| Category |
Other Veterinary Drugs |
| Drug Class Name |
Gentamicin: An aminogylcoside |
| Method Title |
Gentamicin in Porcine Tissue by HPLC |
| Method Date |
1991/07/01 |
| Method Type |
Screening, Quantitative |
| Scope and Application |
This method is suitable for screening and quantifying gentamicin in porcine tissue. |
| Method Summary |
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 IRC-50 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. |
| QC Requirements |
See attached SOP |
| Method Source |
USDA-FSIS |
| Method SOP |
 SOP |
| Citation |
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