Skip to main content

Advertisement

Springer Nature Link
Log in

Untargeted profiling of pesticide metabolites by LC–HRMS: an exposomics tool for human exposure evaluation

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

Abstract

Human exposure to xenobiotics is usually estimated by indirect methods. Biological monitoring has emerged during the last decade to improve assessment of exposure. However, biomonitoring is still an analytical challenge, because the amounts of sample available are often very small yet analysis must be as thorough and sensitive as possible. The purpose of this work was to develop an untargeted “exposomics” approach by using ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC–HRMS), which was applied to the characterization of pesticide metabolites in urine from pregnant women from a French epidemiological cohort. An upgradable list of pesticides commonly used on different crops, with their metabolites (more than 400 substances) was produced. Raw MS data were then processed to extract signals from these substances. Metabolites were identified by tandem mass spectrometry; putative identifications were validated by comparison with standards and metabolites generated by experiments on animals. Finally, signals of identified compounds were statistically analyzed by use of multivariate methods. This enabled discrimination of exposure groups, defined by indirect methods, on the basis of four metabolites from two fungicides (azoxystrobin, fenpropimorph) used in cereal production. This original approach applied to pesticide exposure can be extended to a variety of contaminant families for upstream evaluation of exposure from food and the environment.

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

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

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
Fig. 4
Fig. 5

Similar content being viewed by others

Explore related subjects

Discover the latest articles, books and news in related subjects, suggested using machine learning.

References

  1. US-EPA (2011) Pesticides industry sales and usage. http://www.epa.gov/opp00001/pestsales/07pestsales/market_estimates2007.pdf. 2013

  2. UIPP (2011-2012) Rapport d’activité (Acitivity report). http://asp.zone-secure.net/v2/indexPop.jsp?id=4419/5741/25126&lng=fr. 2013

  3. Mnif W, Hassine AIH, Bouaziz A, Bartegi A, Thomas O, Roig B (2011) Effect of endocrine disruptor pesticides: a review. Int J Environ Res Public Health 8:2265–2303

    Article  CAS  Google Scholar 

  4. Kamel F, Enge LS, Gladen BC, Hoppin JA, Alavanja MCR, Sandler DP (2007) Neurologic symptoms in licensed pesticide applicators in the Agricultural Health Study. Hum Exp Toxicol 26:243–250

    Article  CAS  Google Scholar 

  5. Kamel F, Tanner CM, Umbach DM, Hoppin JA, Alavanja MCR, Blair A, Comyns K, Goldman SM, Korell M, Langston JW, Ross GW, Sandler DP (2007) Pesticide exposure and self-reported Parkinson’s disease in the agricultural health study. Am J Epidemiol 165:364–374

    Article  CAS  Google Scholar 

  6. Blanc-Lapierre A, Bouvier G, Garrigou A, Canal-Raffin M, Raherison C, Brochard P, Baldi I (2012) Effets chroniques des pesticides sur le système nerveux central: état des connaissances épidémiologiques (Chronic effects of pesticides on the central nervous system: current epidemiological knowledge). Rev Epidemiol Santé Publique 60:389–400

    Article  CAS  Google Scholar 

  7. Sever LE, Arbuckle TE, Sweeney A (1997) Reproductive and developmental effects of occupational pesticide exposure: the epidemiologic evidence. Occup Med State Art Rev 12:305–325

    CAS  Google Scholar 

  8. Bretveld RW, Thomas CMG, Scheepers PTJ, Zielhuis GA, Roeleveld N (2006) Pesticide exposure: the hormonal function of the female reproductive system disrupted? Reprod Biol Endocrinol 4:1–14

    Article  Google Scholar 

  9. Bretveld R, Brouwers M, Ebisch I, Roeleveld N (2007) Influence of pesticides on male fertility. Scand J Work Environ Health 33:13–28

    Article  CAS  Google Scholar 

  10. Weselak M, Arbuckle TE, Foster W (2007) Pesticide exposures and developmental outcomes: the epidemiological evidence. J Toxicol Environ Health B Crit Rev 10:41–80

    Article  CAS  Google Scholar 

  11. Dich J, Zahm SH, Hanberg A, Adami HO (1997) Pesticides and cancer. Cancer Causes Control 8:420–443

    Article  CAS  Google Scholar 

  12. Zahm SH, Ward MH, Blair A (1997) Pesticides and cancer. Occup Med State Art Rev 12:269–289

    CAS  Google Scholar 

  13. Merhi M, Raynal H, Cahuzac E, Vinson F, Cravedi JP, Gamet-Payrastre L (2007) Occupational exposure to pesticides and risk of hematopoietic cancers: meta-analysis of case-control studies. Cancer Causes Control 18:1209–1226

