Efficient removal of ethidium bromide from aqueous solution by using DNA-loaded Fe3O4 nanoparticles

  • Zhiqiang Ge
  • Tingting Sun
  • Jinfeng Xing
  • Xuejiao Fan
Research Article


Ethidium bromide (EtBr) is widely used as DNA-staining dyes for the detection of nucleic acids in laboratories and known to be powerful mutagens and carcinogens. In the present paper, the removal of EtBr from aqueous solutions in a batch system using DNA-loaded Fe3O4 nanoparticles as a simple and efficient method was investigated. DNA was covalently loaded on the surface of Fe3O4 magnetic nanoparticles, which was confirmed by FT-IR analysis and zeta potential measurements. The morphology and crystal structure were characterized by SEM, TEM, and XRD. The influence factors on the removal efficiency such as initial EtBr concentration, contact time, adsorbent dose, pH, and temperature were also studied. The removal process of EtBr can be completed quickly within 1 min. The removal efficiency was more than 99% while the EtBr concentration was routinely used (0.5 mg L−1) in biology laboratories and the dosages of nanoparticles were 1 g L−1. For the different EtBr concentrations from 0.5 to 10 mg L−1 in aqueous solution, the goal of optimized removal was achieved by adjusting the dosage of DNA-loaded Fe3O4 nanoparticles. The optimum pH was around 7 and the operational temperature from 4 to 35 °C was appropriate. Kinetic studies confirmed that the adsorption followed second-order reaction kinetics. Thermodynamic data revealed that the process was spontaneous and exothermic. The adsorption of EtBr on DNA-loaded Fe3O4 nanoparticles fitted well with the Freundlich isotherm model. These results indicated that DNA-loaded Fe3O4 nanoparticles are a promising adsorbent for highly efficient removal of EtBr from aqueous solution in practice.


