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Sensitive determination of mixtures of neonicotinoid and fungicide residues in pollen and single bumblebees using a scaled down QuEChERS method for exposure assessment

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Abstract

To accurately estimate exposure of bees to pesticides, analytical methods are needed to enable quantification of nanogram/gram (ng/g) levels of contaminants in small samples of pollen or the individual insects. A modified QuEChERS extraction method coupled with ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analysis was tested to quantify residues of 19 commonly used neonicotinoids and fungicides and the synergist, piperonyl butoxide, in 100 mg samples of pollen and in samples of individual bumblebees (Bombus terrestris). Final recoveries ranged from 71 to 102 % for most compounds with a repeatability of below 20 % for both pollen and bumblebee extracts spiked at 5 and 40 ng/g. The method enables the detection of all compounds at sub-ng/g levels in both matrices and the method detection limits (MDL) ranged from 0.01 to 0.84 ng/g in pollen and 0.01 to 0.96 ng/g in individual bumblebees. Using this method, mixtures of neonicotinoids (thiamethoxam, clothianidin, imidacloprid and thiacloprid) and fungicides (carbendazim, spiroxamine, boscalid, tebuconazole, prochloraz, metconazole, fluoxastrobin, pyraclostrobin and trifloxystrobin) were detected in pollens of field bean, strawberry and raspberry at concentrations ranging from <MDL to 67 ng/g for neonicotinoids and from <MDL to 14 ng/g for fungicides. In bumblebees, the insecticides thiamethoxam and thiacloprid were present at concentrations >MDL, and in some bees, the fungicides carbendazim, boscalid, tebuconazole, flusilazole and metconazole were present at concentrations between 0.80 to 30 ng/g. This new method allows the analysis of mixtures of neonicotinoids and fungicides at trace levels in small quantities of pollen and individual bumblebees and thus will facilitate exposure assessment studies.

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References

  1. Goulson D, Lye GC, Darvill B (2008) Decline and conservation of bumble bees. Annu Rev Entomol 53:191–208

    Article  CAS  Google Scholar 

  2. Goulson D, Nicholls E, Botias C, Rotheray EL (2015) Bee declines driven by combined stress from parasites, pesticides, and lack of flowers. Science 347(6229):122957-1–1255957-9

  3. Rundlof M, Andersson GK, Bommarco R, Fries I, Hederstrom V, Herbertsson L, Jonsson O, Klatt BK, Pedersen TR, Yourstone J, Smith HG (2015) Seed coating with a neonicotinoid insecticide negatively affects wild bees. Nature 521(7550):77–80

    Article  Google Scholar 

  4. Goulson D (2013) REVIEW: an overview of the environmental risks posed by neonicotinoid insecticides. J Appl Ecol 50(4):977–987

    Article  Google Scholar 

  5. Fairbrother A, Purdy J, Anderson T, Fell R (2014) Risks of neonicotinoid insecticides to honeybees. Environ Toxicol Chem 33(4):719–731

    Article  CAS  Google Scholar 

  6. Sanchez-Bayo F, Goka K (2014) Pesticide residues and bees—a risk assessment. PLoS One 9 (4):e94482

  7. Johnson RM (2015) Honey bee toxicology. Annu Rev Entomol 60(60):415–434

    Article  CAS  Google Scholar 

  8. Schmuck R, Stadler T, Schmidt HW (2003) Field relevance of a synergistic effect observed in the laboratory between an EBI fungicide and a chloronicotinyl insecticide in the honeybee (Apis mellifera L, Hymenoptera). Pest Manag Sci 59(3):279–286

    Article  CAS  Google Scholar 

  9. Iwasa T, Motoyama N, Ambrose JT, Roe RM (2004) Mechanism for the differential toxicity of neonicotinoid insecticides in the honey bee, Apis mellifera. Crop Prot 23(5):371–378

    Article  CAS  Google Scholar 

  10. Kosior A, Celary W, Olejniczak P, Fijal J, Krol W, Solarz W, Plonka P (2007) The decline of the bumble bees and cuckoo bees (Hymenoptera: Apidae: Bombini) of Western and Central Europe. Oryx 41(1):79–88

    Article  Google Scholar 

  11. Kamel A (2010) Refined methodology for the determination of neonicotinoid pesticides and their metabolites in honey bees and bee products by liquid chromatography-tandem mass spectrometry (LC-MS/MS). J Agric Food Chem 58(10):5926–5931

    Article  CAS  Google Scholar 

  12. Botías C, David A, Horwood J, Abdul-Sada A, Nicholls E, Hill E, Goulson D (2015) High prevalence of neonicotinoid residues in wildflowers, a potential route of chronic exposure for bees. Environ Sci Technol (submitted)