    Article  CAS  Google Scholar 

  14. Weichenthal S, Moase C, Chan P (2010) A review of pesticide exposure and cancer incidence in the Agricultural Health Study Cohort. Environ Health Perspect 118:1117–1125

    Article  CAS  Google Scholar 

  15. ANSES (2010) Exposition de la population générale aux résidus de pesticides en France : Synthèse et recommandations du comité d’orientation et de prospective scientifique de l’observatoire des résidus de pesticides, ORP (Exposure of the general population to pesticide residues in France: Summary and recommendations of the steering and the scientific prospective committee of the pesticide residues observatory , PRO). Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail. http://www.observatoire-pesticides.gouv.fr/upload/bibliotheque/171959218396043870616875052847/exposition_population_generale_pesticides_2010_vdef.pdf. 2013

  16. Aprea MC (2012) Environmental and biological monitoring in the estimation of absorbed doses of pesticides. Toxicol Lett 210:110–118

    Article  CAS  Google Scholar 

  17. Needham LL, Calafat AM, Barr DB (2007) Uses and issues of biomonitoring. Int J Hyg Environ Health 210:229–238

    Article  CAS  Google Scholar 

  18. Lu CS, Rodriguez T, Funez A, Irish RS, Fenske RA (2006) The assessment of occupational exposure to diazinon in Nicaraguan plantation workers using saliva biomonitoring. In: Mehlman MA, Soffritti M, Landrigan P, Bingham E, Belpoggi F (ed) Living in a chemical world: framing the future in light of the past, vol 1076. Annals of the New York Academy of Sciences. Blackwell Publishing, Oxford, pp 355-365

  19. Porta M, Puigdomenech E, Ballester F, Selva J, Ribas-Fito N, Llop S, Lopez T (2008) Monitoring concentrations of persistent organic pollutants in the general population: the international experience. Environ Int 34:546–561

    Article  CAS  Google Scholar 

  20. Esteban M, Castano A (2009) Non-invasive matrices in human biomonitoring: a review. Environ Int 35:438–449

    Article  CAS  Google Scholar 

  21. Harris SA, Villeneuve PJ, Crawley CD, Mays JE, Yeary RA, Hurto KA, Meeker JD (2010) National study of exposure to pesticides among professional applicators: an investigation based on urinary biomarkers. J Agric Food Chem 58:10253–10261

    Article  CAS  Google Scholar 

  22. Fujii Y, Ito Y, Harada KH, Hitomi T, Koizumi A, Haraguchi K (2012) Comparative survey of levels of chlorinated cyclodiene pesticides in breast milk from some cities of China, Korea and Japan. Chemosphere 89:452–457

    Article  CAS  Google Scholar 

  23. Bradman A, Whyatt RM (2005) Characterizing exposures to nonpersistent pesticides during pregnancy and early childhood in the National Children’s Study: a review of monitoring and measurement methodologies. Environ Health Perspect 113:1092–1099

    Article  CAS  Google Scholar 

  24. Ueyama J, Saito I, Kamijima M (2010) Analysis and evaluation of pyrethroid exposure in human population based on biological monitoring of urinary pyrethroid metabolites. J Pestic Sci 35:87–98

    Article  CAS  Google Scholar 

  25. Sleeuwenhoek A, Cocker J, Jones K, Cherrie JW (2007) Biological monitoring of pesticide exposures. Research Report TM/07/02. IOM. http://www.iom-world.org/research/report_list.php?pageNum_listReports=1&totalRows_listReports=469. 2011

  26. Bouvier G, Seta N, Vigouroux-Villard A, Blanchard O, Momas I (2005) Insecticide urinary metabolites in nonoccupationally exposed populations. J Toxicol Environ Health B Crit Rev 8:485–512

    Article  CAS  Google Scholar 

  27. Aprea C, Colosio C, Mammone T, Minoia C, Maroni M (2002) Biological monitoring of pesticide exposure: a review of analytical methods. J Chromatogr B 769:191–219

    Article  CAS  Google Scholar 

  28. Barr DB, Needham LL (2002) Analytical methods for biological monitoring of exposure to pesticides: a review. J Chromatogr B 778:5–29

    Article  CAS  Google Scholar 

  29. Margariti MG, Tsakalof AK, Tsatsakis AM (2007) Analytical methods of biological monitoring for exposure to pesticides: recent update. Ther Drug Monit 29:150–163

    Article  CAS  Google Scholar 

  30. Le Bizec B, Pinel G, Antignac JP (2009) Options for veterinary drug analysis using mass spectrometry. J Chromatogr A 1216:8016–8034

    Article  Google Scholar 

  31. Bicker W, Lammerhofer M, Genser D, Kiss H, Lindner W (2005) A case study of acute human chlorpyrifos poisoning: novel aspects on metabolism and toxicokinetics derived from liquid chromatography-tandem mass spectrometry analysis of urine samples. Toxicol Lett 159:235–251

