Development and application of a QuEChERS-based extraction method for the analysis of 55 pesticides in the bivalve Scrobicularia plana by GC-MS/MS
- 492 Downloads
A method for quantitative determination of 55 pesticides in a bivalve matrix was established, based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction and using gas chromatography (GC)-ion trap (IT) mass spectrometry (MS/MS). Accomplishing the European SANCO guidelines, this method was validated using 5 g of homogenized soft tissue, allowing the quantification of pesticides at ng/g of wet weight (ww). Quantification limits and recovery rates ranged from 0.33 to 10.3 μg/L and from 78 to 119 %, respectively. As an important mollusc, not only from an ecological perspective but also for food consumption, the peppery furrow shell (Scrobicularia plana) was sampled at three strategical sites (Ria Formosa Lagoon, in the south of Portugal) during 2012–2013, over six campaigns. A total of 2160 animals were pooled by place and sex. No statistical differences were found among sites or between sexes. Forty percent of the sampled pools were above quantification limits, reaching total annual average concentrations of ∑800 ng/g ww. Additionally, 83 % of the selected compounds showed concentrations above the legal limits set by the European Directive 2013/39/EU. In conclusion, the applied method was successful and proved that bivalves were contaminated by the selected pesticides. In future work, this methodology can be used to monitor body burdens and obtain data for predicting impacts in shellfish consumers.
Keywords2013/39/EU Fungicides Herbicides Insecticides SANCO/825/00 Seafood
This research was partially supported by the European Regional Development Fund (ERDF) through the COMPETE - Operational Competitiveness Programme, and POPH – Operational Human Potential Programme, and by national Portuguese funds, through FCT – Foundation for Science and Technology, via the strategic funding project UID/Multi/04423/2013, project PTDC/MAR/70436/2006 (FCOMP-01-0124.FEDER-7382), and, finally, the PhD grant attributed to C.C. (SFRH/BD/79305/2011).
The authors thank the expert advice and help offered by Célia Lopes to perform the Diff-Quick staining of the bivalves’ gonad squashes. A special thanks is also extended to Sukanlaya Tantiwisawaruji for the help when staining the cited squashes. Acknowledgements are also due to Ana Valente, PhD for proofreading the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
All the animals received human care and all experimental protocols were performed in accordance with the Portuguese Animal Welfare Law (Decreto-Lei no.113/2013, 7 de Agosto D.R. no. 151, Série I) and animal protocols approved by CIIMAR/UP and DGAV (Direcção-Geral de Alimentação e Veterinária, the Portuguese National Authority for Animal Health).
- 1.US Environmental Protection Agency (EPA). Persistent organic pollutants: a global issue, a global response: Office of International Affairs, US EPA; 2002, p. 26Google Scholar
- 4.Badach H, Nazimek T, Kaminska IA. Pesticide content in drinking water samples collected from orchard areas in central Poland. Ann Agric Environ Med. 2007;14(1):109.Google Scholar
- 8.Katagi T. Bioconcentration, bioaccumulation, and metabolism of pesticides in aquatic organisms. In: Whitacre DM, editor. Reviews of Environmental Contamination and Toxicology. New York: Springer; 2010. p. 1–132.Google Scholar
- 13.Tsygankov VY, Boyarova MD, Lukyanova ON. Bioaccumulation of persistent organochlorine pesticides (OCPs) by gray whale and Pacific walrus from the western part of the Bering Sea. Mar Pollut Bull. 2015;99(1-2):235–39.Google Scholar
- 17.Instituto nacional de estatística (INE). Estatística da pesca 2010. In: Direção-geral de recursos naturais segurança e serviços marítimos, pp. 101; INE: Lisbon, PortugalGoogle Scholar
- 18.Food balance / food supply - livestock and fish primary equivalent. FAO. 2015. Available at: http://faostat3.fao.org/. Accessed 3-11-2015
