Veterinary Drugs Residue Methods


Category Beta-agonists
Drug Class Name Beta agonists including clenbuterol, salbutamol, cimaterol, zilpaterol, and ractopamine
Method Title Screening and Confirmation of Beta-Agonists by HPLC/MS/MS
Method Date 2012/02/07
Method Type Confirmatory, Quantitative
Scope and Application This method can be used to screen and or confirm a number of beta agonists in bovine retinal tissue (except zilpaterol) and bovine, porcine, ovine, and caprine liver as well as bovine and porcine muscle.
Method Summary The eyelid is carefully incised from a thawed sample while the humors are still frozen. The eyeball is then everted and the choroid/PRE layer scrapped into a petri dish prior to mincing into fine pieces with a razor. The samples are then extracted by centrifugation after adding acetonitrile, isopropanol, sodium chloride, sodium sulphate and magnesium sulphate. An aliquot of the extract is evaporated to dryness and then dissolved in water prior to filtration into HPLC vials using 0.45 micrometer nylon filter. Alternatively, a combination of a mini-uniprep filter vial and 0.45 micrometer nylon filter may also be used. Homogenized liver or muscle samples (thawed where necessary) are also prepared as above. Sample analysis is done by LC/MS/MS in positive ESI, MRM mode. A C-18 Thermo Hypersil BetaMax Base column is used so separate analytes
Applicable Concentration Range Clenbuterol, salbutamol, cimaterol and ractopmanine HCl (all at ≥ 3 µg kg-1) may be screened in bovine, porcine, ovine, and caprine liver plus bovine and porcine muscle. While zilpaterol may be screened (≥ 6 µg kg-1). Screening and or confirmation of all (except zilpaterol) is possible in bovine retinal tissue.
QC Requirements See attached SOP
Method Source USDA-FSIS
Method SOP PDF icon SOP 
Citation USDA-FSIS Chemistry Laboratory Guidebook