Publication: A Fast and Accurate Method for the Quantification of Doxycycline in Goat Plasma and Milk by HPLC Using a Fluorescence Detector
Authors
Martínez, José ; Hernandis, Verónica ; Badillo, Elena ; Escudero, Elisa ; Yuste Pérez, María Teresa ; Galecio, Juan Sebastián ; Marín, Pedro
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Publisher
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DOI
http://dx.doi.org/10.2478/acve-2024-0035
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info:eu-repo/semantics/article
Description
© 2024 The authors
This document is the published version of a published work that appeared in final form in Acta Veterinaria-Beogra. .
This document is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0
To access the final edited and published work see:
http://dx.doi.org/10.2478/acve-2024-0035
Abstract
Doxycycline is an antimicrobial agent used in veterinary medicine to treat a variety of
bacterial infections. To date, no analytical technique utilising HPLC with fl uorescence
detection has been documented for the quantifi cation of doxycycline concentrations in
goat plasma or milk. Consequently, the objective of the present study was to propose
a rapid HPLC assay with fl uorescence detection for the quantifi cation of doxycycline
in the aforementioned samples, thereby facilitating the conduct of pharmacokinetic
studies and the detection of residues in diverse goat tissues. Proteins were precipitated
with methanol and trifl uoroacetic acid in a single step. Doxycycline was separated on
a XBRIDGE C18 column using an isocratic method. Sample volume injected into the
HPLC system was 50 µl. Fluorescence detection was conducted with an excitation
wavelength of 380 nm and an emission wavelength of 520 nm. The retention times
of doxycycline and danofl oxacin (internal standard) were determined to be 8.0 and 5.5
minutes, respectively. The calibration curves for plasma and milk exhibited linearity
over the concentration range of 0.1 to 2 μg/mL. The limit of detection was 0.065 μg/
mL, while the limit of quantifi cation was 0.1 μg/mL in both matrices. The accuracy and
precision of the method were consistently within the limits of 10.9% for plasma and
10.5% for milk. The fi ndings of this study may be employed in the quantifi cation of
doxycycline in goat plasma and milk, thus facilitating the conduct of pharmacokinetic
studies.
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Citation
Acta Veterinaria-Beograd 2024, 74 (4), 511-522
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