|Drug Class Name
||Beta-agonists, Clenbuterol, Salbutamol and Cimaterol|
||ELISA Screening for Beta-Agonist Residues in Animal Retinal Tissue|
|Scope and Application
||This ELISA method is suitable for screening the presence of clenbuterol in bovine, ovine, porcine, and caprine retinal tissue as well as salbutamol and cimaterol in bovine and porcine retinal tissue.
||For retinal tissue, the eyelid is carefully incised from a thawed sample. The eyeball is then everted and the choroid/PRE layer scrapped into a petri dish prior to mincing into fine pieces with a razor. Minced bovine, ovine, caprine, or porcine retinal
tissue sample is then mashed in centrifuge tissue after adding an extraction buffer (potassium phosphate monobasic and sodium phosphate dibasic at pH 6.8). This is then centrifuged and aliquots of the supernatant added into duplicate wells of the Generic Bronchodilator
ELISA plate. Diluted Terbutalene-HRP Conjugate Solution (HRP) and EIA buffers are added to each well and the solution gently mixed and incubated at room temperature before washing with diluted washing buffer. K-Blue substrate is added to each well with the
reaction proceeding for 15 minutes with intermittent gentle shaking. The plate reader is set at 650 nm and the plate read typically after 30 minutes.
|Applicable Concentration Range
||Clenbuterol may be detected in bovine, ovine, porcine, and caprine retinal tissue at ≥ 3 µg kg -1, salbutamol (at ≥ 3 µg kg -1) and cimaterol (at ≥ 6 µg kg -1) may also be detected in bovine and porcine retinal tissue using this method.
||See attached SOP|