Environmental Science and Pollution Research

, Volume 22, Issue 19, pp 14882–14890 | Cite as

Simultaneous detection of imidacloprid and parathion by the dual-labeled time-resolved fluoroimmunoassay

  • Haiyan Shi
  • Enze Sheng
  • Lu Feng
  • Liangliang Zhou
  • Xiude Hua
  • Minghua Wang
Research Article


A highly sensitive direct dual-labeled time-resolved fluoroimmunoassay (TRFIA) to detect parathion and imidacloprid simultaneously in food and environmental matrices was developed. Europium (Eu3+) and samarium (Sm3+) were used as fluorescent labels by coupling separately with L1-Ab and A1P1-Ab. Under optimal assay conditions, the half-maximal inhibition concentration (IC50) and limit of detection (LOD, IC10) were 10.87 and 0.025 μg/L for parathion and 7.08 and 0.028 μg/L for imidacloprid, respectively. The cross-reactivities (CR) were negligible except for methyl-parathion (42.4 %) and imidaclothiz (103.4 %). The average recoveries of imidacloprid ranged from 78.9 to 104.2 % in water, soil, rice, tomato, and Chinese cabbage with a relative standard deviation (RSD) of 2.4 to 11.6 %, and those of parathion were from 81.5 to 110.9 % with the RSD of 3.2 to 10.5 %. The results of TRFIA for the authentic samples were validated by comparison with gas chromatography (GC) analyses, and satisfactory correlations (parathion: R 2 = 0.9918; imidacloprid: R 2 = 0.9908) were obtained. The results indicate that the dual-labeled TRFIA is convenient and reliable to detect parathion and imidacloprid simultaneously in food and environmental matrices.


Imidacloprid Parathion Europium Samarium Time-resolved fluoroimmunoassay 



This project was supported by the National Natural Science Foundation of China (31301692) and the Fundamental Research Funds for the Central Universities (KJQN201433 and KYZ201305).

Compliance with ethical standards

The manuscript has not been submitted to more than one journal for simultaneous consideration. The manuscript has not been published previously (partly or in full). A single study is not split up into several parts to increase the quantity of submissions and submitted to various journals or to one journal over time. No data have been fabricated or manipulated (including images) to support our conclusions. No data, text, or theories by others are presented as if they were the authors’ own (“plagiarism”).

All authors whose names appear on the submission have contributed sufficiently to the scientific work and therefore share collective responsibility and accountability for the results. They have reviewed this manuscript and approved to submit to your journal.

Supplementary material

11356_2015_4697_MOESM1_ESM.docx (212 kb)
Fig. S1 (DOCX 211 kb)
11356_2015_4697_MOESM2_ESM.docx (397 kb)
Fig. S2 (DOCX 397 kb)


