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J Sep Sci. 2018 Nov;41(22):4115-4121. doi: 10.1002/jssc.201800683. Epub 2018 Oct 3.

Selective and sensitive determination of tetracyclines by HPLC with chemiluminescence detection based on a cerium(IV)-methoxylated cypridina luciferin analogue system.

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Jiangxi Province Key Laboratory of Preventive Medicine, School of Public Health, Nanchang University, Nanchang, P. R. China.
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University, Jinan, P. R. China.


A novel approach for the simultaneous and selective analysis of chlortetracycline, doxycycline, 4-epichlortetracycline, oxytetracycline, metacycline, and tetracycline by high-performance liquid chromatography coupled with sensitive post-column chemiluminescence reaction was set up. Six tetracyclines were separated through a simple and effective isocratic procedure using a mixture of acetonitrile and 3.5 mM methanesulfonic acid. After that, intensive chemiluminescence signals were produced by the reaction between cerium(IV) and methoxylated cypridina luciferin analogues system in the presence of tetracyclines. Under the appropriate experimental conditions, the developed method was evaluated by testing of linearity, limits of detection, precision, and accuracy. The limit of detection was from 2 to 10 nM and relative standard deviation of intraday precision was below 9%. The developed high-performance liquid chromatography with chemiluminescence method has been performed for the with trace analysis of tetracycline antibiotic residues in milk samples with satisfactory results.


antibiotics; cerium; chemiluminescence detection; high-performance liquid chromatography; tetracyclines

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