Pharmacokinetics of Tanshinone ⅡA in Rats Cerebrospinal Fluid by Microdialysis Combined with UPLC-MS/MS

OBJECTIVE To investigate the pharmacokinetics of tanshinone IIA(TSA) in vivo normal and cerebral ischemia/reperfusion injury(CIRI) rats cerebrospinal fluid by microdialysis combined with UPLC-MS/MS.

METHODS The effects of different perfusion flow rates and TSA concentrations between the relative recovery rate and the relative loss rate were analyzed by incremental and reduction methods. The reduction method was also applied to investigate the stability of the probe recovery rate in 8 h. Cerebral dialysates were collected in normal and middle cerebral artery occlusion/reperfusion rats(MCAO/R) which were injected with TSA internationally. Then content of TSA in samples were quantified by UPLC-MS/MS that was established and verified. Meanwhile, drew the drug time curve and calculated the pharmacokinetic parameters.

RESULTS The relative recovery rate and relative loss rate of the microdialysis probe in vitro were nearby. The increased of perfusion velocity, the decreased of the relative recovery rate as well as the relative loss rate. In addition, both of them were independent of TSA concentration. The relative loss rate in vivo was stable between 1.5 and 8.0 h. The methods of UPLC-MS/MS was excellent for the determination of TSA in rats cerebral dialysate. TSA of cerebral dialysates in normal and middle cerebral artery occlusion/reperfusion model rats was up to the highest at 60 min and 80 min respectively, but the half-life was the same of 69.32 min. Its concentration was significantly different(P<0.05) when the rats were intervened by varied doses of TSA. The higher of TSA, the more peak concentration and area under curve. The mean residence times in all groups were 102.90−170.48 min.

CONCLUSION The established methods of TSA in microdialysis and UPLC-MS/MS are applied to investigate the kinetic in vivo brain feasibly and accurately within treatment time window. The results indicates that TSA is able to cross the blood-brain barrier and enter the brain fast. The pathological state of CIRI will increase the distribution of TSA in the brain. This study provides a medical basis of TSA in the early prevention and treatment of CIRI.