Skip to main content
Log in

Development of multi-residue extraction procedures using QuEChERS and liquid chromatography tandem mass spectrometry for the determination of different types of organic pollutants in mussel

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


This study aimed to develop multi-residue methods for the extraction of organic pollutants in mussels (Mytilus galloprovincialis), including 11 pharmaceuticals, 5 pesticides, 5 perfluoroalkyl substances (PFASs) and 2 illicit drugs. The combination of 4 different QuEChERS methods and 12 clean-ups (a total of 44 combinations) was tested. QuEChERS included acidified (AQ), non-acidified (SQ) and their miniaturized versions. The clean-ups included 6 different conventional dispersive solid phase extraction (dSPE) plus 2 enhanced matrix removal (EMR-Lipid) and 4 SPE procedures (including sorbents focused on phospholipid removal and polymer-based). After sample analysis via HPLC-MS/MS, the three methods that provided the best results were validated in terms of linearity, accuracy, precision, sensitivity and matrix effect. The methods selected were the combination of (i) SQ and EMR-Lipid, (ii) AQ and Z-sep+ bulk-based dSPE and (iii) AQ and graphitized carbon black (GCB)-based dSPE. Recoveries at two concentration levels (50 and 500 ng/g) ranged 54–124%, 59–124% and 60–127%, respectively, and limits of quantification (LOQs) were < 30 ng/g for most analytes using any of the methods. The three methods were tested in non-spiked mussel samples purchased in local markets, but organic pollutants were not detected in any sample. However, the methods probed to successfully extract a wide range of organic pollutants families in mussel samples from the market and from bioaccumulation trials.

Graphical abstract

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

All data generated or analysed during this study are included in this published article.

Code availability

Not applicable


  1. Campo J, Picó Y. Emerging contaminants. Compr Anal Chem. 2015;515–78.

  2. Campo J, Lorenzo M, Pérez F, Picó Y, Farré ML, Barceló D. Analysis of the presence of perfluoroalkyl substances in water, sediment and biota of the Jucar River (E Spain). Sources, partitioning and relationships with water physical characteristics. Environ Res. 2016;147:503–12.

    Article  CAS  Google Scholar 

  3. Ccanccapa A, Masiá A, Navarro-Ortega A, Picó Y, Barceló D. Pesticides in the Ebro River basin: occurrence and risk assessment. Environ Pollut. 2016;211:414–24.

    Article  CAS  Google Scholar 

  4. Sadutto D, Picó Y. Sample preparation to determine pharmaceutical and personal care products in an all-water matrix: solid phase extraction. Molecules (Basel, Switzerland). 2020;25(21).

  5. Guo X, Feng C. Biological toxicity response of Asian clam (Corbicula fluminea) to pollutants in surface water and sediment. Sci Total Environ. 2018;631-632:56–70.

    Article  CAS  Google Scholar 

  6. Álvarez-Ruiz R, Picó Y. Analysis of emerging and related pollutants in aquatic biota. Trends Environ Anal Chem. 2020;25:e00082.

    Article  Google Scholar 

  7. Fu L, Lu X, Tan J, Wang L, Chen J. Multiresidue determination and potential risks of emerging pesticides in aquatic products from Northeast China by LC–MS/MS. J Environ Sci. 2018;63:116–25.

    Article  Google Scholar 

  8. Pico Y, Belenguer V, Corcellas C, Diaz-Cruz MS, Eljarrat E, Farré M, et al. Contaminants of emerging concern in freshwater fish from four Spanish rivers. Sci Total Environ. 2019;659:1186–98.

    Article  CAS  Google Scholar 

  9. Belenguer V, Martinez-Capel F, Masiá A, Picó Y. Patterns of presence and concentration of pesticides in fish and waters of the Júcar River (Eastern Spain). J Hazard Mater. 2014;265:271–9.

    Article  CAS  Google Scholar 

  10. Kaczyński P, Łozowicka B, Perkowski M, Szabuńko J. Multiclass pesticide residue analysis in fish muscle and liver on one-step extraction-cleanup strategy coupled with liquid chromatography tandem mass spectrometry. Ecotoxicol Environ Saf. 2017;138:179–89.

    Article  Google Scholar 

  11. Gao Y, Zhang Q, Li X, Li X, Li H. Simultaneous determination of legacy and emerging per- and polyfluoroalkyl substances in fish by QuEChERS coupled with ultrahigh performance liquid chromatography tandem mass spectrometry. Anal Methods. 2018;10(47):5715–22.

    Article  CAS  Google Scholar 

  12. López-García E, Postigo C, López de Alda M. Psychoactive substances in mussels: analysis and occurrence assessment. Mar Pollut Bull. 2019;146:985–92.

    Article  Google Scholar 

  13. Belitz H-D, Schieberle WGP. Food chemistry: Springer Science & Business Media; 2013.

  14. Seval D, Abdullah Ö, Meryem YÇ, İsmihan K, Sedat K. The fatty acid composition of cultured mussels (Mytilus galloprovincialis Lamarck 1819) in offshore longline system in the Black Sea. J Aquac Mar Biol. 2015:2(6).

