Skip to main content
SpringerLink
Log in

A silica nanoparticle based 2-color immunochromatographic assay for simultaneous determination of clenbuterol and ractopamine

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

An immunochromatographic assay (ICA) is presented that can be applied to simultaneous detection of clenbuterol (CLE) and ractopamine (RAC). It is making use of two red and blue silica nanoparticles (SiNPs) that act as labels for encoding the antibodies. This design permits multiplexed analysis in a single test line and does not require an external source for photoexcitation. Anti-CLE was labeled with red SiNPs, and anti-RAC with blue SiNPs. The capture antigens CLE-BSA and RAC-BSA were placed onto the conjugate pad and the test line of the test strip, respectively. Under bare eye examination, no cross-colored lines or nonspecific bioconjugate adsorption were observed, and the visible limit of detections for CLE (red) and RAC (blue) are 3 and 2 ng‧mL−1, respectively. This design allows for multiplexed detection with reduced device dimensions and costs, and with easy integration and manufacturing. Conceivably, the method may be extended to simultaneous determination of numerous other analytes.

The principle of qualitative detection strategy of multiplex immunochromatographic assay for clenbuterol (CLE) and ractopamine (RAC) is schematically illustrated. Depending on the type and ratio of organic dyes, the color of colored silica nanoparticle can be tuned from red to purple and even to black (lower right corner).

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Scheme 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Shishani E, Chai SC, Jamokha S, Aznar G, Hoffman MK (2003) Determination of ractopamine in animal tissues by liquid chromatography-fluorescence and liquid chromatography/tandem mass spectrometry. Anal Chim Acta 483(1):137–145

    Article  CAS  Google Scholar 

  2. Strydom PE, Frylinck L, Montgomery JL, Smith MF (2009) The comparison of three β-agonists for growth performance, carcass characteristics and meat quality of feedlot cattle. Meat Sci 81(3):557–564

    Article  CAS  Google Scholar 

  3. Zhai H, Liu Z, Chen Z, Liang Z, Su Z, Wang S (2015) A sensitive electrochemical sensor with sulfonated graphene sheets/oxygen-functionalizedmulti-walled carbon nanotubes modified electrode for the detection of clenbuterol. Sensors Actuators B Chem 210:483–490

    Article  CAS  Google Scholar 

  4. Zhang LH, Wang QW, Qi Y, Li L, Wang ST, Wang XH (2019) An ultrasensitive sensor based on polyoxometalate and zirconium dioxide nanocomposites hybrids material for simultaneous detection of toxic clenbuterol and ractopamine. Sensors Actuators B-Chemical 288:347–355

    Article  CAS  Google Scholar 

  5. Fu X, Chu Y, Zhao K, Li J, Deng A (2017) Ultrasensitive detection of the beta-adrenergic agonist brombuterol by a SERS-based lateral flow immunochromatographic assay using flower-like gold-silver core-shell nanoparticles. Microchim Acta 184(6):1711–1719

    Article  CAS  Google Scholar 

  6. Wang W, Liu L, Song S, Xu L, Kuang H, Zhu J, Xu C (2017) Identification and quantification of eight Listeria monocytogene serotypes from Listeria spp. using a gold nanoparticle-based lateral flow assay. Microchim Acta 184(3):715–724

    Article  CAS  Google Scholar 

  7. Wang R, Zhang W, Wang P, Su X (2018) A paper-based competitive lateral flow immunoassay for multi beta-agonist residues by using a single monoclonal antibody labelled with red fluorescent nanoparticles. Microchim Acta 185(3)

  8. Xing C, Liu L, Song S, Feng M, Kuang H, Xu C (2015) Ultrasensitive immunochromatographic assay for the simultaneous detection of five chemicals in drinking water. Biosens Bioelectron 66:445–453

    Article  CAS  Google Scholar 

  9. Li J, Macdonald J (2016) Multiplexed lateral flow biosensors: technological advances for radically improving point-of-care diagnoses. Biosens Bioelectron 83:177–192

    Article  CAS  Google Scholar 

  10. Peng T, Wang J, Zhao S, Zeng Y, Zheng P, Liang D, Mari GM, Jiang H (2018) Highly luminescent green-emitting Au nanocluster-based multiplex lateral flow immunoassay for ultrasensitive detection of clenbuterol and ractopamine. Anal Chim Acta 1040:143–149

    Article  CAS  Google Scholar 

  11. Wang W, Su X, Ouyang H, Wang L, Fu Z (2016) A novel immunochromatographic assay based on a time-resolved chemiluminescence strategy for the multiplexed detection of ractopamine and clenbuterol. Anal Chim Acta 917:79–84

    Article  CAS  Google Scholar 

  12. Zhang M-Z, Wang M-Z, Chen Z-L, Fang J-H, Fang M-M, Liu J, Yu X-P(2009) Development of a colloidal gold-based lateral-flow immunoassay for the rapid simultaneous detection of clenbuterol and ractopamine in swine urine. Anal Bioanal Chem 395(8):2591–2599

    Article  CAS  Google Scholar 

  13. Washburn EW (1921) The dynamics of capillary flow. Phys Rev 17(3):273–283

    Article  Google Scholar 

  14. Wang C, Hou F, Ma Y (2015) Simultaneous quantitative detection of multiple tumor markers with a rapid and sensitive multicolor quantum dots based immunochromatographic test strip. Biosens Bioelectron 68:156–162

    Article  CAS  Google Scholar 

  15. Di Nardo F, Alladio E, Baggiani C, Cavalera S, Giovannoli C, Spano G, Anfossi L (2019)Colour-encoded lateral flow immunoassay for the simultaneous detection of aflatoxin B1 and type-B fumonisins in a single test line. Talanta 192:288–294

