| Objective:The purpose of this study was to investigate the change of the constituents of Aconitum alkaloids and total anthraquinones constituents in herb couple of Radix Aconiti Lateralis-Rhei Radix et Rhizomaz(Fuzi-Dahuang) in vitro, and study the Aconitum alkaloids pharmacokinetic differences in the beagle dogs when oral administration of Fuzi decoction and Fuzi-Dahuang decoction, principle of detoxieation and synergism when Fuzi compatibility Dahuang and it’s mechanism of action.Method:1.Choose the Radix Aconiti Lateralis processed products Black Radix Aconiti Lateralis,which often used clinical. Adopting the method of clinical commonly used decoction prepared the Fuzi decoction and Fuzi-Dahuang decoction, HPLC-MS method was used to establish the method of determine four Aconitum alkaloids (hypaconitine(HA), benzoylaco nine(BAC), benzoylhypaconine(BHA), benzoylmesaconine(BMA)) in the Fuzi decoction and Fuzi-Dahuang (2:1,1:1,1:2) decoction; HPLC method was used to establish the method of determine total anthraquinones in the Fuzi decoction and Fuzi-Dahuang (2:1, 1:1,1:2) decoctioa2.Choose healthy beagle dogs as experimental animals, the same dose of Fuzi (equals to 3g crude drug of HSP/kg) was given to beagle dogs for Fuzi decoction and Fuzi-Dahuang(2:1) decoction. HPLC-MS method was developed for simultaneous quantitationat 5,10,20,30.45,60,90,120.180.240.360,480,600,720.1440 min of four Aconitum alkaloids in beagle dog plasma.Using DAS 2.0 software for pharmacokinetic analysis. Using SPSS 19.0 data processing software processing the data, study the Rhizoma onpharmacokinetics of Aconitum alkaloids in beagle dogs plasma ofFuzi-Dahuang(2:1).Results:1.With different proportions of Radix Aconiti Lateralis compatibility Rhubarb, the content levels of HA were increased. But wtih the increase of dosage of Rhubarb,the content gradual declined. While the content levels of BMA, BAC, BHA were decreased and the amount of four aconitum alkaloids decreased after compatibility.The total anthraquinone in Rhubarb showed a downward trend after the compatibility of Radix Aconiti Lateralis and rhubarb, and it decline faster with the increase Radix Aconiti Lateralis.2.After the Rhubarb compatibility Radix Aconiti Lateralis, the content of Afoe-emodin and Rhein in compatibility increased significantly when a ratio of 2:1, and a significant reduction in 1:1,1:2; the content of Emodin, Chrysophanol, physcion after compatibility was significantly decreased; The content of total anthraquinone aftern compatibility was significant declined.3.The validated method was successfully applied to the comparative pharmacokinetic study of HA,BHA, BMA, BAC and in beagle dogs after administration of Fuzi decoction and Fuzi-Dahuang decoction. Four alkaloids and the internal standards obtained good separation, and the matrix effects were between-15% and+ 15%, with no interfering of endogenous substances. Four alkaloids achieved good linear relationship between 0.1-100 ng/mL; RSD values of inter-day precision and intra-day precision were less than 10.747%; extraction recoveries were more than 80% and the RSD was less than 6.972%. It achieved good stability in the condition of storing at room temperature for 8 h, freezing and thawing repeatedly three times and thawing after be stored at-20℃ for 14 days. The method can meet requirements of biological sample analysis and it was feasible.4.Compared with giving Fuzi decoction, when giving the Fuzi-Dahuang decoction. The obsorption of HA、BMA、BAC、BHA were decreased(P<0.05), Cmax of BMA, BAC, BHA were decreased remarkably(P<0.05), Tmax were increased remarkably(P<0.05); MRTo-t of HA, BMA, BAC, BHA were increased, where in the BMA was extended increased (P<0.05), Vd of HA, BMA. BAC were increased.Conclusions:The Compatibility of rhubarb synergistic detoxification mechanism of Radix Aconiti Lateralis may be ralated to lower aconitum alkaloids content in vitro, inhibiting aconitum alkaloids of obsorption and extension of the aconitum alkaloids retention time. |
PDF Full Text Request