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Centrifugeless dispersive liquid-liquid microextraction based on salting-out phenomenon as an efficient method for determination of phenolic compounds in environmental samples


A new centrifugeless dispersive liquid-liquid microextraction (DLLME) method was applied for the convenient extraction of some phenolic compounds from environmental samples. After dispersing the extracting solvent into the sample solution (10.0 mL), the mixture was passed through a small column filled with 5 g sodium chloride. As a result, phase separation was achieved via the salting-out phenomenon, and the extracting solvent was suspended on top of the sample solution. Using a low-toxic and solidifiable extracting solvent (1-dodecanol), after immersing the column into an ice bath, the extracting solvent was solidified, collected easily, and injected into an HPLC-UV instrument. The overall extraction time was 7 min, consumption of the extracting solvent was efficiently reduced to 50 μL, and the centrifugation step was simply eliminated, which made the automation of the procedure easier than the normal DLLME technique. A series of parameters influencing the extraction were investigated systematically. The optimal experimental conditions were found to be 50 μL of 1-dodecanol as the extracting solvent, a flow rate of 2.0 mL min−1, and a pH value of 4.0 for the sample solution. Under these conditions, the method provided a good linearity in the range of 0.5–800 ng mL−1, low limits of detection (0.1–0.3 ng mL−1), good extraction repeatabilities (RSDs below 9.1%, n = 5), and enrichment factors of 100–160.

Schematic diagram of the centrifugeless dispersive liquid-liquid microextraction

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  1. 1.

    Junk GA, Richard JJ. Organics in water: solid phase extraction on a small scale. Anal Chem. 1988;60(5):451–4.

    CAS  Article  Google Scholar 

  2. 2.

    Karlberg B, Thelander S. Extraction based on the flow-injection principle. Anal Chim Acta. 1978;98(1):1–7.

    CAS  Article  Google Scholar 

  3. 3.

    Ndiomu DP, Simpson CF. Some applications of supercritical fluid extraction. Anal Chim Acta. 1988;213:237–43.

    CAS  Article  Google Scholar 

  4. 4.

    Ganzler K, Salgó A, Valkó K. Microwave extraction. J Chromatogr A. 1986;371:299–306.

    CAS  Article  Google Scholar 

  5. 5.

    Risticevic S, Niri VH, Vuckovic D, Pawliszyn J. Recent developments in solid-phase microextraction. Anal Bioanal Chem. 2009;393(3):781–95.

    CAS  Article  Google Scholar 

  6. 6.

    He Y, Lee HK. Liquid-phase microextraction in a single drop of organic solvent by using a conventional microsyringe. Anal Chem. 1997;69(22):4634–40.

    CAS  Article  Google Scholar 

  7. 7.

    Jeannot MA, Cantwell FF. Solvent microextraction into a single drop. Anal Chem. 1996;68(13):2236–40.

    CAS  Article  Google Scholar 

  8. 8.

    Nerín C, Salafranca J, Aznar M, Batlle R. Critical review on recent developments in solventless techniques for extraction of analytes. Anal Bioanal Chem. 2008;393(3):809–33.

    Article  Google Scholar 

  9. 9.

    Rasmussen KE, Pedersen-Bjergaard S. Developments in hollow fibre-based, liquid-phase microextraction. Trends Anal Chem. 2004;23(1):1–10.

    CAS  Article  Google Scholar 

  10. 10.

    Pedersen-Bjergaard S, Rasmussen KE. Liquid–liquid–liquid microextraction for sample preparation of biological fluids prior to capillary electrophoresis. Anal Chem. 1999;71(14):2650–6.

    CAS  Article  Google Scholar 

  11. 11.

    Pedersen-Bjergaard S, Rasmussen KE. Electrokinetic migration across artificial liquid membranes: new concept for rapid sample preparation of biological fluids. J Chromatogr A. 2006;1109(2):183–90.

    CAS  Article  Google Scholar 

  12. 12.

    Bazregar M, Rajabi M, Yamini Y, Asghari A, Abdossalami asl Y. In-tube electro-membrane extraction with a sub-microliter organic solvent consumption as an efficient technique for synthetic food dyes determination in foodstuff samples. J Chromatogr A. 2015;1410:35–43.