Ethidium bromide Deoxyribonucleic acid Magnetic nanoparticles Rapid removal 


Funding information

This work was supported by the National Natural Science Foundation of China (No. 31771094).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ahmadpour A, Eftekhari N, Ayati A (2014) Performance of MWCNTs and a low-cost adsorbent for chromium(VI) ion removal. J Nanostruct Chem 4:171–178CrossRefGoogle Scholar
  2. Bio-Rad Laboratories: Ethidium bromide solution, 10 mg/ml. Bio-Rad publishing Accessed 26 March 2018
  3. Carbajo J, Adan C, Rey A, Martinez-Arias A, Bahamonde A (2011) Optimization of H2O2 use during the photocatalytic degradation of ethidium bromide with TiO2 and iron-doped TiO2 catalysts. Appl Catal, B 102:85–93CrossRefGoogle Scholar
  4. Chae HS, Kim SD, Piao SH, Choi HJ (2016) Core-shell structured Fe3O4@SiO2 nanoparticles fabricated by sol-gel method and their magnetorheology. Colloid Polym Sci 294:647–655CrossRefGoogle Scholar
  5. Chen HM, Deng CH, Zhang XM (2010) Synthesis of Fe3O4@SiO2@PMMA Core-Shell-Shell magnetic microspheres for highly efficient enrichment of peptides and proteins for MALDI-ToF MS analysis. Angew Chem Int Ed 49:607–611CrossRefGoogle Scholar
  6. Chen J, Wang F, Huang K, Liu Y, Liu S (2009) Preparation of Fe3O4 nanoparticles with adjustable morphology. J Alloys Compd 475:898–902CrossRefGoogle Scholar
  7. Dai YM, Zou JQ, Liu DY, Niu LL, Zhou LL, Zhou Y, Zhang XH (2018) Preparation of Congo red functionalized Fe3O4@SiO2 nanoparticle and its application for the removal of methylene blue. Colloids Surf A Physicochem Eng Asp 550:90–98CrossRefGoogle Scholar
  8. D'Amico ML, Paiotta V, Secco F, Venturini M (2002) A kinetic study of the intercalation of ethidium bromide into poly(a)-poly(U). J Phys Chem B 106:12635–12641CrossRefGoogle Scholar
  9. Feng LY, Cao MH, Ma XY, Zhu YS, Hu CW (2012) Superparamagnetic high-surface-area Fe3O4 nanoparticles as adsorbents for arsenic removal. J Hazard Mater 217:439–446CrossRefGoogle Scholar
  10. Gupta VK, Suhas (2009) Application of low-cost adsorbents for dye removal - a review. J Environ Manag 90:2313–2342CrossRefGoogle Scholar
  11. Han S, Yu H, Yang T, Wang S, Wang X (2017) Magnetic activated-ATP@ Fe3O4 nanocomposite as an efficient Fenton-like heterogeneous catalyst for degradation of ethidium bromide. Sci Rep
  12. Hayashi M, Harada Y (2007) Direct observation of the reversible unwinding of a single DNA molecule caused by the intercalation of ethidium bromide. Nucleic Acids Res 35:e125. CrossRefGoogle Scholar
  13. Heibati B, Yetilmezsoy K, Zazouli MA, Rodriguez-Couto S, Tyagi I, Agarwal S, Gupta VK (2016) Adsorption of ethidium bromide (EtBr) from aqueous solutions by natural pumice and aluminium-coated pumice. J Mol Liq 213:41–47CrossRefGoogle Scholar
  14. Hou X, Zhao C, Tian Y, Dou S, Zhang X, Zhao J (2016) Preparation of functionalized Fe3O4@SiO2 magnetic nanoparticles for monoclonal antibody purification. Chem Res Chin Univ 32:889–894CrossRefGoogle Scholar
  15. Ivanova EP, Pham DK, Brack N, Pigram P, Nicolau DV (2004) Poly(L-lysine)-mediated immobilisation of oligonucleotides on carboxy-rich polymer surfaces. Biosens Bioelectron 19:1363–1370CrossRefGoogle Scholar
  16. Karacan P, Okay O (2013) Ethidium bromide binding to DNA cryogels. React Funct Polym 73:442–450CrossRefGoogle Scholar
  17. Liu Y (2008) New insights into pseudo-second-order kinetic equation for adsorption. Colloids Surf A Physicochem Eng Asp 320:275–278CrossRefGoogle Scholar
  18. Lunn G, Sansone EB (1987) Ethidium bromide: destruction and decontamination of solutions. Anal Biochem 162:453–458CrossRefGoogle Scholar
  19. Maurye P, Basu A, Biswas JK, Bandyopadhyay TK (2017) Electrophoresis-staining apparatus for DNA agarose gels with solution exchange and image acquisition. Instrum Sci Technol 45:49–61CrossRefGoogle Scholar
  20. Mihaiescu DE, Buteica AS, Neamtu J, Istrati D, Mindrila I (2013) Fe3O4/salicylic acid nanoparticles behavior on chick CAM vasculature. J Nanopart Res 15:1857–1867CrossRefGoogle Scholar
  21. MIT Environment, Health & Safety Office (2006): Replacing ethidium bromide in an undergraduate laboratory: SYBR safe®. MIT Pulishing education. Accessed 26 March 2018
  22. Moradi O, Fakhri A, Adami S, Adami S (2013) Isotherm, thermodynamic, kinetics, and adsorption mechanism studies of ethidium bromide by single-walled carbon nanotube and carboxylate group functionalized single-walled carbon nanotube. J Colloid Interface Sci 395:224–229CrossRefGoogle Scholar
  23. Moradi O, Norouzi M, Fakhri A, Naddafi K (2014) Interaction of removal ethidium bromide with carbon nanotube: equilibrium and isotherm studies. J Environ Health Sci Eng 12:17. CrossRefGoogle Scholar
