Molecules and Cells

  • 제21권2호
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  • Pages.308-313
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  • 2006
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  • 1016-8478(pISSN)
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  • 0219-1032(eISSN)
한국분자세포생물학회 (Korean Society for Molecular and Cellular Biology)
권호 표지

Detection of Abnormally High Amygdalin Content in Food by an Enzyme Immunoassay

  • Cho, A-Yeon (Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine) ;
  • Yi, Kye Sook (Transplantation Research Institute, Seoul National University College of Medicine) ;
  • Rhim, Jung-Hyo (Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine) ;
  • Kim, Kyu-Il (Department of Chemisty, Chonbuk National University) ;
  • Park, Jae-Young (Department of Food and Nutritional Science, Ewha Womans University) ;
  • Keum, Eun-Hee (Department of Food and Nutritional Science, Ewha Womans University) ;
  • Chung, Junho (Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine) ;
  • Oh, Sangsuk (Department of Food and Nutritional Science, Ewha Womans University)
  • 투고 : 2005.12.15
  • 심사 : 2006.03.07
  • 발행 : 2006.04.30
⟨ 이전 논문 다음 논문 ⟩

초록

Amygdalin is a cyanogenic glycoside compound which is commonly found in the pits of many fruits and raw nuts. Although amygdalin itself is not toxic, it can release cyanide (CN) after hydrolysis when the pits and nuts are crushed, moistened and incubated, possibly within the gastrointestinal tract. CN reversibly inhibits cellular oxidizing enzymes and cyanide poisoning generates a range of clinical symptoms. As some pits and nuts may contain unusually high levels of amygdalin such that there is a sufficient amount to induce critical CN poisoning in humans, the detection of abnormal content of amygdalin in those pits and nuts can be a life-saving measure. Although there are various methods to detect amygdalin in food extracts, an enzyme immunoassay has not been developed for this purpose. In this study we immunized New Zealand White rabbits with an amygdalin-KLH (keyhole limpet hemocyanin) conjugate and succeeded in raising anti-sera reactive to amygdalin, proving that amygdalin can behave as a hapten in rabbits. Using this polyclonal antibody, we developed a competition enzyme immunoassay for determination of amygdalin concentration in aqueous solutions. This technique was able to effectively detect abnormally high amygdalin content in various seeds and nuts. In conclusion, we proved that enzyme immunoassay can be used to determine the amount of amygdalin in food extracts, which will allow automated analysis with high throughput.

키워드

과제정보

연구 과제 주관 기관 : Agricultral Research Promotion Center

참고문헌

  1. Backofen, U., Matysik, F. M., and Werner, G. (1996) Determination of cyanide in microsamples by means of capillary flow injection analysis with amperometric detection. J. Anal. Chem. 356, 271-273
  2. Campa, C., Schitt-Kopplin, P. H., Catldi, T. R. I., Bufo, S. A., Freitaq, D., et al. (2000) Analysis of cyanogenic glycosides by micellar capillary electrophoresis. J. Chromatogr. B. Biomed. Sci. Appl. 739, 95-100 https://doi.org/10.1016/S0378-4347(99)00375-8
  3. Chassagne, D., Crouzet, J. C., Bayonove, C. L., and Baumes, R. L. (1996) Identification and quantification of passion fruit cyanogenic glycosides. J. Agric. Food. Chem. 4444, 3817-3820
  4. Curtis, A. J., Grayless, C. C., and Fall, R. (2002) Simultaneous determination of cyanide and carbonyls in cyanogenic plants by gas chromatography-electron capture/photoionization detection. Analyst 127, 1446-1449 https://doi.org/10.1039/b205378k
  5. Gomez, E., Burgos, L., Soriano, C., and Marin, J. (1998) Amygdalin content in the seeds of several apricot cultivars. J. Sci. Food Agric. 77, 184-186 https://doi.org/10.1002/(SICI)1097-0010(199806)77:2<184::AID-JSFA22>3.0.CO;2-H
  6. Hughes, C., Lehner, F., Dirkolu, L., Harkins, D., Boyles, J., et al. (2003) A simple and highly sensitive spectrophotometric method for the determination of cyanides in equine blood. Toxicol. Mech. Meth. 13, 129-138 https://doi.org/10.1080/15376510309847
  7. Hwang, E. Y., Lee, J. H., Lee, Y. M., and Hong, S. P. (2002) Reverse-phase HPLC separation of D-amygdalin and neoamygdalin and optimum conditions for inhibition of racemization of amygdalin. Chem. Pharm. Bull. 50, 1373-1375 https://doi.org/10.1248/cpb.50.1373
  8. Keusgen, M., Milka, P., and Krest, I. (2001) Cyanides from bacterial sources and its potential for the construction of biosensors. BIOSENSOR SYMPOSIUM 2001
  9. Koo, J. Y., Hwang, E. Y., Cho, S., Lee, J. H., Lee, Y. M., et al. (2005) Quantitative determination of amygdalin epimers from armeniacae semen by liquid chromatography. J. Chromato. B. 814, 69-73 https://doi.org/10.1016/j.jchromb.2004.10.019
  10. Marple, R. L., Li, X., and LaCourse, W. R. (2004) Pulsed electrochemical detection of aryl- and alkylglycosides following reverse-phase liquid chromatography. J. Liquid Chromato. Rel. Tech. 40, 1695-1710
  11. Moraes, L. A. B., Eberlin, M. N., Cagnon, J. R., and Urbano, L. H. (2000) A new method for the selective quantitation of cyanogenic glycosides by membrane introduction mass spectrometry. Analyst 125, 1529-1531 https://doi.org/10.1039/b004300l
  12. Nout, M. J. R., Tuncel, G., and Brimer, L. (1995) Microbial degradation of amygdalin of bitter apricot seeds. Int. J. Food Microbiol. 24, 407-412 https://doi.org/10.1016/0168-1605(94)00115-M
  13. Oomah, B. D., Mazza, G., and Kenaschuk, E. O. (1992) Cyanogenic compounds in flaxseed. J. Agric. Food Chem. 40, 1346-1348 https://doi.org/10.1021/jf00020a010
  14. Tatsuma, T., Komori, K., Yeoh, H. H., and Oyama, N. (2000) Disposable test plates with tyrosinase and beta glycosidases for cyanide and cyanogenic glycosides. Analytica. Chemica. Acta 408, 233-240 https://doi.org/10.1016/S0003-2670(99)00744-8
  15. Zlosnik, J. E. A. and Williams, H. D. (2004) Methods for assaying cyanide in bacterial culture supernatant. Lett. Appl. Microbiol. 38, 360-365 https://doi.org/10.1111/j.1472-765X.2004.01489.x