| Objective: To explore a reverse phase high performance liquid chromatography (RP-HPLC) method for the analysis of BBS in biological samples, study the pharmacokinetics (PK) parameters, tissue distribution of BBS for injection in beagle dogs and the elimination pattern in mice in order to provide data to clinic prescription. Methods: ⑴the analysis method of BBS: After protein precipitation with acetonitrile, the BBS in biological samples such as plasma, tissue, urine, bile, excrement reacted with the derivation reagent - benzyl chlorides, and then the product was isolated by solid phase extraction. After that it was separated on the Eclipse XDB C8 (150×4.6mm 5um) column using methanol-water as the mobile phase (flow rate 1.0ml·min-1) and anthracene as the internal standard at 35 ℃ with UV detector at wavelength 232nm. ⑵Investigation of the pharmacokinetics: Following a single iv administration of BBS to beagle dogs in three doses (21.3mg·Kg-1, 10.6mg·Kg-1 and 5.3mg·Kg-1), the changes between the plasma concentration of BBS for injection and the time were observed; ⑶Investigation of tissue distribution: Following a single iv administration of BBS to beagle dogs in dose of 10.6 mg·Kg-1, the tissue distribute characteristic was observed. ⑷Investigation of the elimination: Following a single iv administration of BBS to mice in three doses (72mg·Kg-1, 36mg·Kg-1 and 18mg·Kg-1), the elimination regular of BBS for injection in mice was observed. Results: A two compartment opened model could describe the pharmacokinetics model of BBS in beagle dogs following single iv administration of BBS in three doses respectively: The t1/2(α)of high dose group is 2.10±1.54 min,the t1/2(β) is 119.08±30.19min,the K12 is 0.11±0.10min-1, the K21 is 0.21±0.11min-1, the K10 is 0.0094±0.0013min-1, the Vc is 0.54±0.080 (mg.Kg-1)·(μg·ml-1)-1, the CL(s) is 0.0048±0.0012 mg·Kg-1·min-1·(μg·ml-1)-1, the AUC is 4131.39±804.57(μg·ml-1)·min; The t1/2(α)of mid-dose group is 2.34±0.97min, the t1/2(β)is 113.53±42.00min, the K12 is 0.12±0.034 min-1, the K21 is 0.17±0.079min-1, the K10 is 0.011±0.0050min-1, the Vc is 0.54±0.038(mg.Kg-1)·(μg·ml-1)-1, the CL(s) is 0.00568±0.0026mg·Kg-1·min-1·(μg·ml-1)-1, the AUC is 760.50±708.82(μg·ml-1)·min; The t1/2(α) of low dose group is 1.27±1.21 min, the t1/2 ( β ) is 4.63±16.52min, the K12 is 0.25±0.23min-1, the K21 is 0.29±0.13min-1, the K10 is 0.016±0.038 min-1, the Vc is 0.46±0.22(mg.Kg-1)·(μg·ml-1)-1, the CL(s) is 0.0071±0.0017 mg·Kg-1·min-1·(μg·ml-1)-1, the AUC is 704.49±152.73 (μg·ml-1)·min. There is no obvious difference between the three dose groups. After iv administration of BBS at 5 min, the tissue concentration of BBS reach the peak. And the concentration in kidney was the highest in all tissues, and the concentration in liver was the second. After iv administration of BBS at 90 min, the concentration in each tissue was lower than before, but the concentration in kidney was higher than it in other tissues, the concentration in liver was also the second. Most of the drugs were eliminated from urine; the peak is at 1~3 hour, only small amount of drug was eliminated from bile and excrement. Conclusion: The method for the determination of BBS in biological samples was sensitive, simple, rapid, and accurate. It can be used to determine the concentration of BBS in the biological samples, and study the pharmacokinetics (PK) parameters of it. The pharmacokinetics, tissuedistribution of BBS in beagle dogs as well as the elimination regular of BBS in mice have been studied, which will provide data for clinic prescription, the develop of new dosage form as well as the pharmacological research in other fields.
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