- P-ISSN 1225-0163
- E-ISSN 2288-8985
A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in bovine, porcine, chicken, flatfish and japanese eel muscle has been developed and validated. The separation condition for HPLC/UV was optimized with phenyl hexyl (<TEX>$4.6{\times}150mm$</TEX>, <TEX>$5{\mu}m$</TEX>) column with 10 mM monobasic sodium phosphate buffer (pH 2.5)/acetonitrile (50/50, v/v) as the mobile phase at a flow rate of 1.0 mL/min and detection wavelength was set at 254 nm. Residues were extracted from tissue by blending with methanol and lipid materials were removed with n-hexane. Then, the methanol extract was evaporated to dryness under a nitrogen stream, reconstituted in the mobile phase. Aliquot of the organic extract was decanted and filtered through <TEX>$0.45{\mu}m$</TEX> syringe filter. The <TEX>$20{\mu}L$</TEX> of the resulting solution was injected into the HPLC system. The calibration ranges were <TEX>$0.5{\sim}5{\mu}g/g$</TEX> and calibration curves were linear with coefficients of correlation better than 0.95. The limits of quantification were <TEX>$0.5{\mu}g/g$</TEX> for all muscles. The recoveries of bovine, porcine, chicken, flatfish and japaneseel muscles were 99.8%, 102.4%, 91.0%, 104.0% and 93.0%, respectively. The procedures were validated according to the CODEX guideline, determining specificity, linearity, accuracy, precision, quantitation limit and recovery.
The Merck Veterinary Manual ,
The Greenbook ,
M. A. Montecalvo, (1995) J. Chromatogr. B,,
Part 556.460, (1992) Code of Federal Regulations,
(1993) Codex Guidelines for the Establishment of a RegulatoryProgramme for Control of Verterinary Drug Residuesin Foods,
(1998) Validation Guidelines,
V. B. Reeves, (1995) Antimicrob,
H. J. Kim, (1998) J. Vet. Serv,
A. Zarghi, (2005) J. Pharm. Biomed. Anal,