, Volume 82, Issue 1, pp 235–250 | Cite as

Possibilities and Limitations of Isocratic Fast Liquid Chromatography-Tandem Mass Spectrometry Analysis of Pesticide Residues in Fruits and Vegetables

  • Steven J. LehotayEmail author
Part of the following topical collections:
  1. 50th Anniversary Commemorative Issue


Currently, state-of-the-art analytical methods for multiclass, multiresidue monitoring of pesticides in foods use ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for analysis of LC-amenable analytes. UHPLC-MS/MS for > 100 pesticides typically takes 10 min per injection using gradient elution in the reversed-phase mode, plus typically 3–5 min for re-equilibration between injections. Isocratic mobile-phase conditions eliminate need for the re-equilibration, and can greatly speed analysis time. In this study, fast isocratic LC-MS/MS was evaluated using a C18 analytical column of 3 cm × 4.6 mm i.d. with 3 µm particles and a mobile phase consisting of 10 mM ammonium formate at pH 3 in 47.5/47.5/5 (v/v/v) acetonitrile/methanol/water. Flow rate was 0.4 mL/min and injection volume was 50 µL. Sample preparation entailed a formate-buffered QuEChERS method for fruit and vegetable samples, typically yielding extract of 1.27 g/mL sample equivalent in 94/6 acetonitrile/water without additional cleanup. The analysis time was 2.6 min covering 88 diverse pesticide analytes each with three ion transitions (dwell times of 5 ms and 5 ms interscan delays). Validation experiments involving fortification of water, pear, tomato, cucumber, eggplant, and cilantro at 10 and 100 ng/g (n = 10 for each matrix and level) showed that the method achieved acceptable quantification with 70–120% recoveries and ≤ 25% RSD for 32–62 (36–70%) of the analytes depending on the matrix. Using regulatory identification criteria, only 6 false positives occurred above 10 ng/g among 4400 analyte/matrix/sample combinations, but false negatives varied depending on the pesticide/matrix pair, with results improving significantly for analytes with retention times > 1.3 min. This study demonstrated the feasibility and limits of isocratic LC-MS/MS for rapid screening of common commodities monitored for pesticide residues.

Graphical Abstract


Isocratic liquid chromatography-tandem mass spectrometry (LC-MS/MS) High-throughput analysis QuEChERS sample preparation Qualitative identification Pesticide residues Fruits and vegetables 



The author thanks Robyn Moten for technical assistance in the laboratory and Gary Strahan for NMR analysis of the extracts to determine water content.


Mention of brand or firm names does not constitute an endorsement by the U.S. Department of Agriculture above others of a similar nature not mentioned.

Compliance with Ethical Standards

Conflict of interest

The author declares no conflict of interest.


