Abstract
To prevent illegal use of clenbuterol and for quality control in the food industry, more efficient and reliable methods for clenbuterol detection are needed. In this study, clenbuterol was detected using a spectral imaging surface plasmon resonance sensor system via two inhibition methods: (1) the target site compensation method, in which anti-clenbuterol antibody was immobilized on the sensor chip as a bioprobe and (2) the solution competition method in which a clenbuterol-BSA conjugate was immobilized on the sensor chip as the bioprobe. The detectable clenbuterol concentration ranged between 6.25 and 100 μg/mL for both methods. The clenbuterol limit of detection for the target site compensation method and solution competition method are estimated to be 6.7 and 4.5 μg/mL, respectively. The proposed methods were successfully applied to the detection of clenbuterol molecules and were found to have high specificity and high-throughput and were label free and operationally convenient.
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Abbreviations
- CLEN:
-
Clenbuterol
- anti-CLEN antibody:
-
Anti-clenbuterol polyclonal antibody
- PDMS:
-
Polydimethylsiloxane
References
Roda, A., Manetta, A. C., Piazza, F., Simoni, P., & Lelli, R. (2000). A rapid and sensitive 384-microtiter wells format chemiluminescent enzyme immunoassay for clenbuterol. Talanta, 52, 311–318.
Degand, G., Bernes-Duyckaerts, A., & Maghuin-Rogister, G. (1992). Determination of clenbuterol in bovine tissues and urine by enzyme immunoassay. Journal of Agricultural and Food Chemistry, 40, 70–75.
Guo, R. X., Xu, Q., Wang, D. Y., & Hu, X. Y. (2008). Trace determination of clenbuterol with an MWCNT-Nafion nanocomposite modified electrode. Microchimica Acta, 161, 265.
Liu, L., Pan, H., Du, M., Xie, W., & Wang, J. (2010). Glassy carbon electrode modified with Nafion–Au colloids for clenbuterol electroanalysis. Electrochimica Acta, 55, 7240–7245.
Pinheiro, I., Jesuino, B., Barbosa, J., Ferreira, H., Ramos, F., Matos, J., & da Silveira, M. I. N. (2009). Clenbuterol storage stability in the bovine urine and liver samples used for European official control in the Azores Islands. Journal of Agricultural and Food Chemistry, 57, 910–914.
Pleadin, J., Vulic, A., Persi, N., Terzic, S., Andrisic, M., & Zarkovic, I. (2013). Rapid immunoassay method for the determination of clenbuterol and salbutamol in blood. Journal of Analytical Toxicology, 37, 241–245.
Ramos, F., Cristino, A., Carrola, P., Eloy, T., Silva, J. M., Castilho, M. D., & da Silveira, M. I. N. (2003). Clenbuterol food poisoning diagnosis by gas chromatography–mass spectrometric serum analysis. Analytica Chimica Acta, 483, 207.
Bomgern, A., Berggren, C., Holmberg, A., Larsson, F., Sellergren, B., & Ensing, K. (2002). Extraction of clenbuterol from calf urine using a molecularly imprinted polymer followed by quantitation by high-performance liquid chromatography with UV detection. Journal of Chromatography A, 975, 157–164.
Bardin, F., Bellemain, A., Roger, G., & Canva, M. (2009). Surface plasmon resonance spectro-imaging sensor for biomolecular surface interaction characterization. Biosensors and Bioelectronics, 24, 2100–2105.
Shankaran, D. R., Kawaguchi, T., Kim, S. J., Matsumoto, K., Toko, K., & Miura, N. (2007). Fabrication of novel molecular recognition membranes by physical adsorption and self-assembly for surface plasmon resonance detection of TNT. International Journal of Environmental Analytical Chemistry, 87, 771–781.
Gao, Y., Li, X. X., & Guo, L. H. (2012). Development of a label-free competitive ligand binding assay with human serum albumin on a molecularly engineered surface plasmon resonance sensor chip. Analytical Methods, 4, 3718.
Wood, R. (1902). On a remarkable case of uneven distribution of light in a diffraction grating spectrum. Philosophical Magazine, 4, 396–402.
Liedberg, B., Nylander, C., & Lundstrom, I. (1995). Biosensing with surface plasmon resonance—how it all started. Biosensors and Bioelectronics, 10, R1–R9.
Nylander, C., Liedberg, B., & Lind, T. (1982). Gas-detection by means of surface-plasmon resonance. Sensors and Actuators, 3, 79–88.
Homola, J., Yee, S. S., & Gauglitz, G. (1999). Surface plasmon resonance sensors: review. Sensors and Actuators B: Chemical, 54, 3–15.
Patil, S., Srinivas, S., & Jadhav, J. (2014). Evaluation of crocin and curcumin affinity on mushroom tyrosinase using surface plasmon resonance. International Journal of Biological Macromolecules, 65, 163–166.
Kwon, Y. C., Kim, M. G., Kim, E. M., Shin, Y. B., Lee, S. K., Lee, S. D., Cho, M. J., & Ro, H. S. (2011). Development of a surface plasmon resonance-based immunosensor for the rapid detection of cardiac troponin I. Biotechnology Letters, 33, 921–927.
Sakai, G., Nakata, S., Uda, T., Miura, N., & Yamazoe, N. (1999). Highly selective and sensitive SPR immunosensor for detection of methamphetamine. Electrochimica Acta, 44, 3849–3854.
Ramakrishnan, M., De Melo, F. A., Kinsey, B. M., Ladbury, J. E., Kosten, T. R., & Orson, F. M. (2012). Probing cocaine-antibody interactions in buffer and human serum. PLoS One, 7, e40518.
McNamee, S. E., Elliott, C. T., Delahaut, P., & Campbell, K. (2013). Multiplex biotoxin surface plasmon resonance method for marine biotoxins in algal and seawater samples. Environmental Science and Pollution Research, 20, 6794–6807.
Fang, X. Y., Tie, J., Xie, Y. H., Li, Q. J., Zhao, Q. C., & Fan, D. M. (2010). Detection of gastric carcinoma-associated antigen MG7-Ag in human sera using surface plasmon resonance sensor. Cancer Epidemiology, 34, 648–651.
Fang, X. Y., Liu, C. L., Cheng, X. L., Wang, Y. L., & Yang, Y. C. (2011). A spectral imaging array biosensor and its application in detection of leukemia cell. Sensors and Actuators B: Chemical, 156, 760–764.
Gambari, R., Feriotto, G., Rutigliano, C., Bianchi, N., & Mischiati, C. (2000). Biospecific interaction analysis (BIA) of low-molecular weight DNA-binding drugs. Journal of Pharmacology and Experimental Therapeutics, 294, 370–377.
Cheng, S. Y., Shi, F., Jiang, X. C., Wang, L. M., Chen, W. Q., & Zhu, C. G. (2012). Sensitive detection of small molecules by competitive immunomagnetic-proximity ligation assay. Analytical Chemistry, 84, 2129–2132.
Johansson, M. A., & Hellenas, K. E. (2004). Matrix effects in immunobiosensor determination of clenbuterol in urine and serum. Analyst, 129, 438–442.
Acknowledgments
This work was supported by NSFC (Grant No. 81371642) and the Fundamental Research Funds for the Central Universities of China.
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Wu, Y., Yao, M., Fang, X. et al. Clenbuterol Assay by Spectral Imaging Surface Plasmon Resonance Biosensor System. Appl Biochem Biotechnol 177, 1327–1337 (2015). https://doi.org/10.1007/s12010-015-1817-6
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DOI: https://doi.org/10.1007/s12010-015-1817-6