Synthesis of Hapten for Indirect Competitive Immunoassay for Measuring 3,5,6-trichloro-2-pyridinol



Kim, Areumnuri;Kim, Joong-Young;Jeong, Sang-Hee;Cho, Myung-Haing;Park, Kyung-Hun;Cho, Namjun;Paik, Min Kyoung

  • 투고 : 2015.09.10
  • 심사 : 2015.10.24
  • 발행 : 2015.12.31


BACKGROUND: In this study, we have attempted to identify a urinary biomarker to assess chlorpyrifos exposure in farmers. The major metabolite and the excretion pathway of chlorpyrifos is 3,5,6-trichloro-2-pyridinol (TCP) in urine. Herein, we describe an adequate synthetic method for TCP hapten for measuring urinary TCP of farmers. METHODS AND RESULTS: First, TCP was prepared by spacer attachment through hydrolysis of thiophosphate ester from chlorpyrifos. After reaction with benzyl bromide, the TCP was transformed into 2,3,5-trichloro-6-benzyloxypyridine. Next, the chlorine in the 2 nd position of the pyridyl ring was substituted into 3-mercaptopropanoic acid spacer arm. Finally, the phenyl group attached to the 6 th position in pyridyl ring was removed for producing the targeted product, 3-(3,5-Dichloro-6-hydroxy-2-pyridyl) thiopropanoic acid. CONCLUSION: Henceforth, this TCP hapten would be used in developing immunoassay studies for the detection and quantitation of urinary TCP of farmers.




  1. Zhang, W., Tang, Y., Du, D., Smith, J., Timchalk, C., Liu, D., & Lin, Y. (2013). Direct analysis of trichloropyridinol in human saliva using an Au nanoparticles-based immunochromatographic test strip for biomonitoring of exposure to chlorpyrifos. Talanta, 114, 261-267.
  2. Mauriz, E., Calle, A., Manclus, J. J., Montoya, A., & Lechuga, L. M. (2007). On-line determination of 3, 5, 6-trichloro-2-pyridinol in human urine samples by surface plasmon resonance immunosensing. Analytical and Bioanalytical Chemistry, 387(8), 2757-2765.
  3. Manclús, J. J., Primo, J., & Montoya, A. (1994). Development of a chlorpyrifos immunoassay using antibodies obtained from a simple hapten design. Journal of Agricultural and Food Chemistry, 42(6), 1257-1260.
  4. Manclús, J. J. & Montoya, A. (1995). Development of immunoassays for the analysis of chlorpyrifos and its metabolite 3,5,7-trichloro-2-pyridinol in the aquatic environment. Analytica Chimica Acta, 311, 341-348.
  5. Kim, Y. A., Lee, E. H., Kim, K. O., Lee, Y. T., Hammock, B. D., & Lee, H. S. (2011). Competitive immunochromatographic assay for the detection of the organophosphorus pesticide chlorpyrifos. Analytica Chimica Acta, 693(1), 106-113.
  6. Gorton, L. (Ed.). (2005). Biosensors and modern biospecific analytical techniques (p. 331), Elsevier B. V., The Netherlands.
  7. Chuang, J. C., Van Emon, J. M., Reed, A. W., & Junod, N. (2004). Comparison of immunoassay and gas chromatography/mass spectrometry methods for measuring 3, 5, 6-trichloro-2-pyridinol in multiple sample media. Analytica Chimica Acta, 517(1), 177-185.


연구 과제번호 : 급만성 농약노출 예측을 위한 생체 바이오마커 개발