  1. 1.
    US Government Accountability Office (GAO). Food Safety FDA and USDA should strengthen pesticide residue monitoring programs and further disclose monitoring limitations. GAO-15-38. 2014, pp 105Google Scholar
  2. 2.
    Lehotay SJ, Chen Y (2018) Hits and misses in research trends to monitor contaminants in foods. Anal Bioanal Chem 410:5331–5351CrossRefGoogle Scholar
  3. 3.
    Stachniuk A, Fornal E (2016) Liquid chromatography-mass spectrometry in the analysis of pesticide residues in food. Food Anal Methods 9:1654–1665CrossRefGoogle Scholar
  4. 4.
    González-Curbelo M, Socas-Rodríguez B, Herrera-Herrera AV, González-Sálamo J, Hernández-Borges J, Rodríguez-Delgado M (2015) Evolution and applications of the QuEChERS method. Trends Anal Chem 71:169–185CrossRefGoogle Scholar
  5. 5.
    World Trade Organization (2018), International Trade Statistics 2015. Accessed July 2018
  6. 6.
    Strategic Consulting (2014) Food Contract Lab Report. Accessed July 2018
  7. 7.
    Ferrer C, Lozano A, Uclés S, Valverde A, Fernández-Alba AR (2017) European Union proficiency tests for pesticide residues in fruit and vegetables from 2009 to 2016: overview of the results and main achievements. Food Control 82:101–113CrossRefGoogle Scholar
  8. 8.
    Villaverde JJ, Sevilla-Morán B, López-Goti C, Alonso-Prados JL, Sandín-España P (2016) Trends in the analysis of pesticide residues to fulfil the European regulation (EC) No. 1107/2009. Trends Anal Chem 80:568–580CrossRefGoogle Scholar
  9. 9.
    Raina-Fulton R (2015) New trends in pesticide residue analysis in cereals, nutraceuticals, baby foods, and related processed consumer products. J AOAC Int 98:1163–1170CrossRefGoogle Scholar
  10. 10.
    Han L, Sapozhnikova Y, Lehotay SJ (2016) Method validation for 243 pesticides and environmental contaminants in meats and poultry by tandem mass spectrometry coupled to low-pressure gas chromatography and ultrahigh-performance liquid chromatography. Food Control 66:270–282CrossRefGoogle Scholar
  11. 11.
    Senyuva HZ, Gokmen V, Sarikaya EA (2015) Future perspectives in Orbitrap™-high-resolution mass spectrometry in food analysis: a review. Food Addit Contam A 32:1568–1606CrossRefGoogle Scholar
  12. 12.
    Wang J, Chow W, Chang J, Wong JW (2017) Development and validation of a qualitative method for target screening of 448 pesticide residues in fruits and vegetables using UHPLC/ESI Q-Orbitrap based on data-independent acquisition and compound database. J Agric Food Chem 65:473–493CrossRefGoogle Scholar
  13. 13.
    Uclés S, Uclés A, Lozano A, Martínez-Bueno MJ, Fernández-Alba AR (2017) Shifting the paradigm in gas chromatography mass spectrometry pesticide analysis using high resolution accurate mass spectrometry. J Chromatogr A 1501:107–116CrossRefGoogle Scholar
  14. 14.
    Mol HGJ, Tienstra M, Zomer P (2016) Evaluation of gas chromatography - electron ionization - full scan high resolution Orbitrap mass spectrometry for pesticide residue analysis. Anal Chim Acta 935:161–172CrossRefGoogle Scholar
  15. 15.
    Sapozhnikova Y, Lehotay SJ (2015) Review of recent developments and applications in low-pressure (vacuum outlet) gas chromatography. Anal Chim Acta 899:13–22CrossRefGoogle Scholar
  16. 16.
    Lee J, Kim L, Shin Y, Lee J, Lee J, Kim E, Moon JK, Kim JH (2017) Rapid and simultaneous analysis of 360 pesticides in brown rice, spinach, orange, and potato using microbore GC-MS/MS. J Agric Food Chem 65:3387–3395CrossRefGoogle Scholar
  17. 17.
    Nanita SC, Kaldon LG (2016) Emerging flow injection mass spectrometry methods for high-throughput quantitative analysis. Anal Bioanal Chem 408:23–33CrossRefGoogle Scholar
  18. 18.
    Nanita SC (2011) High-throughput chemical residue analysis by fast extraction and dilution flow injection mass spectrometry. Analyst 136:285–287CrossRefGoogle Scholar
  19. 19.
    Nanita SC, Kaldon LG, Bailey DL (2015) Ammonium salting out extraction with analyte preconcentration for sub-part per billion quantitative analysis in surface, ground and drinking water by flow injection tandem mass spectrometry. Anal Methods 7:2300–2312CrossRefGoogle Scholar
  20. 20.
    Mol HGJ, van Dam RCJ (2014) Rapid detection of pesticides not amenable to multi-residue methods by flow injection-tandem mass spectrometry. Anal Bioanal Chem 406:6817–6825CrossRefGoogle Scholar
  21. 21.
    Anastassiades M, Lehotay SJ, Štajnbaher 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 86:412–431Google Scholar
  22. 22.
    González-Curbelo M, Lehotay SJ, Hernández-Borges J, Rodríguez-Delgado M (2014) Use of ammonium formate in QuEChERS for high-throughput analysis of pesticides in food by fast, low-pressure gas chromatography and liquid chromatography tandem mass spectrometry. J Chromatogr A 1358:75–84CrossRefGoogle Scholar
  23. 23.
    Lehotay SJ, Sapozhnikova Y, Mol HGJ (2015) Current issues involving screening and identification of chemical contaminants in foods by mass spectrometry. Trends Anal Chem 69:62–75CrossRefGoogle Scholar
  24. 24.
    Lehotay SJ, Mastovska K, Lightfield AR, Nuñez A, Dutko T, Ng C, Bluhm L (2013) Rapid analysis of aminoglycoside antibiotics in bovine tissues using disposable pipette extraction and ultrahigh performance liquid chromatography - tandem mass spectrometry. J Chromatogr A 1313:103–112CrossRefGoogle Scholar
  25. 25.
    Dallüge J, Vreuls RJJ, van Iperen DJ, van Rijn M, Brinkman UATh (2002) Resistively heated gas chromatography coupled to quadrupole mass spectrometry. J Sep Sci 25:608:614Google Scholar
  26. 26.
    Sapozhnikova Y (2018) Development and validation of a semi-automated high-throughput analytical method for 265 pesticides and environmental contaminants in meats and poultry. J Chromatogr A. Google Scholar
  27. 27.
    Stahnke H, Kittlaus S, Kempe G, Hemmerling C, Alder L (2012) The influence of electrospray ion source design on matrix effects. J Mass Spectrom 47:875–884CrossRefGoogle Scholar
  28. 28.
    Stahnke H, Kittlaus S, Kempe G, Alder L (2012) Reduction of matrix effects in liquid chromatography-electrospray ionization-mass spectrometry by dilution of the sample extracts: how much dilution is needed? Anal Chem 84:1474–1482CrossRefGoogle Scholar
  29. 29.
    Song S, Zhu K, Han L, Sapozhnikova Y, Zhang Z, Yao W (2018) Residue analysis of 60 pesticides in red swamp crayfish using QuEChERS with high-performance liquid chromatography-tandem mass spectrometry. J Agric Food Chem 66:5031–5038CrossRefGoogle Scholar
  30. 30.
    Han L, Matarrita J, Sapozhnikova Y, Lehotay SJ (2016) 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 A 1449:17–29CrossRefGoogle Scholar
  31. 31.
    Shao G, Agar J, Giese RW (2017) Cold-induced aqueous acetonitrile phase separation: a salt-free way to begin quick, easy, cheap, effective, rugged, safe. J Chromatogr A 1506:128–133CrossRefGoogle Scholar
  32. 32.
    Lehotay SJ, Han L, Sapozhnikova Y (2016) 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 79:1113–1130CrossRefGoogle Scholar
  33. 33.
    Kwon H, Lehotay SJ, Geis-Asteggiante L (2012) Variability of matrix effects in liquid and gas chromatography—mass spectrometry analysis of pesticide residues after QuEChERS sample preparation of different food crops. J Chromatogr A 1270:235–245CrossRefGoogle Scholar

Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2018

Authors and Affiliations

  1. 1.US Department of Agriculture, Agricultural Research ServiceEastern Regional Research CenterWyndmoorUSA

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