  13. Anastassiades M, Lehotay SJ, Stajnbaher D, Schenck FJ (2003) Fast and easy multiresidue method employing acetonitrile extraction/partitioning and “dispersive solid-phase extraction” for the determination of pesticide residues in produce. J AOAC Int 86(2):412–431

    CAS  Google Scholar 

  14. Lehotay SJ, Son KA, Kwon H, Koesukwiwat U, Fu W, Mastovska K, Hoh E, Leepipatpiboon N (2010) Comparison of QuEChERS sample preparation methods for the analysis of pesticide residues in fruits and vegetables. J Chromatogr A 1217(16):2548–2560

    Article  CAS  Google Scholar 

  15. Chen M, Collins EM, Tao L, Lu CS (2013) Simultaneous determination of residues in pollen and high-fructose corn syrup from eight neonicotinoid insecticides by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 405(28):9251–9264

    Article  CAS  Google Scholar 

  16. Khan ZS, Ghosh RK, Girame R, Utture SC, Gadgil M, Banerjee K, Reddy DD, Johnson N (2014) Optimization of a sample preparation method for multiresidue analysis of pesticides in tobacco by single and multi-dimensional gas chromatography–mass spectrometry. J Chromatogr A 1343:200–206

    Article  CAS  Google Scholar 

  17. Tascone O, Shirshikova M, Roy C, Meierhenrich UJ (2014) Pesticide determination in rose petals using dispersive solid-phase extraction followed by gas chromatography-tandem mass spectrometry. Anal Bioanal Chem 406(30):8041–8048

    Article  CAS  Google Scholar 

  18. Kasiotis KM, Anagnostopoulos C, Anastasiadou P, Machera K (2014) Pesticide residues in honeybees, honey and bee pollen by LC-MS/MS screening: reported death incidents in honeybees. Sci Total Environ 485–486:633–642

    Article  Google Scholar 

  19. Mullin CA, Frazier M, Frazier JL, Ashcraft S, Simonds R, Vanengelsdorp D, Pettis JS (2010) High levels of miticides and agrochemicals in North American apiaries: implications for honey bee health. PLoS One 5(3), e9754

    Article  Google Scholar 

  20. Wiest L, Bulete A, Giroud B, Fratta C, Amic S, Lambert O, Pouliquen H, Arnaudguilhem C (2011) Multi-residue analysis of 80 environmental contaminants in honeys, honeybees and pollens by one extraction procedure followed by liquid and gas chromatography coupled with mass spectrometric detection. J Chromatogr A 1218(34):5743–5756

    Article  CAS  Google Scholar 

  21. Gbylik-Sikorska M, Sniegocki T, Posyniak A (2015) Determination of neonicotinoid insecticides and their metabolites in honey bee and honey by liquid chromatography tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 990:132–140

    Article  CAS  Google Scholar 

  22. Bingham G, Gunning RV, Delogu G, Borzatta V, Field LM, Moores GD (2008) Temporal synergism can enhance carbamate and neonicotinoid insecticidal activity against resistant crop pests. Pest Manag Sci 64(1):81–85

    Article  CAS  Google Scholar 

  23. Khan HAA, Akram W, Iqbal J, Naeem-Ullah U (2015) Thiamethoxam resistance in the house fly, Musca domestica L.: current status, resistance selection, cross-resistance potential and possible biochemical mechanisms. PLoS One 10 (5):e0125850

  24. Zheng HB, Zhao Q, Mo JZ, Huang YQ, Luo YB, Yu QW, Feng YQ (2013) Quick, easy, cheap, effective, rugged and safe method with magnetic graphitized carbon black and primary secondary amine as adsorbent and its application in pesticide residue analysis. J Chromatogr A 1300:127–133

    Article  CAS  Google Scholar 

  25. Walorczyk S, Gnusowski B (2009) Development and validation of a multi-residue method for the determination of pesticides in honeybees using acetonitrile-based extraction and gas chromatography-tandem quadrupole mass spectrometry. J Chromatogr A 1216(37):6522–6531

    Article  CAS  Google Scholar 

  26. EU (2013) Guidance document on analytical quality control and validation procedures for pesticide residues analysis in food and feed SANCO/12571/2013 rev. 0

  27. Bueno MJM, Boillot C, Munaron D, Fenet H, Casellas C, Gomez E (2014) Occurrence of venlafaxine residues and its metabolites in marine mussels at trace levels: development of analytical method and a monitoring program. Anal Bioanal Chem 406(2):601–610

