Comparison Of Chemical Constitution,Acute Toxity And Anti-inflammatory Activity Between Single Decoction Of Radix Sophorae Tonkinensis And Its Compound Decoction

Radix Sophorae Tonkinensis is dried root and rhizome of Sophora tonkinensis Gapnep,which has effect of heat-clearing,detoxifying,diminishing swelling and relieving sore throat.It can be used for treating diseases such as swelling and pain in throat and gingiva,acute tonsillitis,bronchitis,pneumonia,chronic active hepatitis,tumor and arrhythmia,etc.Radix Sophorae Tonkinensis belongs to toxic Chinese medicine.It mainly contains chemical components of alkaloids,flavonoids,and triterpenoid saponins,in which,alkaloids are not only its effective constituents,but also its toxic ingredients.There are many reports about the toxic,side effects and adverse reactions of Radix Sophorae Tonkinensis,which greatly limits its clinical application.Drug compatibility is a notable characteristic of traditional Chinese medicine,whether the toxicity reducing and efficacy enhancing(efficacy maintaining)can be achieved by compatibility for Radix Sophorae Tonkinensis,is deserved to be focused and studied.In this paper,the single decoction of Radix Sophorae Tonkinensis(SD-RST)and the mixed decoction of Radix Sophorae Tonkinensis with Rhizoma Belamcandae and Rhizoma Cimicifugae(MD-RST/RB/RC)were taken as the research objects.The qualitative and quantitative comparison of chemical constitution were studied from the aspects of chemical composition,the content of main chemical components,and the content of total alkaloids and total phenols between SD-RST and MD-RST/RB/RC.Meanwhile,the acute toxicity and anti-inflammatory activity of SD-RST and MD-RST/RB/RC were also comparatively studied.It would make an attempt to reveal the correlation of chemical components,toxicity,and pharmacological activity,and to provide scientific basis for safe and effective clinical medication of Radix Sophorae Tonkinensis.The paper is divided into several parts as follows.(I)The SD-RST and MD-RST/RB/RC were prepared by using decoction method commonly used in clinic.Then,their chemical compositions were qualitatively compared by UPLC-Q TOF MS method.35 alkaloids compounds were found from SD-RST and MD-RST/RB/RC including 23 alkaloids in SD-RST and 28 alkaloids in MD-RST/RB/RC.Wherein,7 alkaloids were unique in SD-RST and 12 alkaloids were unique in MD-RST/RB/RC.16 compounds were the same components in SD-RST and MD-RST/RB/RC.The PCA analysis showed that the two groups of SD-RST and MD-RST/RB/RC samples were intra-group gathered and inter-group separation in sample distribution scatter diagram,which demonstrated that there were significant differences of the alkaloids components between SD-RST and MD-RST/RB/RC.In addition,the difference of 10 alkaloids compounds such as oxymatrine,matrine,and sophoranol have been found from the component distribution scatter diagram.The alkaloids part and non-alkaloid part of SD-RST were prepared by using macroporous resin and polyamide resin and identified by the UPLC-Q TOF method.It has been found that the kinds of alkaloids constitunts contained in the alkaloid part and SD-RST were basically the same.35 compounds were qualitatively obtained from the non-alkaloid part of the SD-RST,including 19 flavonoids and 14 saponins compounds.The non-alkaloid part of the SD-RST is mainly composed of flavonoids and saponins.(?)An HPLC method was developed for simultaneous determination of seven alkaloids(cytosine,oxymatrine,N-oxysophocarpine,N-methylcytisine,sophoranol,matrine,and sophocarpine)and three flavonoids(trifolirhizin,fermononetin,and maackiain).For the method validation,these ten compounds showed good separation and satisfactory linearity(r?0.999 7)within the concentration ranges tested.The mean recoveries were in the range of 98.6%to 102.6%with the RSD(n = 6)between 0.68%and 3.66%.This method was proved to be simple,accurate and repeatable.The contents of 10 main components have been determined in the Radix Sophorae Tonkinensis materials,SD-RST and MD-RST/RB/RC by this method.The quantitative results showed that there were significant differences in the contents of seven alkaloids and three flavonoids among the different Radix Sophorae Tonkinensis materials.In the decoction process of 3 batches of Radix Sophorae Tonkinensis materials,the total transfer rate of 10 ingredients were 53%,64%and 55%,respectively.The content of oxymatrine,N-oxysophocarpine,cytosine,N-methylcytisine,formononetin,maackiain and trifolirhizin were decreased,but the content of matrine,sophoranol,and sophocarpine were increased.It demonstrated that the content differences of 10 components between Radix Sophorae Tonkinensis materials and SD-RST,were related to not only the component transfer rate,but also the chemical reactions in the decoction process.Furthermore,the content of the 6 alkaloids in MD-RST/RB/RC were decreased,compared with that of SD-RST,wherein,oxymatrine had the largest decreasing amplitude and matrine had the smallest decreasing amplitude.It demonstrated that the mixed decocting of Radix Sophorae Tonkinensis with Rhizoma Belamcandae and Rhizoma Cimicifugae,could promote the change of alkaloids constituents.There were some differences of the main alkaloids between SD-RST and MD-RST/RB/RC.(?)The contents of total alkaloids and total phenols were determined by acid dye colorimetry and ferric trichloride-potassium ferricyanide system,respectively.The results showed that the extraction rate of total alkaloids was higher than that of total phenols in the process of decocting of Radix Sophorae Tonkinensis.The total alkaloids of MD-RST/RB/RC were less than that of SD-RST.It could further demonstrate that the alkaloids can be decomposed and transformed to other compounds in the decocting process.The total phenols of MD-RST/RB/RC were higher than that of SD-RST.It may be related to the presence of phenolic compounds in Rhizoma Belamcandae and Rhizoma Cimicifugae.(IV)In order to compare the toxicity effect of SD-RST with MD-RST/RB/RC and negative control decoction(Rhizoma Belamcandae and Rhizoma Cimicifugae were decocted mixedly.),the acute toxicity of mice was studied.It showed that the death rates of mice in SD-RST groups for each dosage were all higher than that of MD-RST/RB/RC groups and the negative control groups had no death records.Calculated in Radix Sophorae Tonkinensis materials,the LD50 of SD-RST and MD-RST/RB/RC were 19.6 g/kg and 31.7 g/kg,respectively.It demonstrated that MD-RST/RB/RC had toxicity attenuation effect.(V)The model of mice's auricle swell caused by dimethylbenzene was used to compare the anti-inflammatory activity of SD-RST with MD-RST/RB/RC and negative control decoction.The results showed that they all had obvious anti-inflammatory activity compared with the model control groups.The anti-inflammatory activity of the negative control decoction was no significant difference with that of SD-RST and MD-RST/RB/RC(P>0.05).There was no significant difference(P>0.05)between SD-RST and MD-RST/RB/RC for their anti-inflammatory activities.Therefore,MD-RST/RB/RC had protective effect of anti-inflammatory activity.In summary,the comparison of chemical constitution,toxicity and anti-inflammatory activity between SD-RST and MD-RST/RB/RC were studied.It could provide scientific evidences to reveal the correlation among chemical compositon,toxicity,and pharmacological activity,and to explain the compatibility for reducing toxicity and enhancing or maintaining efficacy.