraction-liquid chromatography-tandem mass spectrometry. Food Addit Contam Part A. 2019;36(3):434–46. https://doi.org/10.1080/19440049.2019.1577993.

    Article  CAS  Google Scholar 

  24. El Agrebi N, Tosi S, Wilmart O, Scippo M-L, de Graaf DC, Saegerman C. Honeybee and consumer’s exposure and risk characterisation to glyphosate-based herbicide (GBH) and its degradation product (AMPA): residues in beebread, wax, and honey. Sci Total Environ. 2020;704:135312. https://doi.org/10.1016/j.scitotenv.2019.135312.

    Article  CAS  PubMed  Google Scholar 

  25. Odemer R et al. Chronic high glyphosate exposure delays individual worker bee (Apis mellifera L.) development under field conditions. Insects. 2020;11(10). https://doi.org/10.3390/insects11100664.

  26. Bergero M, Bosco L, Giacomelli A, Angelozzi G, Perugini M, Merola C. Agrochemical contamination of honey and bee bread collected in the Piedmont Region, Italy. Environments. 2021;8(7). https://doi.org/10.3390/environments8070062.

  27. Pareja L, Jesús F, Heinzen H, Hernando MD, Rajski Ł, Fernández-Alba AR. Evaluation of glyphosate and AMPA in honey by water extraction followed by ion chromatography mass spectrometry. A pilot monitoring study. Anal Methods. 2019;11(16):2123–8. https://doi.org/10.1039/C9AY00543A.

    Article  CAS  Google Scholar 

  28. Gasparini M, Angelone B, Ferretti E. Glyphosate and other highly polar pesticides in fruit, vegetables and honey using ion chromatography coupled with high resolution mass spectrometry: method validation and its applicability in an official laboratory. J Mass Spectrom. 2020;55(11):e4624. https://doi.org/10.1002/jms.4624.

    Article  CAS  PubMed  Google Scholar 

  29. Gómez IB, Ramos MJG, Rajski Ł, Flores JM, Jesús F, Fernández-Alba AR. Ion chromatography coupled to Q-Orbitrap for the analysis of formic and oxalic acid in beehive matrices: a field study. Anal Bioanal Chem. 2022;414(7):2419–30. https://doi.org/10.1007/s00216-022-03882-2.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. EFSA. Review of the existing maximum residue levels for glyphosate according to Article 12 of Regulation (EC) No 396/2005 – revised version to take into account omitted data. EFSA J. 2019;17(10):5862. https://doi.org/10.2903/j.efsa.2019.5862.

    Article  Google Scholar 

  31. Dias J, López SH, Mol H, de Kok A. Influence of different hydrophilic interaction liquid chromatography stationary phases on method performance for the determination of highly polar anionic pesticides in complex feed matrices. J Sep Sci. 2021;44(11):2165–76. https://doi.org/10.1002/jssc.202001134.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Takayama N, Lim LW, Takeuchi T. Retention behavior of inorganic anions in hydrophilic interaction chromatography. Anal Sci. 2017;33(5):619–23. https://doi.org/10.2116/analsci.33.619.

    Article  CAS  PubMed  Google Scholar 

  33. Document No. SANTE/11312/2021. Analytical quality control and method validation procedures for pesticide residues analysis in food and feed. https://www.eurl-pesticides.eu/userfiles/file/EurlALL/SANTE_11312_2021.pdf. Accessed 3 Jul 2023.

Download references

Acknowledgements

The authors kindly acknowledge CTC Analytics for providing custom-made SAX µSPE cartridges, and especially Dr. Hans-Joachim Hübschmann for his technical support.

Author information

Authors and Affiliations

  1. Department of Chemistry and Physics, Agrifood Campus of International Excellence (ceiA3), University of Almeria, Ctra. Sacramento s/n, La Cañada de San Urbano, 04120, Almeria, Spain

    Florencia Jesús, Adrián Rosa García, Víctor Cutillas & Amadeo Rodríguez Fernández-Alba

  2. Chemistry Interdisciplinary Project (ChIP), School of Pharmacy, University of Camerino, Via Madonna delle Carceri, 62032, Camerino, Italy

    Tommaso Stecconi

Authors
  1. Florencia Jesús
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Adrián Rosa García
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tommaso Stecconi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Víctor Cutillas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Amadeo Rodríguez Fernández-Alba
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Amadeo Rodríguez Fernández-Alba.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Published in the topical collection Food Safety Analysis 2.0 with guest editor Steven J. Lehotay.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 588 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jesús, F., Rosa García, A., Stecconi, T. et al. Determination of highly polar anionic pesticides in beehive products by hydrophilic interaction liquid chromatography coupled to mass spectrometry. Anal Bioanal Chem 416, 675–688 (2024). https://doi.org/10.1007/s00216-023-04946-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-023-04946-7

Keywords

Search

Navigation