Abstract: HPLC was applied to the determination of residual amounts of clenbuterol in pork and serum. The sample was extracted with a mixture of methanol and 0.01 mol·L-1 hydrochloric acid (68+32) and after purification with ethylenediamine-N-propylsilane, the sample solution was separated on Zorbax Eclipse C18 chromatographic column with a mixture of methanol and sodium dihydrogen phosphate solution (68+32) as mobile phase. UV detection was made at the wavelength of 270 nm. Linearity range of clenbuterol was between 0.01-5.00 mg·L-1, with detection limit (3S/N) of 1.8 μg·kg-1. Values of recovery obtained by standard addition method were in the range of 87.4%-99.2% , with RSDs (n=5) in the range of 2.1%-2.8%.