Pharmacokinetics Of Three Alkaloids In Huanglianjiedu Decoction In Diabetic Rats

Huanglianjiedu Decoction is one of the widely applied Chinese medical formulas. It is also the effective formula which can reflect the thought of "the same syndrome for Different Disease".Huanglianjiedu Decoction is a typical one in clearing away the heat-evil and expelling superficial evils.From time immemorial,Huanglianjiedu Decoction has been widely applied in clinical to treat several diseases. So it has a more mature clinical research background. According to the modern pharmacological research ,Huanglianjiedu Decoction can interferes the pathological process of diabetes mellitus in a certain degree effectively.So in this thesis, we pay attention to the three effective alkaloids in Huanglianjiedu Decoction---jatrorrhizine,palmatine,berberine, study the pharmacokinetics of the three alkaloids in rats serum after an intragastrical(i.g.) administration of Huanglianjiedu Decoction to rat ,observe the pharmacokinetic character of the three alkaloids in diabetic rats serum,find out the pharmacokinetics difference of the three alkaloids in two models.The work of this thesis were separated to two parts:document sorting and experimental study.The first part contains the historical background of Huanglianjiedu Decoction,the clinical application of Huanglianjiedu Decoction,the modern pharmacological research of Huanglianjiedu compound,the mechanism study on the treatment of diabetes mellitus by Huanglianjiedu Decoction,the chemical composition of Huanglianjiedu Decoction and the pharmacokinetic study of Huanglianjiedu Decoction .We found Huanglianjiedu Decoction was widely used in clinical which played many roles on disease treatment, such asanti-inflammatory,antimicrobial,antiendotoxin,anti-oxidant,lipid-lowering,hypoglycemi c action,anti-cerebral ischemia, antitumor etc.Among them Huanglianjiedu Decoction had significant curative effects on diabetes mellitus.There were many experimental researches of the mechanism study on the treatment of diabetes mellitus by Huanglianjiedu Decoction.The alkaloid of Huanglianjiedu Decoction acted effectively on diabetes mellitus.It was considered that berberine has significant lipid-lowering and hypoglycemic function.In addition, jatrorrhizine and palmatine were also effective components.In pharmacokinetics,the research of Huanglianjiedu Decoction focused on pharmacokinetic analyse of some kind of medicine components in the normal rats in vivo.In diabetic rats,there were only the pharmacokinetics researchs of baicalin and wogonoside, the research of alkaloids has not been reported. So we suggested that Jatrorrhizine,palmatine,berberine are three effectively alkaloids in Huanglianjiedu Decoction,the pharmacokinetics process of them in normal rats and diabetic rats are still unclear. And we still don't know whether their pharmacokinetics are different in the different body state between physiology and pathology.The second part is experimental study.Earlier publications had described methods on analysis of jatrorrhizine,palmatine and berberine in plasma were HPLC mostly.Compared with LC-MS/MS, HPLC was less sensitive and more time and biomedical samples consuming etc. But a sensitive detection method was a prerequisite to the pharmacokinetic evaluation. So, we plan to establish a sensitive and reliable high-performance liquid chromatography-tandem mass sepctrometry (LC-MS/MS) method on simultaneous determination of jatrorrhizine, palmatine and berberine in rat serum. The optimal ionization and fragmentation conditions,as well as HPLC ones,to detect jatrorrhozine,palmatine and berberine were developed and validated. The detection was performed by MRM mode via electrospray ionization (ESI) source operating in the positive ionization mode.The precursor-to-product ion transitions were at m/z 338→322 for jatrorrhozine , m/z 351.9→308 for palmatine, m/z 336→319.8 for berberine and m/z 356.1→192.1 for tetrahydropalmatine(IS) respectively. The method was linear over the concentration range of 0.2-25 ng·mL^-1 for jatrorrhozine,0.4-50 ng·mL^-1 for palmatine and berberine. The method was validated according to the requirements.It laies the foundation for pharmacokinetic study of jatrorrhizine,palmatine and berberine after oral administration of Huanglianjiedu Decoction.The diabetic model of Sprague-Dawleg rats were established by injecting streptozotocin(STZ) to intraperitoneal. After the Huanglianjiedu Decoction was administered to rats of normal group and the diabetic group through ig route, LC-MS/MS method has been applied to the pharmacokinetic study of the three alkaloids in rat serum. DAS procedure was used to process concentration-time data. To research the pharmacokinetic characters of the three alkaloids in physiology state and pathology state,the results indicate that the pharmacokinetic process of the three alkaloids after oral administration of Huanglianjiedu Decoction were both fitted to be a one-compartment model. But they were different in AUC_0-t, T_max, C_max , MRT_0-t between nomal rats and diabetic rats. The results showed that jatrorrhizine in the diabetic group was absorbed more quickly and got higher serum concentration than rats of normal group; palmatine is more slowly during the process of elimination in the diabetic group; berberine was absorbed better and metabolize more quickly than normal group. It proved that there was different between physiology state and pathology state in pharmacokinetic of the three alkaloids.The experiment was the basis study for pharmacokinetic of Huanglianjiedu Decoction in physiology state and pathology state. Furthermore, it also provided reference for pharmacokinetic study of other compounds of Huanglianjiedu Decoction and material base research of "the same syndrome for Different Disease". In the present study,a highly seletive LC/MS/MS method was developed. It could be modified to make it applicable for determination of other alkaloids of TCM in serum.