  24. Nafisi S, Saboury AA, Keramat N, Neault J-F, Tajmir-Riahi H-A (2007) Stability and structural features of DNA intercalation with ethidium bromide, acridine orange and methylene blue. J Mol Struct 827:35–43CrossRefGoogle Scholar
  25. Phukan S, Mitra S (2012) Fluorescence behavior of ethidium bromide in homogeneous solvents and in presence of bile acid hosts. J Photochem Photobiol, A 244:9–17CrossRefGoogle Scholar
  26. Pouretedal HR, Sadegh N (2014) Effective removal of amoxicillin, cephalexin, tetracycline and penicillin G from aqueous solutions using activated carbon nanoparticles prepared from vine wood. J water. Process Eng 1:64–73Google Scholar
  27. Qiu Y, Guo H, Guo C, Zheng J, Yue T, Yuan Y (2018) One-step preparation of nano-Fe3O4 modified inactivated yeast for the adsorption of patulin. Food Control 86:310–318CrossRefGoogle Scholar
  28. Rahman MM, Lopa NS, Kim YJ, Choi D-K, Lee J-J (2016) Label-free DNA hybridization detection by poly(Thionine)-gold nanocomposite on indium tin oxide electrode. J Electrochem Soc 163:B153–B157CrossRefGoogle Scholar
  29. Ross PD, Subramanian S (1981) Thermodynamics of protein association reactions: forces contributing to stability. Biochem 20:3096–3102CrossRefGoogle Scholar
  30. Shrivas K, Ghosale A, Nirmalkar N, Srivastava A, Singh SK, Shinde SS (2017) Removal of endrin and dieldrin isomeric pesticides through stereoselective adsorption behavior on the graphene oxide-magnetic nanoparticles. Environ Sci Pollut Res 24:24980–24988CrossRefGoogle Scholar
  31. Sikora A, Bartczak D, Geissler D, Kestens V, Roebben G, Ramaye Y, Varga Z, Palmai M, Shard AG, Goenaga-Infante H, Minelli C (2015) A systematic comparison of different techniques to determine the zeta potential of silica nanoparticles in biological medium. Anal Methods 7:9835–9843CrossRefGoogle Scholar
  32. Suzuki H, Amano T, Toyooka T, Ibuki Y (2008) Preparation of DNA-adsorbed TiO2 particles with high performance for purification of chemical pollutants. Environ Sci Technol 42:8076–8082CrossRefGoogle Scholar
  33. Tao Q, Fang Y, Li T, Zhang D, Chen M, Ji S, He H, Komarneni S, Zhang H, Dong Y, Noh YD (2016) Silylation of saponite with 3-aminopropyltriethoxysilane. Appl Clay Sci 132:133–139CrossRefGoogle Scholar
  34. Vardevanyan PO, Antonyan AP, Manukyan GA, Karapetyan AT (2001) Study of ethidium bromide interaction peculiarities with DNA. Exp Mol Med 33:205–208CrossRefGoogle Scholar
  35. Vardevanyan PO, Parsadanyan MA, Antonyan AP, Hakobyan SN (2017) Thermodynamic parameters analysis of ethidium bromide and mitoxantrone binding with DNA by adsorption isotherms. Russ J Phys Chem A 91:1143–1145CrossRefGoogle Scholar
  36. Venkatesha TG, Viswanatha R, Nayaka YA, Chethana BK (2012) Kinetics and thermodynamics of reactive and vat dyes adsorption on MgO nanoparticles. Chem Eng J:198: 1–198:10CrossRefGoogle Scholar
  37. Wang J, Zheng S, Shao Y, Liu J, Xu Z, Zhu D (2010) Amino-functionalized Fe3O4@SiO2 core-shell magnetic nanomaterial as a novel adsorbent for aqueous heavy metals removal. J Colloid Interface Sci 349:293–299CrossRefGoogle Scholar
  38. Xiao L, Li J, Brougham DF, Fox EK, Feliu N, Bushmelev A, Schmidt A, Mertens N, Kiessling F, Valldor M, Fadeel B, Mathur S (2011) Water-soluble superparamagnetic magnetite nanoparticles with biocompatible coating for enhanced magnetic resonance imaging. ACS Nano 5:6315–6324CrossRefGoogle Scholar
  39. Yamada M, Kato K, Nomizu M, Sakairi N, Ohkawa K, Yamamoto H, Nishi N (2002) Preparation and characterization of DNA films induced by UV irradiation. Chem Eur J 8:1407–1412CrossRefGoogle Scholar
  40. Zare K, Gupta VK, Moradi O, Makhlou ASH, Sillanpää M, Nadagouda NM, Sadegh H, Shahryari-ghoshekandi R, Pal A, Wang ZJ, Tyagi I, Kazemi M (2015) A comparative study on the basis of adsorption capacity between CNTs and activated carbon as adsorbents for removal of noxious synthetic dyes: a review. J Nanostruct Chem 5:227–236CrossRefGoogle Scholar
  41. Zhang C, Liu L, Wang J, Rong F, Fu D (2013) Electrochemical degradation of ethidium bromide using boron-doped diamond electrode. Sep Purif Technol 107:91–101CrossRefGoogle Scholar
  42. Zhang CY, Yang LJ, Rong F, Fu DG, Gu ZZ (2012) Boron-doped diamond anodic oxidation of ethidium bromide: process optimization by response surface methodology. Electrochim Acta 64:100–109CrossRefGoogle Scholar
  43. Zhang Y, Jiao Z, Hu Y, Lv S, Fan H, Zeng Y, Hu J, Wang M (2017) Removal of tetracycline and oxytetracycline from water by magnetic Fe3O4@graphene. Environ Sci Pollut Res 24:2987–2995CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Key Laboratory of Systems Bioengineering (Ministry of Education)Tianjin UniversityTianjinPeople’s Republic of China

Personalised recommendations