  15. Donato P, Micalizzi G, Oteri M, Rigano F, Sciarrone D, Dugo P, et al. Comprehensive lipid profiling in the Mediterranean mussel (Mytilus galloprovincialis) using hyphenated and multidimensional chromatography techniques coupled to mass spectrometry detection. Anal Bioanal Chem. 2018;410(14):3297–313.

    Article  CAS  Google Scholar 

  16. Prato E, Danieli A, Maffia M, Biandolino F. Lipid and fatty acid compositions of Mytilus galloprovincialis cultured in the Mar Grande of Taranto (Southern Italy): feeding strategies and trophic relationships. Zool Stud. 2010;49(2):211–9.

    CAS  Google Scholar 

  17. Anastassiades M, Lehotay SJ, Štajnbaher 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.

    Article  CAS  Google Scholar 

  18. Jesús F, Hladki R, Gérez N, Besil N, Niell S, Fernández G, et al. Miniaturized QuEChERS based methodology for multiresidue determination of pesticides in odonate nymphs as ecosystem biomonitors. Talanta. 2018;178:410–8.

    Article  Google Scholar 

  19. Peña-Herrera JM, Montemurro N, Barceló D, Pérez S. Development and validation of an analytical method for determination of pharmaceuticals in fish muscle based on QuEChERS extraction and SWATH acquisition using LC-QTOF-MS/MS system. Talanta. 2019;199:370–9.

    Article  Google Scholar 

  20. Gadelha JR, Rocha AC, Camacho C, Eljarrat E, Peris A, Aminot Y, et al. Persistent and emerging pollutants assessment on aquaculture oysters (Crassostrea gigas) from NW Portuguese coast (Ria De Aveiro). Sci Total Environ. 2019;666:731–42.

    Article  CAS  Google Scholar 

  21. Baduel C, Mueller JF, Tsai H, Gomez Ramos MJ. Development of sample extraction and clean-up strategies for target and non-target analysis of environmental contaminants in biological matrices. J Chromatogr. 2015;1426:33–47.

    Article  CAS  Google Scholar 

  22. Huerta B, Jakimska A, Llorca M, Ruhí A, Margoutidis G, Acuña V, et al. Development of an extraction and purification method for the determination of multi-class pharmaceuticals and endocrine disruptors in freshwater invertebrates. Talanta. 2015;132:373–81.

    Article  CAS  Google Scholar 

  23. Carmona E, Andreu V, Picó Y. Multi-residue determination of 47 organic compounds in water, soil, sediment and fish—Turia River as case study. J Pharm Biomed Anal. 2017;146:117–25.

    Article  CAS  Google Scholar 

  24. Andrés-Costa MJ, Pascual-Aguilar J, Andreu V, Picó Y. Assessing drugs of abuse distribution in Turia River based on geographic information system and liquid chromatography mass spectrometry. Sci Total Environ. 2017;609:360–9.

    Article  Google Scholar 

  25. European Standards CSN EN 15662. Foods of plant origin - multimethod for the determination of pesticide residues using GC- and LC-based analysis following acetonitrile extraction/partitioning and clean-up by dispersive SPE - modular QuEChERS-method. 2018.

  26. AOAC-International. Pesticides residues in foods by acetonitrile extraction and partitioning with magnesion sulfate. 2007.

  27. Phenomenex. [cited 2020 July].

  28. Smith GA, Zaffiro AD, Zimmerman ML, Munch DJ. Method 539: Determination of hormones in drinking water by solid phase extraction (SPE) and liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). US EPA. 2010.

  29. Guidance document on analytical quality control and method validation procedures for pesticide residues and analysis in food and feed. SANTE/12682/2019. European Commission. 2019.

  30. Sadutto D, Álvarez-Ruiz R, Picó Y. Systematic assessment of extraction of pharmaceuticals and personal care products in water and sediment followed by liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem. 2020;412(1):113–27.

    Article  CAS  Google Scholar 

  31. Núñez M, Borrull F, Fontanals N, Pocurull E. Determination of pharmaceuticals in bivalves using QuEChERS extraction and liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem. 2015;407(13):3841–9.

    Article  Google Scholar 

  32. METLIN. [cited 2020 August].

  33. Sznajder-Katarzyńska K, Surma M, Wiczkowski W, Cieślik E. The perfluoroalkyl substance (PFAS) contamination level in milk and milk products in Poland. Int Dairy J. 2019;96:73–84.

    Article  Google Scholar 

  34. Genualdi S, Jeong N, de Jager L, Begley T. Investigation into perfluoroalkyl substances (PFASs) in a cranberry bog: method development and sampling results. Food . Contam Part A. 2017;34(12):2181–9.

    Article  CAS  Google Scholar 

  35. Tian F, Qiao C, Luo J, Guo L, Pang T, Pang R, et al. Development of a fast multi-residue method for the determination of succinate dehydrogenase inhibitor fungicides in cereals, vegetables and fruits by modified QuEChERS and UHPLC-MS/MS. J Chromatogr B. 2020;1152:122261.