    Article  CAS  Google Scholar 

  32. Ahn KC, Watanabe T, Gee SJ, Hammock BD (2004) Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin. J Agric Food Chem 52:4583–4594

    Article  CAS  Google Scholar 

  33. Shim WB, Yakovleva ME, Kim KY, Nam BR, Vylegzhanina ES, Komarov AA, Eremin SA, Chung DH (2009) Development of fluorescence polarization immunoassay for the rapid detection of 6-chloronicotinic acid: main metabolite of neonicotinoid insecticides. J Agric Food Chem 57:791–796

    Article  CAS  Google Scholar 

  34. Colume A, Cardenas S, Gallego M, Valcarcel M (2001) A solid phase extraction method for the screening and determination of pyrethroid metabolites and organochlorine pesticides in human urine. Rapid Commun Mass Spectrom 15:2007–2013

    Article  CAS  Google Scholar 

  35. Maurer HH (2007) Current role of liquid chromatography-mass spectrometry in clinical and forensic toxicology. Anal Bioanal Chem 388:1315–1325

    Article  CAS  Google Scholar 

  36. Dresen S, Ferreiros N, Gnann H, Zimmermann R, Weinmann W (2010) Detection and identification of 700 drugs by multi-target screening with a 3200 Q TRAP(A (R)) LC-MS–MS system and library searching. Anal Bioanal Chem 396:2425–2434

    Article  CAS  Google Scholar 

  37. Polettini A, Gottardo R, Pascali JP, Tagliaro F (2008) Implementation and performance evaluation of a database of chemical formulas for the screening of pharmaco/toxicologically relevant compounds in biological samples using electrospray ionization-time-of-flight mass spectrometry. Anal Chem 80:3050–3057

    Article  CAS  Google Scholar 

  38. Pelander A, Ojanpera I, Laks S, Rasanen I, Vuori E (2003) Toxicological screening with formula-based metabolite identification by liquid chromatography/time-of-flight mass spectrometry. Anal Chem 75:5710–5718

    Article  CAS  Google Scholar 

  39. Lee HK, Ho CS, Iu YPH, Lai PSJ, Shek CC, Lo YC, Klinke HB, Wood M (2009) Development of a broad toxicological screening technique for urine using ultra-performance liquid chromatography and time-of-flight mass spectrometry. Anal Chim Acta 649:80–90

    Article  CAS  Google Scholar 

  40. Broecker S, Herre S, Wust B, Zweigenbaum J, Pragst F (2011) Development and practical application of a library of CID accurate mass spectra of more than 2,500 toxic compounds for systematic toxicological analysis by LC-QTOF-MS with data-dependent acquisition. Anal Bioanal Chem 400:101–117

    Article  CAS  Google Scholar 

  41. Peters RJB, Stolker AAM, Mol JGJ, Lommen A, Lyris E, Angelis Y, Vonaparti A, Stamou M, Georgakopoulos C, Nielen MWF (2010) Screening in veterinary drug analysis and sports doping control based on full-scan, accurate-mass spectrometry. Trends Anal Chem 29:1250–1268

    Article  CAS  Google Scholar 

  42. Petit C, Blangiardo M, Richardson S, Coquet F, Chevrier C, Cordier S (2002) Association of environmental insecticide exposure and fetal growth with a Bayesian model including multiple exposure sources. Am J Epidemiol 175:1182–1190

    Article  Google Scholar 

  43. Agreste (2000) Recensement agricole (Agricultural census). Ministere de l’agriculture, de l’agroalimentaire et de la forêt. http://agreste.agriculture.gouv.fr/enquetes/recensement-agricole/. 2011

  44. Wold S, Antti H, Lindgren F, Ohman J (1998) Orthogonal signal correction of near-infrared spectra. Chemometr Intell Lab Syst 44:175–185

    Article  CAS  Google Scholar 

  45. Dettmer K, Aronov PA, Hammock BD (2007) Mass spectrometry-based metabolomics. Mass Spectrom Rev 26:51–78

    Article  CAS  Google Scholar 

  46. COREP Bretagne (2011) Liste des études suivies par la COREP (List of studies undertaken by COREP). Ministere de l’agriculture, de l’agroalimentaire et de la forêt. http://draf.bretagne.agriculture.gouv.fr/corpep/spip.php?article166. 2011

  47. Association de Coordination Technique Agricole ACTA, Couteux A, Lejeune V (2004) Index phytosanitaire 2004. ACTA, 149 rue de Bercy, Paris, France

  48. IPCS INCHEM (2011) Pesticides data sheets and environmental health criteria. http://www.inchem.org/pages/pds.html. 2011

