Validation of a high-performance liquid chromatographic method for the determination of doxycycline in turkey plasma.
A high-performance liquid chromatographic method for the analysis of doxycycline in turkey plasma samples using demeclocycline hydrochloride as the internal standard was developed, optimized and validated. A one-step extraction procedure and an isocratic HPLC method with UV detection were used. No interferences with endogenous compounds or with the anticoagulant were observed. Linear calibration curves (r2 > 0.99) were obtained in water and plasma between 0 and 600 micrograms ml-1. Good recoveries for doxycycline (> 66%) and demeclocycline (> 72%) were seen both in water and in plasma. The coefficient of variation was < 9.86% for within-day reproducibility and < 7.53% for the between-day reproducibility. The deviation between the mean value found and the true value was < 14.5% (accuracy). The limit of detection was 0.1 microgram ml-1 in plasma samples. A good stability of doxycycline was observed in water and in plasma samples after storage for six months at -20 degrees C (recovery > 91%).
Santos
M D
MD
Laboratory of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, University of Ghent, Belgium.
Vermeersch
H
H
Remon
J P
JP
Schelkens
M
M
De Backer
P
P
Ducatelle
R
R
Haesebrouck
F
F
D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004318:Doxycycline / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014422:Turkeys
D000900:Anti-Bacterial Agents; D004318:Doxycycline