OBJECTIVE: To establish the method for the determination of carteolol hydrochloride concentration in plasma of Beagle dogs, and to study pharmacokinetics of Carteolol hydrochloride orally disintegrating tablets in Beagle dogs. METHODS: The plasma samples were pre-treated by acetonitrile precipitation. HPLC method was adopted. The determination was performed on VP-ODS C18 column with mobile phase consisted of acetonitrile-0.067% disodium hydrogen phosphate solution (13 ∶ 87,V/V) at flow rate of 1.0 mL/min; the detection wavelength was set at 251 nm; the column temperature was 30 ℃; the sample size was 20 μL. Six healthy Beagle dogs were selected and given self-prepared Carteolol hydrochloride orally disintegrating tablets (test preparation) or imported Carteolol hydrochloride tablets (reference preparation), 15 mg/dog by the way of double-preparation double-cycle crossover administration. Venous blood samples were collected at corresponding time points, and plasma concentration of carteolol hydrochloride was determined. The pharmacokinetic parameters of two preparations were calculated by using DAS 2.0 software, and relative bioavailability (F) of test preparation was calculated. The pharmacokinetic parameters of two preparations were compared by SPSS 20.0 software. RESULTS: The linear range of carteolol hydrochloride concentration in plasma was 9.77-1 250 ng/mL (r=0.999 7). The limits of quantitation and detection were 3.6 and 1.2 ng/mL, respectively. The extraction recovery rates were higher than 80%; RSDs of precision and stability tests were all lower than 12%; average RE of accuracy were within -8% to -2%. The main pharmacokinetic parameters of Carteolol hydrochloride orally disintegrating tablets in Beagle dogs were as follows as cmax of (327.82±17.27) ng/mL, tmax of (0.54±0.10) h, AUC0-24 h of (1 136.09±172.69) ng·h/mL, AUC0-∞ of (1 243.52±190.08) ng·h/mL; compared with reference preparation, F0-24 h was 93.03%, F0-∞ was 95.45%. There was statistical significance in lncmax and tmax (P<0.05), but no statistical significance in lnAUC0-24 h (P>0.05) between 2 kinds of preparations. CONCLUSIONS: The established HPLC method is simple, rapid, accurate, precise and sensitive, and can be used for pharmacokinetic study of Carteolol hydrochloride orally disintegrating tablets in Beagle dogs. Compared with imported tablet, absorption rate of Carteolol hydrochloride orally disintegrating tablets in Beagle dogs is faster, without significant difference in absorption degree.