  49. European Food Safety Authority (2010) 2008 annual report on pesticide residues. EFSA J 8:1646–2088

    Google Scholar 

  50. University of Hertfordshire (2011) PPDB: Pesticide Properties DataBase. http://sitem.herts.ac.uk/aeru/footprint/en/index.htm. 2011

  51. Roberts TR (1999) Metabolic pathways of agrochemicals: part 2: insecticides and fungicides. Royal Society of Chemistry, Cambridge

    Book  Google Scholar 

  52. Roberts TR, Hutson DH (1998) Metabolic pathways of agrochemicals: part 1: herbicides and plant growth regulators. Royal Society of Chemistry, Cambridge

    Book  Google Scholar 

  53. Ueyama J, Saito I, Kondo T, Taki T, Kimata A, Saito S, Ito Y, Murata K, Iwata T, Gotoh M, Shibata E, Wakusawa S, Kamijima M (2012) Urinary concentrations of organophosphorus insecticide metabolites in Japanese workers. Chemosphere 87:1403–1409

    Article  CAS  Google Scholar 

  54. Sumner LW, Amberg A, Barrett D, Beale MH, Beger R, Daykin CA, Fan TWM, Fiehn O, Goodacre R, Griffin JL, Hankemeier T, Hardy N, Harnly J, Higashi R, Kopka J, Lane AN, Lindon JC, Marriott P, Nicholls AW, Reily MD, Thaden JJ, Viant MR (2007) Proposed minimum reporting standards for chemical analysis. Metabolomics 3:211–221

    Article  CAS  Google Scholar 

  55. Krieger RI, Chen L, Ginevan M, Watkins D, Cochran RC, Driver JH, Ross JH (2012) Implications of estimates of residential organophosphate exposure from dialkylphosphates (DAPs) and their relevance to risk. Regul Toxicol Pharmacol 64:263–266

    Article  CAS  Google Scholar 

  56. Hays SM, Aylward LL, Driver J, Ross J, Kirman C (2012) 2,4-D exposure and risk assessment: comparison of external dose and biomonitoring based approaches. Regul Toxicol Pharmacol 64:481–489

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Marion Gaudicheau for her participation in this work. Elisabeth Jeunesse and Florence Blas-y-Estrada (INRA Toxalim, Toulouse, France) are also thanked for the animal experiment. This work was achieved within the MetaToul–AXIOM analytical program and was supported by INRA-AlimH research division and by the French Ministry of Environment in a national program Grenelle de l’Environnement programme LOLF 189 funding (EXPiOmark).

Author information

Authors and Affiliations

  1. INRA, UMR1331, Toxalim, Research Centre in Food Toxicology, 180 Chemin de Tournefeuille, 31027, Toulouse, France

    Emilien L. Jamin, Nathalie Bonvallot, Marie Tremblay-Franco, Jean-Pierre Cravedi & Laurent Debrauwer

  2. Toulouse University, INP, UMR1331, Toxalim, 180 Chemin de Tournefeuille, 31027, Toulouse, France

    Emilien L. Jamin, Nathalie Bonvallot, Marie Tremblay-Franco, Jean-Pierre Cravedi & Laurent Debrauwer

  3. INSERM UMR 1085 IRSET – Institut de Recherche en Santé, Environnement, Travail, Avenue du Général Leclerc, 35042, Rennes, France

    Nathalie Bonvallot, Cécile Chevrier & Sylvaine Cordier

  4. EHESP – School of Public Health, Avenue du Général Leclerc, 35042, Rennes - Sorbonne Paris Cité, France

    Nathalie Bonvallot

  5. University of Rennes I, Avenue du Général Leclerc, 35042, Rennes, France

    Cécile Chevrier & Sylvaine Cordier

Authors
  1. Emilien L. Jamin
    View author publications

    Search author on:PubMed Google Scholar

  2. Nathalie Bonvallot
    View author publications

    Search author on:PubMed Google Scholar

  3. Marie Tremblay-Franco
    View author publications

    Search author on:PubMed Google Scholar

  4. Jean-Pierre Cravedi
    View author publications

    Search author on:PubMed Google Scholar

  5. Cécile Chevrier
    View author publications

    Search author on:PubMed Google Scholar

  6. Sylvaine Cordier
    View author publications

    Search author on:PubMed Google Scholar

  7. Laurent Debrauwer
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Emilien L. Jamin.

Additional information

Published in the special issue Analytical Science in France with guest editors Christian Rolando and Philippe Garrigues.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jamin, E.L., Bonvallot, N., Tremblay-Franco, M. et al. Untargeted profiling of pesticide metabolites by LC–HRMS: an exposomics tool for human exposure evaluation. Anal Bioanal Chem 406, 1149–1161 (2014). https://doi.org/10.1007/s00216-013-7136-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-013-7136-2

Keywords