Research On Relationship Between Materical Basement And Clinical Application And Medication Safety Of Pogejiuxin Decoction

ObjectiveResearch on influence on the does of Aconitum carmichaelii Debx,decotion time to the amount of toxic ingredients such as aconitine,hypaconitine,and mesaaconitine from Pogejiuxin Decoction.Alkaloids and other compounds from prescription of Pogejiuxin Decoction were also checked by LC-MS in order to provide material basement to clinical safety of medicine.Methods and Results1)Analysis of Pogejiuxin Decoction in Clinic.2)Research on Dynamic Variation Patterns of Diester Diterpenoids Alkaloids from Aconitum carmichaelii under Different Clinical Decocting Condition.Methods:HPLC analysis was performed on a Technologies Athena C18-WP(4.6 mm×250 mm,5 μm)column with methanol(0.1%triethylamine)-water(70:30)as mobile phase.The flow rate was 1.0 mL min-1,column temperature was fixed at 35 ℃,and the wave length of detection was set at 235 nm.Results:The diester diterpenoid alkaloids were extremely unstable during the decoction of Aconifum carmichaelii,and the content of aconitine showed a tendency of increasing gradually,and then reached the peak value after decocting for 4 hours.The contents of mesaconitine and hypaconitine showed the same decreasing trend.The amount of Hypaconitine was the most after decocting for 0.5 h,and was the least after decocting for 4 h.3)Research on Dynamic Variation Patterns of Diester Diterpenoids Alkaloids in Pogejiuxin Decoction under Different Decocting ConditionHPLC was used to study the dynamic variation of diester diterpenoids alkaloids in Pogejiuxin decoction,and the relationship between the content of diester diterpenoids alkaloids from A.carmichaelii,dosage and decocting time were explored.Methods:The detecting condition was following:the mobile phase system was composed of methanol(contained 0.1%triethylamine)and H2O(V/V=70:30),the flow rate was 1.0 mL-min-1,the temperature of column was 35℃ and the wavelength of detection was 235 nm.Results:The results showed that total contents of diester diterpenoid alkaloids of decoction presented mild changes,and the decreasing sequence as mesaconitine,hypaconitine and aconitine.Moreover,the dissolution rule of diester diterpenoids alkaloids showed as the same as their contents,being increased firstly and then decreased obviously along with time extension,then reached the maximum amount after 2 hours.4)Pogejiuxi Decoctine was analysed by LC-MSMethods:Instruments:LC-MSD-Trap-XCT,mobile phase:Solvent A:H2O-5%formic acid,solvent B:0.0%(Actri).Flow rate:0.400 mL·min-1;ion source:ESI;volumn of injection:1.00 μL;the tempreture of gas:352 ℃;Flow rate of gas:10.06 L·min-1,Draw speed:100 μL min-1,speed of Injection:100 μL min-1,HV capillary:3500V.Results:In Aconitum carmichaelii decoction with decocting time of 0.5 hour,21 compounds were checked by LC-MS.Among of them,4 compounds were determined as ditester aconitines,one compound was single ester aconitines,and 16 compounds were none ester aconitines.In Aconitum carmichaelii decoction with 2 hour decocted,there were also 21 compounds checked by LC-MS,however,their structures of them were different from Aconitum carmichaelii.In Pogejiuxin decoction with decocting time of 0.5 hour,there were 14 compounds checked by LC-MS.3 compounds were ditester aconitines,1 was single ester aconitines,10 were none ester aconitines.In Pogejiuxin decoction with decocting time of 2 hour,there were 22 compounds checked by LC-MS,and they were all aconitines.4 compounds were ditester aconitines,one was single ester aconitines,17 compounds were none ester aconitines.ConclusionThe dosage of A.carmichaelii.in the Pogejiuxin decoction is more than usual dosage in the Chinese Pharmacopoeia,and its range is from 30 g to 200 g.Meanwhile,Pogejiuxin decoctine show significant clinic effect.HPLC and LC-MS were confirmed correctly and quantitatively.Which can be illuminated the dynamic rule of the amount of diester aconitines from A.carmichaelii.When the decoctine time of Pogejiuxin Decoction was decocted 2 hours,not only the amounts of diester aconitines were decreased but also signal ester aconitines,none ester aconitines presented.Above all confirmed toxicity of prescription decreasing and therapeutic effect increasing,and provided material basement to clinical safety of Pogejiuxin Decoction.