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Determination of tetracyclines in meat using two phases freezing extraction method and HPLC-DAD

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Abstract

The protein binding of tetracyclines may cause the common extraction methods are not able to completely remove them from the proteins. To overcome this problem, we introduced a simple, reliable, sensitive, and reproducible method for simultaneous clean-up and preconcentration of 4 tetracyclines in chicken meats based on two phase freezing method.

The tetracyclines were extracted by solid liquid extraction using Britton-Robinson buffer (0.008 mol L−1, pH = 10) from chicken breast, thigh and liver samples. Then simultaneous separation, clean up, protein-drug bonds breaking and preconcentration of tetracyclines from extract solution was performed by two phase freezing method. In this method the acetonitrile was mixed with aqueous extractant solution, the mixture solution was frozen and the organic phase was removed by centrifuge at −20 °C. For separation of tetracyclines a reversed-phase high-performance liquid chromatography was used. In two phase freezing method several parameters such as pH of extract solution, the type of miscible organic solvent, salting out, the ratio of organic/aqueous phases, and extraction cycles were investigated. The recoveries of tetracyclines spiked at three levels of concentration (50, 100, and 150 μg Kg−1) were changed from 76.5 % to 95.5 % with the relative standard deviation values less than 11 %. This method is a reliable tool and could be applied to analyze tetracyclines residues in chicken meats.

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Acknowledgments

We gratefully acknowledge the Vice Chancellor for Research and Technology, Kermanshah University of Medical Sciences for financial support. This work was a section of research proposal in the Vice Chancellor for Research and Technology, Kermanshah University of Medical Sciences for financial support, Kermanshah, Iran.

Compliance with Ethics Requirements

Farhad Ahmadi declares that he has no conflict of interest.

Yasser Shahbazi declares that he has no conflict of interest.

Negin Karami declares that she has no conflict of interest.

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Novel Drug Delivery Research Center, Faculty of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, 67145-1673, Islamic Republic of Iran

    Farhad Ahmadi

  2. Department of Medicinal Chemistry, Faculty of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, 67145-1673, Islamic Republic of Iran

    Farhad Ahmadi

  3. Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Razi University, Kermanshah, 671568-5414, Islamic Republic of Iran

    Yasser Shahbazi & Negin Karami

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  1. Farhad Ahmadi
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  2. Yasser Shahbazi
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Correspondence to Farhad Ahmadi.

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Ahmadi, F., Shahbazi, Y. & Karami, N. Determination of tetracyclines in meat using two phases freezing extraction method and HPLC-DAD. Food Anal. Methods 8, 1883–1891 (2015). https://doi.org/10.1007/s12161-014-0073-7

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