Veterinary Drugs Residue Methods


Category Steroids
Drug Class Name Anabolic Steroid
Method Title Confirmation of alpha and Beta -Trenbolone by GC/MS/MS
Method Date 2005/01/03
Method Type Confirmatory, Quantitative
Scope and Application This method can be used to confirm the presence of alpha-trenbolone and Beta-trenbolone in bovine liver and muscle, respectively.
Method Summary Homogenized liver/muscle tissue in 0.04 M sodium acetate buffer is digested with beta glucuronidase and extracted in acetonitrile by centrifugation. The supernatant is mixed with sodium hydroxide, hexane and dichloromethane prior to shaking and subsequent centrifugation. Collect the middle layer (after repeated extraction) and mix with water. Centrifuge again and then discard the bottom aqueous layer. Evaporate remaining solution to dryness and dissolve the residue in water before cleaning up using C18 cartridges. The analyte is subsequently eluted with 80 to 20 (methanol, water) from the cartridge. The eluate is dissolved in toluene, centrifuged and stored at -10 degrees celcius overnight before further cleanup using silica cartridges. here the analyte is eluted with 20 to 80 (acetone, toluene). This is dried and the residue dissolved in ethyl acetate and then mixed with BSTFA, TMSI derivatization agent prior to GCMSMS (Ion-trap) analysis. A GC analytical column DB5 ms, 30m, 0.25 mm i.d., 0.25 micrometer film thickness is used.
Applicable Concentration Range Alpha trenbolone and Beta trenbolone may be detected in bovine liver and muscle at the concentration level at least 5 microgram per kilogram
QC Requirements See attached SOP
Method Source USDA-FSIS
Method SOP PDF icon SOP