Aminoglycoside Antibiotics in Feed Apramycin, Gentamicin, Neomycin

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Courtesy of Pickering Laboratories, Inc.

Aminoglycoside antibiotics are among those commonly used in animal feeds to manage intestinal microorganisms. The beneficial effects include improved growth and generally healthier animal populations. Use of antibiotics creates a demand for analytical procedures to verify concentrations in pre-mixes and feeds and in some instances for residue analysis in animal products. This note describes a simple, robust analytical method for the family of Aminoglycoside antibiotics in feeds and animal products. The sample is homogenized with a generic extraction solution and the crude soluble portion is directly injected into an HPLC ion-exchange column. The column effluent is then mixed with an OPA/Thiofluor reagent which forms highly fluorescent derivatives with the primary amine moieties of the antibiotics.

Extraction Procedure:

Take one part feed:10 parts Extraction solution (w/v) Catalog No. 1700-1118 and homogenize for five minutes. Centrifuge for 10 minutes. Three layers will form: the pellet, a supernatant emulsion and a soft layer of floating fat. Carefully lift the floating fatty layer with a spatula and discard. Transfer the emulsion to a sealable vial. Coagulate the emulsion by placing the vial in a boiling water bath for 15 minutes. Centrifuge for 10 minutes. The clear supernate is filtered (0.45um Nylon) and placed in an autosampler vial.

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