- 22.Wille K, Kiebooms JL, Claessens M, Rappé K, Vanden Bussche J, Noppe H, Van Praet N, De Wulf E, Van Caeter P, Janssen C, De Brabander H, Vanhaecke L. Development of analytical strategies using U-HPLC-MS/MS and LC-ToF-MS for the quantification of micropollutants in marine organisms. Anal Bioanal Chem. 2011;400(5):1459–72.Google Scholar
- 26.Galvao P, Henkelmann B, Longo R, Lailson-Brito J, Torres JPM, Schramm K-W, Malm O. Distinct bioaccumulation profile of pesticides and dioxin-like compounds by mollusk bivalves reared in polluted and unpolluted tropical bays: Consumption risk and seasonal effect. Food Chem. 2012;134(4):2040–8.CrossRefGoogle Scholar
- 31.Rejczak T, Tuzimski T. A review of recent developments and trends in the QuEChERS sample preparation approach. Open Chem 13(1):980–1010.Google Scholar
- 32.Camel V (2001) Recent extraction techniques for solid matrices-supercritical fluid extraction, pressurized fluid extraction and microwave-assisted extraction: their potential and pitfalls. Critical review-vol 7. AnalystGoogle Scholar
- 35.Commission Decision (2002/657/EC) implementing council directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Sect. 8Google Scholar
- 36.Directive 2013/39/EU of the European parliament and of the council of 12 August 2013: amending directives 2000/60/EC and 2008/105/EC as regards priority substances in the field of water policyGoogle Scholar
- 37.European Commission Directorate General Health and Consumer Protection. Guidance document on pesticides residue analytical methods. In: Directorate General Health and Consumer Protection, 2010. p. 27; SANCO/825/00 rev 8.1Google Scholar
- 41.Mouneyrac C, Linot S, Amiard JC, Amiard-Triquet C, Métais I, Durou C, Minier C, Pellerin J. Biological indices, energy reserves, steroid hormones, and sexual maturity in the infaunal bivalve Scrobicularia plana from three sites differing by their level of contamination. Gen Comp Endocr. 2008;157(2):133–41.CrossRefGoogle Scholar
- 42.Anastassiades M, Lehotay SJ, Tajnbaher D, Schenck FJ. 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. 2003;86(2):412–31.Google Scholar
- 43.AOAC International. Pesticide residues in foods by acetonitrile extraction and partitioning with magnesium sulfate. Official methods of analysis 200701. 2007;AOAC 2007.01,:9Google Scholar
- 46.Hammer Ø, Harper D, Ryan P. PAST–Paleontological statistics software package for education and data analysis, version. 1.73. Paleontologia Electronica. 2001;4:1–9.Google Scholar
- 47.Muzyka A, Bazna A, Semilekto Y, Yemelyanov A (2012) Prism 6 for Windowns. 6.01 edition. GraphPad Software, Inc.: San Diego, CAGoogle Scholar
- 48.Mendes de Oliveira MCR (2012) [Moluscos bivalves em Portugal: composição química e metais contaminantes.] Universidade Nova de Lisboa: LisboaGoogle Scholar
- 51.Payá P, Anastassiades M, Mack D, Sigalova I, Tasdelen B, Oliva J, Barba A. Analysis of pesticide residues using the Quick Easy Cheap Effective Rugged and Safe (QuEChERS) pesticide multiresidue method in combination with gas and liquid chromatography and tandem mass spectrometric detection. Anal Bioanal Chem. 2007;389(6):1697–714.CrossRefGoogle Scholar
- 52.Čajka T, Maštovská K, Lehotay SJ, Hajšlová J. Use of automated direct sample introduction with analyte protectants in the GC-MS analysis of pesticide residues. J Sep Sci. 2005;28(9/10):1048–60.Google Scholar
- 59.Damásio J, Navarro-Ortega A, Tauler R, Lacorte S, Barceló D, Soares A, López M, Riva M, Barata C. Identifying major pesticides affecting bivalve species exposed to agricultural pollution using multi-biomarker and multivariate methods. Ecotoxicology. 2010;19(6):1084–94.Google Scholar
- 64.Machado L, Bebianno M, Boski T, Moura D. Trace metals on the Algarve coast. 2: Bioaccumulation in mussels Mytilus galloprovincialis (Lamarck, 1819). Bol Inst Esp Oceanogr. 1999;15(1):465–71.Google Scholar