The Influence Of Auxiliary Factors On The Chemical Constituents Of Mongolian Medicine And The Principle Of Processing And Attenuation

Caowu has been widely used in clinical,including traditional Chinese medicine,Mongolian Medicine and Tibetan medicine.But unprocessed Caowu contains aconitine type alkaloids with high toxicity,so it is dangerous in clinical use.So raw Caowu only for external use generally,no oral administration unless processed.The traditional experience of Mongolian and Tibetan medicine consider that Chebulae Fructus(Hezi)can detoxify the toxic effects of Aconitum,therefore soaked with Hezi soup is the commonly used method to prepared Zhicaowu in mongolian medicine.Study on this processing method different from traditional Chinese medicine,can provide new ideas for clinical safe and effective use of aconite.ObjectiveFrom different angle to research the influence of processing adjuvants on the quality of Zhicaowu,including 6 kinds of alkaloid content,characteristic fingerprint,toxicity,the rule of dissolving in artificial gastrointestinal fluid and the dynamic change of alkaloid during decocting.Analysis and discussion of relevant results,provide the basis for process optimization and elucidation of reducing toxicity and synergism.Method1.Influence of adjuvant on the content of alkaloids in ZhicaowuProcessing Caowu with different concentration of Hezi soup,the contents of six kinds of alkaloids was determined simultaneously by HPLC.Simultaneously,study the related factors,pH value,the content of tannins and the storage time of Hezi soup.To explore the influence factors of adjuvant on Zhicaowu toxic alkaloids content.2.Study on characteristic fingerprint of ZhicaowuTo develop and validate a high-performance liquid chromatography method for the specific analysis and quantitative determination its index components,analyzed the influence of adjuvant materials processing on the material basis of Zhicaowu.3.Dissolution test in the artificial gastric and intestinal juiceUsing artificial gastric intestinal juice incubation of the powder of raw Caowu and Zhicaowu,and detect the concentration of alkaloids in the samples of different time points,then analysis the influence of Hezi processing on the law of dynamic dissolution of alkaloids.4.Changes of alkaloids in the decoction processDetermination of the content of ester type alkaloids in decoction by HPLC,and analysis of dynamic changes of ester type alkaloids in decoction of Caowu and Zhicaowu.5.The impact on the toxicityFirstly,determination the LD50 of Caowu and Zhicaowu by acute toxicity test on mice to investigate the effect of adjuvant on acute toxicity of Zhicaowu.And then myocardial toxicity and its mechanism of aconitine,chebulagic acidand and their mixture were researched by HCA,analyse the detoxification mechanism of Zhicaowu.Result1.Influence of adjuvant on the content of alkaloids in ZhicaowuWith the increase of the concentration of Hezi soup,the content of total tannins was increasing,pH value decreases at first,then basically stable.The total 6 kinds of alkaloids in processed products increased before they are irregular change.When the Caowu,Hezi ratio was 2:1,the alkaloid content reached the lowest,the ratio was same with Inner Mongolia Mongolian Medicine Standard.2.Study on characteristic fingerprint of ZhicaowuHPLC specific of Caowu prepared with Hezi was established,and 28 common peaks were confirmed,including 17 from Caowu,11 peaks from Hezi.The two new peaks(peak 6 and peak 16)were identified as gallic acid and chebulagic acid by control chromatogram.This HPLC specific analysis method established in this study can be the simultaneously quantitative detection of gallic acid,chebulagic acid,benzoylmesaconine and mesaconitine.3.Dissolution test in the artificial gastric and intestinal juiceNo matter in artificial gastric juice or artificial intestinal juice,mono-ester alkaloids diester-type and alkaloids dissolution rate of Zhicaowu was lower than that of raw Caowu.4.Changes of alkaloids in the decoction processThe diester alkaloids of raw Caowu were hydrolyzed rapidly in the process of water boiling,its content began to decline rapidly in 3 min and disappear after 150 min.Howere,the dister alkaloids of Zhicaowu reached the highest level in 30 min and then decreased slowly.After decocting for 6 hours,the concentration of diester alkaloids was still higher.5.The impact on the toxicityCompared with raw Caowu,the acute toxicity of Zhicaowu reduced by about 30%while content of it dister alkaloids only reduced by 4%.The evaluation results of myocardial toxicity by High content analysis(HCA)is that the concentration of aconitine was 20 mol/L,the mitochondrial membrane potential and intracellular Ca2+ overload were increased,while aconitine and chebulagic acid administered in combination with the phenomenon of calcium overload in myocardial cells and the potential of each dose group of myocardial mitochondrial membrane disappeared,and there was no significant difference between the normal group.ConclusionProcessed by Hezi decoction,the alkaloid content of ZhiCaowu changed compared with raw Caowu,and the chemical composition of the processing materials was introduced.The quantity and quality of components were changed.Hezi has the effect of detoxification on Caowu to a certain extent indeed.howerer,the detoxification principle is different from the traditional Chinese medicine processing method of steaming and boiling,and not to directly reduce the content of diester type Aconitum alkaloids in processing to achieve the role of detoxification.Due to the introduction of the active components of Hezi,such as tannins,therefore solution law of Zhicaowu alkaloids in gastrointestinal digestion is affected to play a role in sustained release(detoxification).In addition,the rate of hydrolysis of diester alkaloids was slowed down in the process of decocting,which played an important role in the retention of the active ingredients.On the other hand,Hezi has strong pharmacological activity,such as cardiac function.The reason that the acute toxicity of Zhicaowu is reduced after Hezi decoction processing is may be the active ingredients such as tannic acid in hezi can relieve the myocardial toxicity of alkaloids in Caowu.