| 1 ObjectiveTo identify the chemical components in the blood and Cerebro-Spinal Fluid,an ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method was developed based on chemical constituent identification and fingerprint analysis of the blood and cerebro-spinal fluid samples from Yindan xinnaotong Capsule following oral administration to rat. In addition, to investigate the pharmacokinetics and tissue distribution of Yindanxinnaotong Capsule an ultra high performance liquid chromatography with triple-quadrupole mass spectrometery(UPLC-QQQ-MS) method was developed and validated for simultaneous quantification of seven compounds plasma, brain and cerebro-spinal fluid in rat. The results above are as follows:2 Methods and Results2.1 Identification of the blood and cerebro-spinal fluid samples from Yindan xinnaotong Capsule following oral administration to rat by ultra high performanceliquid chromatography with quadrupole time-of-fli ght mass spectrometry(UPLC-Q-TOF-MS) fingerprintingIn this study, ultra high performance liquid chromatography coupled with time-of-flight mass spectrometry was used to identify the chemical components of the blood and cerebro-spinal fluid(CSF) samples from Yindan xinnaotong Capsule following oral administration to rat. Approximately 18 components were tentatively identified in blood, including ginkgolide, phenolic acid, diterpenoid tanshinones, and ginsenoside; And 8 components were tentatively identified in CSF, which are both come from blood.2.2 Pharmacokinetics of Yindan xinnaotong Capsule following oral administration to ratDue to the similar structures, GA, GB, GC, BB, QCT, KMF, ISR, TSⅡA andSAB are absorbed and eliminated with similar rates in vivo, resulting in parallel pharmacokinetic parameters and concentration–time curves. The time period required to reach the maximum plasma concentration(Tmax) was 0.75 h for GA, 1.00 h for GB, 1.50 h for GC, 0.75 h for BB, 0.33 h for QCT, 0.50 h for KMF, 0.33 h for ISR, 0.25 h for TSⅡA and 0.75 h for SAB. Additionally, flavonols were eliminated faster than terpene lactones. The Cmax values and the mean AUC0-t for GA, GB, GC, BB,QCT,ISR,KMF, TSⅡA, SAB were 76.31 ng·m L-1 and 490.92 ng·m L-1·h-1, 76.54 ng·m L-1 and 610.18 ng·m L-1·h-1, 35.35 ng·m L-1 and 281.80 ng·m L-1·h-1, 48.70 ng·m L-1 and 365.97 ng·m L-1·h-1, 45.02 ng·m L-1 and 410.34 ng·m L-1·h-1, 27.85 ng·m L-1 and 319.17 ng·m L-1·h-1, 49.90 ng·m L-1 and 401.33 ng·m L-1·h-1, 38.34 ng·m L-1 and 167.20 ng·m L-1·h-1, 32.00 ng·m L-1 and 118.17 ng·m L-1·h-1,respectively. The T1/2 values ranging from 2.69 h to 12.17 h, suggesting the nine bioactive compounds were slowly eliminated in rat.Additionally, the flavonols showed double peaks in curves of mean plasma concentration. This is a phenomenon that has been reported previously which indicates that these components might have enterohepatic recirculation.The double-peaks suggested that the enterohepatic recirculation, as well as inter-transformation among the compounds, might have occurred. One possible explanation for this phenomenon is that some of the other compounds with similar structures might have transform to these compounds. Since drug absorption is a complex process, more detailed adsorption studies are needed to ascertain the mechanism of the double-peak phenomenon.2.3 Distribution in the brain and CSF of Yindan xinnaotong Capsule following oral administration to ratThe distribution of GA, GB, GC, BB and TSⅡA was investigated in rat brain and CSF following i.g. of the YXC(4.8 g·kg-1). Results indicated that all analytes underwent a rapid and wide distribution into tissues within the time course examined, with no long-term accumulation in tissues observed. the highest brain concentration of GA was observed in the 1 h(18.98 ng·mg-1),in CSF was 1 h(3.43 ng·mg-1).For GB,the highest brain concentration of was observed in the 1 h(23.67 ng·mg-1),in CSF was 1 h(9.21 ng·mg-1). For GC, the highest brain concentration of was observed in the 1 h(21.43 ng·mg-1), in CSF was 1 h(6.64 ng·mg-1). For BB, the highest brain concentration of was observed in the 1 h(23.11 ng·mg-1), in CSF was 1 h(7.22 ng·mg-1). For TSⅡA, the highest brain concentration was observed in the 1 h(15.10 ng·mg-1), and we can’t find it in CSF. As we know, YXC has curative effect in both cardiovascular and cerebrovascular system diseases, our study showed that the concentrations of GA, GB, GC, BB, and TSⅡA were low in brain. This result may be related to the presence of the blood brain barrier. More detailed absorption studies are needed to understand the curative effect of YXC in cerebrovascular system diseases.3. Conclusion 18 components were tentatively identified in blood, including ginkgolide, phenolic acid, diterpenoid tanshinones, and ginsenoside; and 8 components were tentatively identified in CSF, which are both come from blood. The pharmacokinetics result of Yindanxinnaotong Capsule indicated that YXC has a good absorption and slowly eliminated in rat.The concentration in brain or CSF of GA, GB, GC, BB, and TS ⅡA were low,it may be related with the blood brain barrier. |
PDF Full Text Request