    Article  Google Scholar 

  28. Walorczyk S (2014) Validation and use of a QuEChERS-based gas chromatographic-tandem mass spectrometric method for multiresidue pesticide analysis in blackcurrants including studies of matrix effects and estimation of measurement uncertainty. Talanta 120:106–113

    Article  CAS  Google Scholar 

  29. Guan YQ, Tang H, Chen DZ, Xu T, Li L (2013) Modified QuEChERS method for the analysis of 11 pesticide residues in tea by liquid chromatography-tandem mass spectrometry. Anal Methods UK 5(12):3056–3067

    Article  CAS  Google Scholar 

  30. Polgar L, Kmellar B, Garcia-Reyes JF, Fodor P (2012) Comprehensive evaluation of the clean-up step in QuEChERS procedure for the multi-residue determination of pesticides in different vegetable oils using LC-MS/MS. Anal Methods UK 4(4):1142–1148

    Article  CAS  Google Scholar 

  31. Christia C, Bizani E, Christophoridis C, Fytianos K (2015) Pesticide residues in fruit samples: comparison of different QuEChERS methods using liquid chromatography-tandem mass spectrometry. Environ Sci Pollut Res Int 22(17):13167–13178

  32. Wong JW, Zhang K, Tech K, Hayward DG, Krynitsky AJ, Cassias I, Schenck FJ, Banerjee K, Dasgupta S, Brown D (2010) Multiresidue pesticide analysis of ginseng powders using acetonitrile- or acetone-based extraction, solid-phase extraction cleanup, and gas chromatography–mass spectrometry/selective ion monitoring (GC-MS/SIM) or -tandem mass spectrometry (GC-MS/MS). J Agr Food Chem 58(10):5884–5896

    Article  CAS  Google Scholar 

  33. Flllion J, Sauve F, Selwyn J (2000) Multiresidue method for the determination of residues of 251 pesticides in fruits and vegetables by gas chromatography/mass spectrometry and liquid chromatography with fluorescence detection. Journal of Aoac International 83(3):698–713

    Google Scholar 

  34. EU (2008) Guidance document on pesticide residue analytical methods. Directorate General Health and Consumer Protection EU guidance SANCO/825/00 rev SANCO/825/00 rev. 8.1

  35. Osborne JL, Clark SJ, Morris RJ, Williams IH, Riley JR, Smith AD, Reynolds DR, Edwards AS (1999) A landscape-scale study of bumble bee foraging range and constancy, using harmonic radar. J Appl Ecol 36(4):519–533

    Article  Google Scholar 

  36. Velthuis HHW, van Doorn A (2006) A century of advances in bumblebee domestication and the economic and environmental aspects of its commercialization for pollination. Apidologie 37(4):421–451

    Article  Google Scholar 

  37. Lye GC, Jennings SN, Osborne JL, Goulson D (2011) Impacts of the use of nonnative commercial bumble bees for pollinator supplementation in raspberry. J Econ Entomol 104(1):107–114

    Article  CAS  Google Scholar 

  38. Foulis ESJ, Goulson D (2014) Commercial bumble bees on soft fruit farms collect pollen mainly from wildflowers rather than the target crops. J Apicult Res 53(3):404–407

    Article  Google Scholar 

  39. Lambert O, Piroux M, Puyo S, Thorin C, L’Hostis M, Wiest L, Bulete A, Delbac F, Pouliquen H (2013) Widespread occurrence of chemical residues in beehive matrices from apiaries located in different landscapes of Western France. PLoS One 8(6), e67007

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful to the Soil Association (Bristol, UK) for part funding of this work.

Conflict of interest

The authors declare that they have no conflict of interest.

Statement on animal ethical care

The work reported here conforms to the regulatory requirements for animal experimentation in the UK. No ethics approval was required for this study. Bumblebee nests were housed on private land for which research permission was granted by the owners. This study did not involve endangered or protected species.

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Authors and Affiliations

  1. School of Life Sciences, University of Sussex, Brighton, Sussex, BN1 9QG, UK

    Arthur David, Cristina Botías, Alaa Abdul-Sada, Dave Goulson & Elizabeth M. Hill

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  1. Arthur David
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  2. Cristina Botías
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  3. Alaa Abdul-Sada
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  4. Dave Goulson
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  5. Elizabeth M. Hill
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Correspondence to Arthur David.

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David, A., Botías, C., Abdul-Sada, A. et al. Sensitive determination of mixtures of neonicotinoid and fungicide residues in pollen and single bumblebees using a scaled down QuEChERS method for exposure assessment. Anal Bioanal Chem 407, 8151–8162 (2015). https://doi.org/10.1007/s00216-015-8986-6

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  • DOI: https://doi.org/10.1007/s00216-015-8986-6

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