    CAS  Article  Google Scholar 

  13. 13.

    Ramos-Payán M, Villar-Navarro M, Fernández-Torres R, Callejón-Mochón M, Bello-López MÁ. Electromembrane extraction (EME)—an easy, novel and rapid extraction procedure for the HPLC determination of fluoroquinolones in wastewater samples. Anal Bioanal Chem. 2013;405(8):2575–84.

    Article  Google Scholar 

  14. 14.

    Berijani S, Assadi Y, Anbia M, Milani Hosseini M-R, Aghaee E. Dispersive liquid–liquid microextraction combined with gas chromatography-flame photometric detection: very simple, rapid and sensitive method for the determination of organophosphorus pesticides in water. J Chromatogr A. 2006;1123(1):1–9.

    CAS  Article  Google Scholar 

  15. 15.

    Viñas P, Campillo N, López-García I, Hernández-Córdoba M. Dispersive liquid–liquid microextraction in food analysis. A critical review. Anal Bioanal Chem. 2014;406(8):2067–99.

    Article  Google Scholar 

  16. 16.

    Ghambarian M, Yamini Y, Esrafili A, Yazdanfar N, Moradi M. A new concept of hollow fiber liquid–liquid–liquid microextraction compatible with gas chromatography based on two immiscible organic solvents. J Chromatogr A. 2010;1217(36):5652–8.

    CAS  Article  Google Scholar 

  17. 17.

    Hemmati M, Asghari A, Bazregar M, Rajabi M. Rapid determination of some beta-blockers in complicated matrices by tandem dispersive liquid-liquid microextraction followed by high performance liquid chromatography. Anal Bioanal Chem. 2016;408(28):8163–76.

    CAS  Article  Google Scholar 

  18. 18.

    Kocúrová L, Balogh IS, Šandrejová J, Andruch V. Recent advances in dispersive liquid–liquid microextraction using organic solvents lighter than water. A review. Microchem J. 2012;102:11–7.

    Article  Google Scholar 

  19. 19.

    Bazregar M, Rajabi M, Yamini Y, Asghari A, Hemmati M. Tandem air-agitated liquid–liquid microextraction as an efficient method for determination of acidic drugs in complicated matrices. Anal Chim Acta. 2016;917:44–52.

    CAS  Article  Google Scholar 

  20. 20.

    Bazregar M, Rajabi M, Yamini Y, Saffarzadeh Z, Asghari A. Tandem dispersive liquid–liquid microextraction as an efficient method for determination of basic drugs in complicated matrices. J Chromatogr A. 2016;1429:13–21.

    CAS  Article  Google Scholar 

  21. 21.

    Farajzadeh MA, Mogaddam MRA. Air-assisted liquid–liquid microextraction method as a novel microextraction technique; application in extraction and preconcentration of phthalate esters in aqueous sample followed by gas chromatography–flame ionization detection. Anal Chim Acta. 2012;728:31–8.

    CAS  Article  Google Scholar 

  22. 22.

    Maya F, Estela JM, Cerdà V. Completely automated in-syringe dispersive liquid–liquid microextraction using solvents lighter than water. Anal Bioanal Chem. 2012;402(3):1383–8.

    CAS  Article  Google Scholar 

  23. 23.

    Wu Q, Zhou X, Li Y, Zang X, Wang C, Wang Z. Application of dispersive liquid–liquid microextraction combined with high-performance liquid chromatography to the determination of carbamate pesticides in water samples. Anal Bioanal Chem. 2009;393(6):1755–61.

    CAS  Article  Google Scholar 

  24. 24.

    Ebrahimpour B, Yamini Y, Esrafili A. Emulsification liquid phase microextraction followed by on-line phase separation coupled to high performance liquid chromatography. Anal Chim Acta. 2012;751:79–85.

    CAS  Article  Google Scholar 

  25. 25.

    Sarafraz-Yazdi A, Amiri A. Liquid-phase microextraction. Trends Anal Chem. 2010;29(1):1–14.

    CAS  Article  Google Scholar 

  26. 26.

    Khalili Zanjani MR, Yamini Y, Shariati S, Jönsson JÅ. A new liquid-phase microextraction method based on solidification of floating organic drop. Anal Chim Acta. 2007;585(2):286–93.

