Detection of clenbuterol in bovine retinal tissue by highperformance liquid chromatography with electrochemical detection

A method for the detection of the P-agonist drug clenbuterol in bovine retinal tissue has been developed. The extraction procedure involves sonication and centrifugation. followed by the addition of ethylenediaminetetraacetic acid (EDTA) to the supernatant. The pH of the supernatant is then brought to 12.2, which is then allowed to sit for 2 h. This is followed by a diethylether extraction. The diethylether extract is dried under nitrogen and the residue is dissolved in 1% formic acid. The quantitation of clenbuterol was accomplished by high-performance liquid chromatography with electrochemical detection. The electrochemical detector was an amperometric detector. The detector was set in the pulsed mode. The oxidizing potential of a carbon electrode was 1.3 V vs. a Ag/AgCl reference electrode and was pulsed to a reduction potential of -2.0 V vs. a Ag/AgCl reference electrode. The limit of detection for this method was 5 ng/ml of clenbuterol (S/N=3). Typical spiked recoveries are 75%.