    Article  Google Scholar 

  16. Lee S, Mehta S, Erickson D (2016)Two-color lateral flow assay for multiplex detection of causative agents behind acute febrile illnesses. Anal Chem 88(17):8359–8363

    Article  CAS  Google Scholar 

  17. Di Nardo F, Baggiani C, Giovannoli C, Spano G, Anfossi L (2017) Multicolor immunochromatographic strip test based on gold nanoparticles for the determination of aflatoxin B1 and fumonisins. Microchim Acta 184(5):1295–1304

    Article  Google Scholar 

  18. Sheng W, Chang Q, Shi Y, Duan W, Zhang Y, Wang S (2018) Visual and fluorometric lateral flow immunoassay combined with a dual-functional test mode for rapid determination of tetracycline antibiotics. Microchim Acta 185(9):404

    Article  Google Scholar 

  19. Yen C-W, de Puig H, Tam JO, Gómez-Márquez J, Bosch I, Hamad-Schifferli K, Gehrke L (2015) Multicolored silver nanoparticles for multiplexed disease diagnostics: distinguishing dengue, yellow fever, and Ebola viruses. Lab Chip 15(7):1638–1641

    Article  CAS  Google Scholar 

  20. Panfilova E, Shirokov A, Khlebtsov B, Matora L, Khlebtsov N (2012) Multiplexed dot immunoassay using ag nanocubes, au/ag alloy nanoparticles, and au/ag nanocages. Nano Res 5(2):124–134

    Article  CAS  Google Scholar 

  21. Behnke T, Würth C, Hoffmann K, Hübner M, Panne U, Resch-Genger U (2011) Encapsulation of hydrophobic dyes in polystyrene micro- and nanoparticles via swelling procedures. J Fluoresc 21(3):937–944

    Article  CAS  Google Scholar 

  22. Sun Q, Zhao G, Dou W (2015) A nonenzymatic optical immunoassay strategy for detection of Salmonella infection based on blue silica nanoparticles. Anal Chim Acta 898:109–115

    Article  CAS  Google Scholar 

  23. Sun Q, Zhao G, Dou W (2015) Blue silica nanoparticle-based colorimetric immunoassay for detection of Salmonella pullorum. Anal Methods 7(20):8647–8654

    Article  CAS  Google Scholar 

  24. Yu H, Zhao G, Dou W (2015) Simultaneous detection of pathogenic Bacteria using agglutination test based on colored silica nanoparticles. Curr Pharm Biotechnol 16(8):716–723

    Article  CAS  Google Scholar 

  25. Winnik FM, Keoshkerian B, Roderick Fuller J, Hofstra PG (1990) New water-dispersible silica-based pigments: synthesis and characterization. Dyes Pigments 14(2):101–112

    Article  CAS  Google Scholar 

  26. Giesche H, Matijević E (1991)Well-defined pigments: I. monodispersed silica-acid dyes systems. Dyes Pigments 17(4):323–340

    Article  CAS  Google Scholar 

  27. Zhu C, Zhao G, Dou W (2018) A new synthesis method for bright monodispersed core-shell colored silica submicron particles. J Sol-Gel Sci Technol 85(1):76–83

    Article  CAS  Google Scholar 

  28. Zhu C, Zhao G, Dou W (2018)Core-shell red silica nanoparticles based immunochromatographic assay for detection of Escherichia coli O157:H7. Anal Chim Acta 1038:97–104

    Article  CAS  Google Scholar 

  29. Zhu C, Zhao G, Dou W (2018) Immunochromatographic assay using brightly colored silica nanoparticles as visible label for point-of-care detection of clenbuterol. Sensors Actuators B Chem 266:392–399

    Article  CAS  Google Scholar 

  30. Alhogail S, Suaifan GARY, Zourob M (2016) Rapid colorimetric sensing platform for the detection of Listeria monocytogenes foodborne pathogen. Biosens Bioelectron 86:1061–1066

    Article  CAS  Google Scholar 

  31. Shelver WL, Smith DJ (2002) Application of a monoclonal antibody-based enzyme-linked immunosorbent assay for the determination of Ractopamine in incurred samples from food animals. J Agric Food Chem 50(10):2742–2747

    Article  Google Scholar 

  32. Liu N, Su P, Gao Z, Zhu M, Yang Z, Pan X, Fang Y, Chao F (2009) Simultaneous detection for three kinds of veterinary drugs: chloramphenicol, clenbuterol and 17-beta-estradiol by high-throughput suspension array technology. Anal Chim Acta 632(1):128–134

    Article  CAS  Google Scholar 

Download references

Acknowledgments

Support by National Natural Science Foundation of Zhejiang Province (LY17C200003).

Author information

Authors and Affiliations

  1. Food Safety Key Laboratory of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018, China

    Qiongqiong Yu, Jing Liu, Guangying Zhao & Wenchao Dou

Authors
  1. Qiongqiong Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guangying Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wenchao Dou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wenchao Dou.

Ethics declarations

The author(s) declare that they have no competing interests.

Additional information

Publisher’s note

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

Electronic supplementary material

ESM 1

(DOCX 799 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yu, Q., Liu, J., Zhao, G. et al. A silica nanoparticle based 2-color immunochromatographic assay for simultaneous determination of clenbuterol and ractopamine. Microchim Acta 186, 421 (2019). https://doi.org/10.1007/s00604-019-3529-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00604-019-3529-z

Keywords

Search

Navigation