    CAS  Article  Google Scholar 

  27. 27.

    Rodríguez I, Llompart MP, Cela R. Solid-phase extraction of phenols. J Chromatogr A. 2000;885(1–2):291–304.

    Article  Google Scholar 

  28. 28.

    Rodríguez I, Turnes MI, Mejuto MC, Cela R. Determination of chlorophenols at the sub-ppb level in tap water using derivatization, solid-phase extraction and gas chromatography with plasma atomic emission detection. J Chromatogr A. 1996;721(2):297–304.

    Article  Google Scholar 

  29. 29.

    Lin C-Y, Huang S-D. Application of liquid–liquid–liquid microextraction and ion-pair liquid chromatography coupled with photodiode array detection for the determination of chlorophenols in water. J Chromatogr A. 2008;1193(1–2):79–84.

    CAS  Article  Google Scholar 

  30. 30.

    Sulej-Suchomska AM, Polkowska Z, Chmiel T, Dymerski TM, Kokot ZJ, Namiesnik J. Solid phase microextraction-comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry: a new tool for determining PAHs in airport runoff water samples. Anal Methods. 2016;8(22):4509–20.

    CAS  Article  Google Scholar 

  31. 31.

    Liu Q, Shi J, Zeng L, Wang T, Cai Y, Jiang G. Evaluation of graphene as an advantageous adsorbent for solid-phase extraction with chlorophenols as model analytes. J Chromatogr A. 2011;1218(2):197–204.

    CAS  Article  Google Scholar 

  32. 32.

    Santos FJ, Jáuregui O, Pinto FJ, Galceran MT. Experimental design approach for the optimization of supercritical fluid extraction of chlorophenols from polluted soils. J Chromatogr A. 1998;823(1–2):249–58.

    CAS  Article  Google Scholar 

  33. 33.

    Moradi M, Yamini Y, Esrafili A, Seidi S. Application of surfactant assisted dispersive liquid–liquid microextraction for sample preparation of chlorophenols in water samples. Talanta. 2010;82(5):1864–9.

    CAS  Article  Google Scholar 

  34. 34.

    Calvo Seronero L, Fernández Laespada ME, Luis Pérez Pavón J, Moreno Cordero B. Cloud point preconcentration of rather polar compounds: application to the high-performance liquid chromatographic determination of priority pollutant chlorophenols. J Chromatogr A. 2000;897(1–2):171–6.

    Article  Google Scholar 

  35. 35.

    Fattahi N, Assadi Y, Hosseini MRM, Jahromi EZ. Determination of chlorophenols in water samples using simultaneous dispersive liquid–liquid microextraction and derivatization followed by gas chromatography-electron-capture detection. J Chromatogr A. 2007;1157(1–2):23–9.

    CAS  Article  Google Scholar 

  36. 36.

    Santana CM, Padrón MET, Ferrera ZS, Rodríguez JJS. Development of a solid-phase microextraction method with micellar desorption for the determination of chlorophenols in water samples: comparison with conventional solid-phase microextraction method. J Chromatogr A. 2007;1140(1–2):13–20.

    Article  Google Scholar 

  37. 37.

    López-Jiménez FJ, Rubio S, Pérez-Bendito D. Single-drop coacervative microextraction of organic compounds prior to liquid chromatography: theoretical and practical considerations. J Chromatogr A. 2008;1195(1–2):25–33.

    Article  Google Scholar 

  38. 38.

    Fan C, Li N, Cao X. Determination of chlorophenols in honey samples using in-situ ionic liquid-dispersive liquid–liquid microextraction as a pretreatment method followed by high-performance liquid chromatography. Food Chem. 2015;174:446–51.

    CAS  Article  Google Scholar 

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The authors would like to thank the Semnan University Research Council for the financial support of this work.

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Correspondence to Maryam Rajabi.

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Mirparizi, E., Rajabi, M., Bazregar, M. et al. Centrifugeless dispersive liquid-liquid microextraction based on salting-out phenomenon as an efficient method for determination of phenolic compounds in environmental samples. Anal Bioanal Chem 409, 3007–3016 (2017).

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  • Centrifugeless dispersive liquid-liquid microextraction
  • Solidification of floating organic droplets
  • Phenolic compounds
  • Salting-out phenomenon