| OBJECTIVE: Polygonum divaricatum L.,a Mongolian medicinal herb,is a dry root or whole grass of Polygonum divaricatum L.It is sour,bitter and astringent,as well as cool,thin,light,sputum and blunt.It has the effects of clearing away heat and stopping diarrhea.It can also treat enteritis,dysentery,vomiting,osteoporosis,water and fire scald and various tumors after compatibility.And the clinical effect is very remarkable.In addition to being applied to the group,it is also expected to be developed into a drink as a common wild vegetable.However,as a medicinal material,there is rarely a report on the safety of its consumption.Therefore,in this study,Polygonum divaricatum L.was used as the research object,and the flavonoids were extracted from this herb by reflux extraction technique.The extraction and purification process was optimized,and the isolated flavonoids were identified.It also conducts oral and acute toxicity and subchronic toxicity research in vivo,which provides a test basis for the further development of Polygonum divaricatum L.,and contributes to solving the shortage of Mongolian medicine resources and sustainable development.METHODS: TFP were extracted by reflux extraction technique.And the total flavonoids extraction process was optimized by using TFP content as the index.The purity of TFP was taken as the index and the total surface area was optimized by column chromatography combined with response surface methodology to purified the flavonoids.Then the TFP were separated to obtain monomer compound,and the structure of the monomer compound is determined by UPLC-MS/MS technology.The in vivo safety of TFP was evaluated by acute toxicity test and repeated dose 28 days oral toxicity test in rats.RESULTS: The optimal extraction process were: ethanol concentration 70%,ratio of material to liquid 1:10,extraction time 2.2 h,extraction temperature 74 °C,TPF content was 2.42±0.05 %;optimal purification process were: 22 mL of the extract,adjust the pH to 5,and elute with 65% ethanol at a flow rate of 2 mL/min to obtain a purity of 39.88 % ± 0.19 % of TFP.After that,the TFP was separated and the parent ion and the second fragment ion of the compound were obtained by UPLC-MS/MS technology.The structure of the monomer compound was determined by comprehensive software database and literature reports.As a result TFP mainly contained 7 flavonoids:Quercetin,Myricetin,Kaempferol,Avicularin,Hyperoside,Isoquercitrin and Rutin.The acute toxicity and subchronic toxicity of the test were observed by feeding different doses of the aqueous extract of Polygonum divaricatum L..The results showed that there was no obvious abnormality in behavior,activity,diet and excretion of mice in each dose group in the acute toxicity test.There was no significant difference between the blank group and the control group(p>0.05).There were no abnormalities in the color,size and proportion of the important organs(heart,liver,spleen,lung and kidney)of the mice.And no death of the mice occurred.According to the value of the acute toxicity dose classification table,it was determined that Polygonum divaricatum L.were not poisonous.In the repeated dose 28 days oral toxicity test,no abnormalities were found in the mental activity,eating and drinking,and excretion of the rats.There was no significant difference in index change(p > 0.05).No death occurred in 28 days,hematological parameters(hemoglobin,red blood cells,white blood cells,neutrophils,monocytes,lymphocytes)and blood biochemical indicators were not significantly different from the control group(p> 0.05).CONCLUSION: It is a high-quality Mongolian medicine resource that can be utilized.Polygonum divaricatum L.,the Mongolian herba,mainly contained flavonoids.Acute toxicity and short-term toxicity were not observed in Polygonum divaricatum L.,indicating that they were basically safe at normal doses. |
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