||Other Veterinary Drugs|
|Drug Class Name
||Gentamicin, An aminogylcoside|
||Gentamicin in Porcine Tissue by HPLC|
|Scope and Application
||This method is suitable for screening and quantifying gentamicin in porcine tissue.
||Porcine tissue homogenized in 1 N H2 SO4 is extracted by centrifugation and the supernatant transferred to a beaker. The pellet is re-centrifuged in water and the two supernatants combined. The resulting solution now adjusted to a pH of 4.0 is further
centrifuged and the extract cleaned-up by column chromatography on an IRC50 ammonia phase column onto which a glass wool plug has been placed. After a series of cleanup steps, the analyte is eluted from the column using NH4OH. This eluate is then subjected
to purification byliquid-liquid solvent extraction. Here chloroform is added to the column eluate and the mixture centrifuged after mixing on a vortex. The chloroform is then discarded and the remaining aqueous layer subjected to cleanup on a Sep-Pak chromatographic
cartridge. After another series of cleanup steps, the analyte is eluted using a Sep-Pak elution solvent. The aqueous solution is diluted with water before adding netilmicin. The eluate is now ready for HPLC-fluorometric analysis with post column derivatization
using o-phthalaldehyde. A CPRP-1 analytical column is used for separation.|
|Applicable Concentration Range
||Gentamicin may be detected/quantified in porcine tissue at the concentration level ≥ 0.4 µg kg -1.
||See attached SOP|