Liquid chromatographic determination of oxytetracycline in bovine plasma by double-phase extraction.
A liquid chromatographic method for the determination of oxytetracycline in bovine plasma was developed. Contrary to most current tetracycline assays involving solid-phase extraction, oxytetracycline and the internal standard tetracycline were isolated from buffered plasma by double-phase extraction with ethyl acetate-isopropyl alcohol. The latter component was found to be essential to ensure reproducible partitioning of both tetracyclines into the organic layer. Chromatography was conducted on a Lichrosorb RP8 column, compounds were eluted with 0.01 M oxalic acid-methanol-acetonitrile, and detection was at 357 nm. Linearity was observed over the 0-8.5-micrograms/mL range, and the detection limit approximated 5 ng/mL. The extraction yield was 80.4 +/- 3.4% (n = 12), and the coefficients of variation for repetitive within-run and between-run analyses at two concentration levels (0.25 and 2 micrograms/mL) were 1.0-4.3 and 2.4-7.2%, respectively. The method was applied to a pharmacokinetic study of oxytetracycline in three nonlactating cows after a single intramuscular administration of a long-acting preparation.
Nelis
H J
HJ
Laboratoria voor Medische Biochemie en voor Klinische Analyse, Ghent, Belgium.
Vandenbranden
J
J
De Kruif
A
A
Belpaire
F
F
De Leenheer
A P
AP
D000433:1-Propanol; D000085:Acetates; D000818:Animals; D002417:Cattle; D002623:Chemistry Techniques, Analytical / Q000379:methods; D002853:Chromatography, Liquid; D005260:Female; D010118:Oxytetracycline / Q000097:blood*
D000085:Acetates; C007650:ethyl acetate; D000433:1-Propanol; D010118:Oxytetracycline