  1. Fang S, Zhang B, Ren KW, Cao MM, Shi HY, Wang MH (2011) Development of a sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on the monoclonal antibody for the detection of the imidaclothiz residue. J Agric Food Chem 59:1594–1597CrossRefGoogle Scholar
  2. Gui WJ, Jin MJ, Sun LF, Guo YR, Zhu GN (2009) Residues determination of carbofuran in vegetables based on sensitive time-resolved fluorescence immunoassay. Food Agric Immunol 20:49–56CrossRefGoogle Scholar
  3. Hemmilä I, Dakubu S, Mukkala V, Siitari H, Lövgren T (1984) Europium as a label in time-resolved immunofluorometric assays. Anal Biochem 137:335–343CrossRefGoogle Scholar
  4. Hennion MC, Barcelo D (1998) Strengths and limitations of immunoassays for effective and efficient use for pesticide analysis in water samples: a review. Anal Chim Acta 362:3–34CrossRefGoogle Scholar
  5. Huang B, Xiao HL, Zhang J, Zhang LF, Yang HL, Zhang Y, Jin J (2009) Dual-label time-time-resolved fluoroimmunoassay for simultaneous detection of aflatoxin B1 and ochratoxin A. Arch Toxicol 83:619–624CrossRefGoogle Scholar
  6. Huang B, Zhao LL, Sun JM, Zhang J, Zhang Y, Zhang K, Jin J, Xie MH (2012) Establishment and validation of a time-resolved fluoroimmunoassay for chlorpromazine. Food Anal Methods 5:625–630CrossRefGoogle Scholar
  7. Jin RY, Guo YR, Wang CM, Wu JX, Zhu GN (2009) Development of a bispecific monoclonal antibody to pesticide carbofuran and triazophos using hybrid hybridomas. J Food Sci 74:T1–T6CrossRefGoogle Scholar
  8. Li P, Zhang Q, Zhang W (2009) Immunoassays for aflatoxins. Trends Anal Chem 28:1115–1126CrossRefGoogle Scholar
  9. Li M, Sheng EZ, Yuan YL, Liu XF, Hua XD, Wang MH (2014) Sensitive time-resolved fluoroimmunoassay for quantitative determination of clothianidin in agricultural samples. Environ Sci Pollut Res 21:5803–5809CrossRefGoogle Scholar
  10. Liu Y, Lou Y, Xu D, Qian GL, Zhang Q, Wu RR, Hu BS, Liu FQ (2009) Production and characterization of monoclonal antibody for class-specific determination of o,o-dimethyl organophosphorus pesticides and effect of heterologous coating antigens on immunoassay sensitivity. Microchemical 93:36–43CrossRefGoogle Scholar
  11. Liu ZJ, Yan X, Wang MH (2013) Development of a chemiluminescence enzyme-linked immunosorbent assay for the simultaneous detection of imidaclothiz and thiacloprid in agricultural samples. Analyst 11:3280–3286CrossRefGoogle Scholar
  12. Nuria PN, Ester GI, Ángel M, Rosa P (2007) Development of a group-specific immunoassay for sulfonamides—application to bee honey analysis. Talanta 71:923–933CrossRefGoogle Scholar
  13. Ren KW, Jin YH, Cao MM, Wang MH (2012) Detection of aryloxyphenoxypropionate herbicides by enzyme-linked immunosorbent assay. Anal Lett 45:831–840CrossRefGoogle Scholar
  14. Reverberi R, Reverberi L (2007) Factors affecting the antigen-antibody reaction. Blood Transfus 5:227–240Google Scholar
  15. Shi HY, Li HX, Hua XD, Zheng ZT, Zhu GN, Wang MH (2014) Characterization of multi-hapten antigens on antibody sensitivity and specificity for parathion. Anal Lett 47:2699–2707CrossRefGoogle Scholar
  16. Wanatabe S, Ito S, Kamata Y, Omoda N, Yamazaki T, Munakata H, Kaneko T, Yuasa Y (2001) Development of competitive enzyme-linked immunosorbent assays (ELISAs) based on monoclonal antibodies for chloronicotinoid insecticides imidacloprid and acetamiprid. Anal Chim Acta 427:211–219CrossRefGoogle Scholar
  17. Wang CM, Li XB, Liu YH, Guo YR, Xie R, Gui WJ, Zhu GN (2010) Development of a Mab-based heterologous immunoassay for the broad-selective determination of organophosphorus pesticides. J Agric Food Chem 58:5658–5663CrossRefGoogle Scholar
  18. Wu FB, Xu YY, Wang YZ, Han SQ (1999) Time-resolved fluorescence immunoassay of thyroxine in serum: immobilized antigen approach. Anal Biochem 276:171–176CrossRefGoogle Scholar
  19. Wu FB, Han SQ, Xu T, He YF (2003) Sensitive time-resolved fluoroimmunoassay for simultaneous detection of serum thyroid-stimulating hormone and total thyroxin with Eu and Sm as labels. Anal Biochem 314:87–96CrossRefGoogle Scholar
  20. Xu ZL, Dong JX, Yang JY, Wang H, Jiang YM, Lei HT, Shen YD, Sun YM (2012) Development of a sensitive time-resolved fluoroimmunoassay for organophosphorus pesticides in environmental water samples. Anal Methods 4:3484–3490CrossRefGoogle Scholar
  21. Zhang J, Guo JZ, Xiao HL (2010) Simultaneous detection of different serum pepsinogens and its primary application. World J Gastroenterol 16:3072–3077CrossRefGoogle Scholar
  22. Zhang J, Gao L, Zhou B (2011) Simultaneous detection of deoxynivalenol and zearalenone by dual-label time-resolved fluorescence immunoassay. J Sci Food Agric 91:193–197CrossRefGoogle Scholar
  23. Zhang Z, Liu JF, Feng TT, Yao Y, Gao LH, Jiang GB (2013) Time-resolved fluoroimmunoassay as an advantageous analytical method for assessing the total concentration and environmental risk of fluoroquinolones in surface waters. Environ Sci Technol 47:454–462CrossRefGoogle Scholar
  24. Zhou YL, Xia XH, Xu Y, Ke W, Yang W, Li QG (2012) Application of europium(III) chelates-bonded silica nanoparticle in time-resolved immunofluorometric detection assay for human thyroid stimulating hormone. Anal Chim Acta 722:95–99CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Haiyan Shi
    • 1
  • Enze Sheng
    • 1
  • Lu Feng
    • 1
  • Liangliang Zhou
    • 1
  • Xiude Hua
    • 1
  • Minghua Wang
    • 1
  1. 1.Department of Pesticide Science, College of Plant Protection, Jiangsu Key Laboratory of Pesticide Science, Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of EducationNanjing Agricultural UniversityNanjingPeople’s Republic of China

Personalised recommendations