Enzyme-Linked Immunosorbent Assay of Ampicillin in Milk

  • Zh. V. Samsonova
  • O. S. Shchelokova
  • N. L. Ivanova
  • M. Yu. Rubtsova
  • A. M. Egorov


An indirect immunoassay for quantitative determination of ampicillin (range, 10–1000 ng/ml) in buffer or milk has been developed. Polyclonal antibodies were obtained against ampicillin conjugated with bovine serum albumin; the conjugate was synthesized by direct condensation using carbodiimide. The antibodies were specific for ampicillin and exhibited low cross-reactivity to other penicillins (azlocillin, 17%; penicillin G, 10%; piperacillin, 5%; and carbenicillin, 4%). Matrix effects were minimized by combining the use of a casein-supplemented buffer (content of casein, 1%) with sample dilution. Limit of detection for ampicillin in milk (diluted tenfold) was equal to 5.0 ng/ml (which corresponded to 50 ng/ml of the original sample).


Albumin Bovine Serum Albumin Serum Albumin Penicillin Ampicillin 
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  1. 1.
    Gigienicheskie trebovaniya bezopasnosti i pishchevoi tsennosti pishchevykh produktov. SanPiN (Hygienic Requirements for Safety and Food Value of Food Products), Moscow: Minzdrav Rossii, 2002, p. 168.Google Scholar
  2. 2.
    Egorov, N.S., Osnovy ucheniya ob antibiotikakh (Fundamentals of the Theory of Antibiotics), Moscow: Mosk. Gos. Univ., 1994.Google Scholar
  3. 3.
    Council Regulation (EEC) no. 2377/90, Off. J. Eur. Commun., 1990, vol. 224, pp. 1–8.Google Scholar
  4. 4.
    Riediker, S., Diserens, J.M., and Stadler, R.H., J. Agric. Food Chem., 2001, vol. 49, no.9, pp. 4171–4176.CrossRefPubMedGoogle Scholar
  5. 5.
    Althaus, R.L., Molina, M.P., Rodriguez, M., and Fernandez, N., J. Food Prot., 2001, vol. 64, no.11, pp. 1844–1847.PubMedGoogle Scholar
  6. 6.
    Sorensen, L.K., Rasmussen, B.M., Boison, J.O., and Keng, L., J. Chromatog. Ser. B, 1997, vol. 694, no.2, pp. 383–391.Google Scholar
  7. 7.
    Gazzaz, S.S., Rasco, B.A., and Dong, F.M., Crit. Rev. Food Sci. Nutr., 1992, vol. 32, no.3, pp. 197–229.PubMedCrossRefGoogle Scholar
  8. 8.
    Martlbauer, E., Usleber, E., Scheneider, E., and Dietrich, R., Analyst, 1994, vol. 119, no.12, pp. 2543–2548.PubMedGoogle Scholar
  9. 9.
    Jackman, R., Michell, S.J., Dyer, S.D., and Chesham, J., Food Agric. Immunol., 1991, vol. 3, no.1, pp. 3–12.Google Scholar
  10. 10.
    Usleber, E., Lorber, M., Straka, M., Terplan, G., and Martlbauer, E., Analyst, 1994, vol. 119, no.12, pp. 2765–2768.CrossRefPubMedGoogle Scholar
  11. 11.
    Rohner, P., Schallibaum, M., and Nicolet, J., J. Food Prot., 1995, vol. 48, no.1, pp. 59–62.Google Scholar
  12. 12.
    Sternesjo, A. and Johnsson, G., J. Food Prot., 1998, vol. 61, no.7, pp. 808–811.PubMedGoogle Scholar
  13. 13.
    Huth, S.P., Warholic, P.S., Devou, J.M., Chaney, L.K., and Clark, G.H., J. AOAC Int., 2002, vol. 85, no.2, pp. 355–364.PubMedGoogle Scholar
  14. 14.
    Nakamura, H. and Karube, I., Anal. Bioanal. Chem., 2003, vol. 377, no.3, pp. 446–468.CrossRefPubMedGoogle Scholar
  15. 15.
    Gaudin, V., Fontaine, J., and Maris, P., Anal. Chim. Acta, 2001, vol. 436, no.2, pp. 191–198.CrossRefGoogle Scholar
  16. 16.
    Gustavsson, E., Bjurling, P., and Sternesjo, A., Anal. Chim. Acta, 2002, vol. 468, no.2, pp. 153–159.Google Scholar
  17. 17.
    Setford, S.J., Van Es, R.M., Blankwater, Y.J., and Kroger, S., Anal. Chim. Acta, 1999, vol. 398, no.1, pp. 13–22.CrossRefGoogle Scholar
  18. 18.
    De Haan, P., de Jonge, A.J., Verbrugge, T., and Boorsma, D.M., Int. Arch. Allergy Appl. Immunol., 1985, vol. 76, no.1, pp. 42–46.PubMedGoogle Scholar
  19. 19.
    Nagakura, N., Souma, S., Shimizu, T., and Yanagihara, Y., J. Antimicrob. Chemother., 1991, vol. 28, no.3, pp. 357–368.PubMedGoogle Scholar
  20. 20.
    Mayorga, C., Obispo, T., Jimeno, L., and Blanca, M., Moscoso Del Prado J., Carreira J., Garcia J.J., Juarez C, Toxicology, 1995, vol. 97, no.2, pp. 225–234.PubMedGoogle Scholar
  21. 21.
    Usleber, E., Litz, S., and Martlbauer, E., Food Agric. Immunol., 1998, vol. 10, no.4, pp. 317–324.CrossRefGoogle Scholar
  22. 22.
    Dietrich, R., Usleber, E., and Martlbauer, E., Analyst, 1998, vol. 123, no.12, pp. 2749–2754.CrossRefPubMedGoogle Scholar
  23. 23.
    Cliquet, P., Cox, E., Van Dorpe, C., Schacht, E., and Goddeeris, B.M., J. Agric. Food Chem., 2001, vol. 49, no.7, pp. 3349–3355.CrossRefPubMedGoogle Scholar
  24. 24.
    Kolosova, A.Yu., Samsonova, Zh.V., Egorov, A.M., Sheveleva, S.A., Orlova, N.G., Kiseleva, T.V., Khotimchenko, S.A., and Tutel'yan, V.A., Vopr. Pitan., 1999, vol. 68, no.1, pp. 23–27.PubMedGoogle Scholar

Copyright information

© MAIK "Nauka/Interperiodica" 2005

Authors and Affiliations

  • Zh. V. Samsonova
    • 1
  • O. S. Shchelokova
    • 1
  • N. L. Ivanova
    • 1
  • M. Yu. Rubtsova
    • 1
  • A. M. Egorov
    • 1
  1. 1.Faculty of ChemistryMoscow State UniversityRussia

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