Chemical And Metabolic Study On Aconitum Alkaloids Of Radix Aconiti Lateralls Preparata And Fuzilizhong Pill

Radix Aconiti Lateralls Preparata is a kind of processed product of Aconitum carmichaeli Debx. roots in the secondary root of buttercup, its functions are to cause restoration from collapse, supplement body fire and reinforce yang, dispel wind, cold and damp. The un-processed product of Aconitum carmichaili has strong toxicity, it can be used when it is preprocessed to reduce toxicity. Fuzilizhong pill quoted from the sutra recipe can warm the middle and strengthen the spleen. It consists of five chinese traditional medicines: Radix Glycyrrhizae, Rhizoma Zingiberis, Rhizoma Atractylodis Macrocrphalae, Radix Codonopsis, Radix Aconiti Lateralls Preparata.The project, chemical and metabolic study on aconitum alkaloids of Radix Aconiti Lateralls Preparata and Fuzilizhong Pill, is supported by the ministry of science and technology of the People′s Republic of China.The purpose is to investigate the chemical property and metabolic components of Fuzilizhong Pill and Radix Aconiti Lateralls Preparata. Prepare some alkaloids as reference materials, study their chemical character; detect the metabolites of Fuzilizhong Pill and Radix Aconiti Lateralls Preparata by LC-MS. This thesis is divided into seven parts. These parts are as follows:Firstly, literatures and related researches of the processing history and evolution,the chemical constituents,the pharmacological effect,metabolites and the clinic application of Radix Aconiti Lateralls Preparata and Fuzilizhong Pill are reviewed.Secondly, a high performance liquid chromatography -mass spectrum (MS) analysis method for identification of all aconitum alkaloids and determination of diester-diterpenoid aconitum alkaloids from aconite roots after processing, heating and combination was developed. 20 kinds of aconitum alkaloids were identified .he analysis was carried out with an Agilent C18 at ambient temperature with a sample injection volume of 20μL and the wavelength was at 240 nm. Solvent A(water/ ammonia , 99 : 1 v/v) and solvent B (MeCN) were used as mobile phase (1ml/min). Use a gradient program of 0.00min:A∶B(88∶12V/V;45min:A∶B(38∶62V/V), 60min:A∶B (38∶62V/V).Thirdly, chemical separation was accomplished by using techniques such as supercritical-fluid extraction, column chromatography on silica gel, column chromatography on Sephadex gel, macroporous adsorption resin extraction and LC preparation. 10 compounds were identified with UV, IR, 1H-NMR, 13C-NMR, and by their physical and chemical properties. they were aconitine, mesaconitine, hypaconine, benzoylaconine,β-sitosterol benzoylmesaconine;aconine,hypaconine;N-demethylhypaconitine, N-demethylmesaconitine.Fourthly, rabbits were given orally Radix Aconiti Lateralls Preparata and were killed after administration,and their blood was taken to examine serum metabolites by LC-MS,which revealed that hypaconine ,benzoylaconine,fuziling,nieoline,benzoylmesaconine were absorbed into blood; their urine was taken to examine metabolites by LC-MS, which revealed that hypaconitine,benzoylaconine,fuziling,nieoline were excreted into urine;hypaconitine,benzoylaconine,fuziling,neoline was detected in liver;hypaconitine,benzoylaconine,fuziling,neoline was detected in stomach.Fifthly, optimize the extraction process of Radix Aconiti Lateralls Preparata,using the content of hydrolysate of three kind of diester-type alkaloids(benzoylaconine;benzoylhypaconine;benzoylmesaconine) and diester-type alkaloids(aconitine ,mesaconitine ,hypaconitine)as selection index. The optimum technology of extraction was studied by orthogonal test.The factors of the orthogonal test table L9(34) were ethanol concentration (A),the ratio of material to liquid (B),extracting time(C)and extracting degrees(D).The contents of hydrolysates of three kinds of diester-type alkaloids and diester-type alkaloids were determined by HPLC. This purpose is to find the substitute of Radix Aconiti Lateralls Preparata.Sixth, because the hydrolysates of aconitine, mesaconitine, hypaconitine were complicated. Four hydrolysates of Aconitine were found; Their protonated molecularions at m/z 604,500, 586,482 were monitored; five hydrolysates of mesconine were found; Their protonated molecularions at m/z 590,486,514,618,572 were monitored; four hydrolysates of hypaconine were found; Their protonated molecularions at m/z 604,500,586,482 were monitored. The results improve the cognition of the hydrolysate of these alkaloids.However, the metabolites of aconitine, mesaconitine, hypaconitine in intestinal bacteria metabolite was not complicated. Benzoylmesaconine was prepared by intestinal bacteria metabolite. The method is rapid,easy and effective.Finally, a new method was used to control the quality of Radix Aconiti Lateralls Preparata and Fuzilizhong Pill.