Comprehensive validation of a liquid chromatography-tandem mass spectrometry method for the confirmation of chloramphenicol in urine including stability of the glucuronide conjugate and efficiency of deconjugation

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Murielle GaugainE Verdon

Abstract

This paper describes a method to reveal the illegal use of chloramphenicol (CAP) in animals intended for human consumption based on the detection of free CAP and chloramphenicol-glucuronide (CAP-glu) in urine. It details the different steps of the method, including hydrolysis of CAP-glu, extraction and cleanup with molecularly imprinted polymers and detection by LC-MS/MS, as well as the validation design. The efficiency of chloramphenicol release during the hydrolysis step and the stability of CAP-glu in urine samples stored at -20°C were also investigated. These verifications were important to ensure the method's suitability for checking CAP misuse in veterinary medicine. Validation results were fully compliant with the qualitative and quantitative criteria required by European regulations. Intraday relative standard deviations were all below 7.5%, while interday relative standard deviations were below 6.9%. Recoveries lay between 93.3 and 104.6%. Purification appears very effective since no matrix effect was demonstrated. CAP-glu was found to be stable for at least 3 months, and the mean recovery following deconjugation was assessed to be 79.4%. The decision limits (CCa) were all found to be lower than 0.1μg/kg.

References

Jan 1, 1994·Drug Metabolism Reviews·G F Bories, J P Cravedi
Jun 19, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Maciej J BoguszMohammed Al-Tufail
Dec 13, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jochen TuerkHeinz-Martin Kuss

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Citations

May 8, 2016·Talanta·Oliver Wahl, Ulrike Holzgrabe
Jan 27, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Tomasz ŚniegockiAndrzej Posyniak

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