    Article  CAS  Google Scholar 

  36. Sapozhnikova Y, Lehotay SJ. Multi-class, multi-residue analysis of pesticides, polychlorinated biphenyls, polycyclic aromatic hydrocarbons, polybrominated diphenyl ethers and novel flame retardants in fish using fast, low-pressure gas chromatography–tandem mass spectrometry. Anal Chim Acta. 2013;758:80–92.

    Article  CAS  Google Scholar 

  37. Hussain S, Schönbichler SA, Güzel Y, Sonderegger H, Abel G, Rainer M, et al. Solid-phase extraction of galloyl- and caffeoylquinic acids from natural sources (Galphimia glauca and Arnicae flos) using pure zirconium silicate and bismuth citrate powders as sorbents inside micro spin columns. J Pharm Biomed Anal. 2013;84:148–58.

    Article  CAS  Google Scholar 

  38. Agilent. QuEChERS Enhanced Matrix Removal-Lipid [cited 2020 November].

  39. Munaretto JS, May MM, Saibt N, Zanella R. Liquid chromatography with high resolution mass spectrometry for identification of organic contaminants in fish fillet: screening and quantification assessment using two scan modes for data acquisition. J Chromatogr. 2016;1456:205–16.

    Article  CAS  Google Scholar 

  40. Chiesa LM, Nobile M, Malandra R, Pessina D, Panseri S, Labella GF, et al. Food safety traits of mussels and clams: distribution of PCBs, PBDEs, OCPs, PAHs and PFASs in sample from different areas using HRMS-Orbitrap® and modified QuEChERS extraction followed by GC-MS/MS. Food Addit Contam Part A. 2018;35(5):959–71.

    Article  CAS  Google Scholar 

  41. Berlioz-Barbier A, Buleté A, Faburé J, Garric J, Cren-Olivé C, Vulliet E. Multi-residue analysis of emerging pollutants in benthic invertebrates by modified micro-quick-easy-cheap-efficient-rugged-safe extraction and nanoliquid chromatography–nanospray–tandem mass spectrometry analysis. J Chromatogr. 2014;1367:16–32.

    Article  CAS  Google Scholar 

  42. Han L, Matarrita J, Sapozhnikova Y, Lehotay SJ. Evaluation of a recent product to remove lipids and other matrix co-extractives in the analysis of pesticide residues and environmental contaminants in foods. J Chromatogr. 2016;1449:17–29.

    Article  CAS  Google Scholar 

  43. Zhang H, Wang J, Li L, Wang Y. Determination of 103 pesticides and their main metabolites in animal origin food by QuEChERS and liquid chromatography–tandem mass spectrometry. Food Anal Methods. 2017;10(6):1826–43.

    Article  Google Scholar 

Download references


We thank the mass spectrometry section of the Central Services of Support to the Experimental Research (SCSIE) of the Universitat de València for providing us access to QTRAP 6500 (Applied Biosciences), and to Dr. Sales Galletero for her help. We also thank Phenomenex® and especially Noemi Fillol for providing the SPE cartridges employed for the extraction procedures.


The research that led to these results received funding from the Spanish Ministry of Science, Innovation and Universities and the ERDF (European Regional Development Fund) through the project WETANDPAC (RTI2018-097158-B-C31) and from the Generalitat Valenciana through the project ANTROPOCEN@ (PROMETEO/2018/155). R. Álvarez-Ruiz acknowledges the Spanish Ministry of Science, Innovation and Universities and the ERDF for his FPI grant BES-2016-078612.

Author information

Authors and Affiliations



Conceptualization: Rodrigo Álvarez-Ruiz, Yolanda Picó; methodology: Rodrigo Álvarez-Ruiz, Daniele Sadutto; formal analysis and investigation: Rodrigo Álvarez-Ruiz, Daniele Sadutto; writing—original draft preparation: Rodrigo Álvarez-Ruiz, Julián Campo; writing—review and editing: Yolanda Picó, Julián Campo, Rodrigo Álvarez-Ruiz, Daniele Sadutto; funding acquisition: Yolanda Picó; resources: Yolanda Picó; supervision: Julián Campo.

Corresponding author

Correspondence to Rodrigo Álvarez-Ruiz.

Ethics declarations

Ethics approval

Not applicable

Consent to participate

Not applicable

Consent for publication

Not applicable

Conflict of interest

The authors declare no competing interests.

Source of biological material

Local supermarkets from the city of Valencia, Spain.

Additional information

Publisher’s note

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

Supplementary information


(DOCX 522 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Álvarez-Ruiz, R., Picó, Y., Sadutto, D. et al. Development of multi-residue extraction procedures using QuEChERS and liquid chromatography tandem mass spectrometry for the determination of different types of organic pollutants in mussel. Anal Bioanal Chem 413, 4063–4076 (2021).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: