| ObjectiveThis study was designed to achieve the quality control of Buyang Huanwu Decoction Compound Granules and lay the theoretical foundation for the clinical application and industrial.production,by the content determination of Radix Astragali,Rhizoma Chuanxiong,Radix Paeoniae Rubra and Semen Persic ae.Methods1Extraction conditions of effective parts were determined based on physical and chemical properties,including solvent extraction,solvent concentration,extracting time,extracting pH,etc..Extracts were separated,purified and dried.According to monarch,minister,assistant and guide in TCM prescription and the original proportion of Buyang Huanwu Decoction,extracts were mixed and dried again with appropriate accessories to’ obtain Buyang Huanwu Decoction Compound Granules.2HPLC-ELSD were used to determine the content of astragaloside in Buyang Huanwu Decoction Granule with Agilent EC C18 column(4.6*150mm,4μm),mobile phase of acetonitrile-water(38:62),flow rate of 0.8 mL/min,temperature of ELSD at 80 ℃.The flow rate of atomized gas was 1.6L/min and the gain value was 1.3The contents of tetramethylpyrazine were determined by ultraviolet spectrophotometric method.With the content of tetramethylpyrazine as the evaluation index,the absorbance of tetramethylpyrazine was measured at 254nm.4UV-ELSD were used to determine the content of paeoniflorin with Hypersil C18 column(4.6*250mm,4 μm),mobile phase of acetonitrile-water(16:84),flow rate of 1.0 mL/min.5HPLC-ELSD were used to determine the content of linolein with Agilent Thermo Accucore XL C18 column(4.6*250mm,4μm),mobile phase of acetonitrile-dichloromethane-tetrahydrofuran(52:32:16),flow rate of 1.0 mL/min,temperature of ELSD at 70 ℃.The flow rate of atomized gas was 1.5L/min and the gain value was 1.660 healthy Kunming mice were randomly divided into Bu Yang Huan Wu Decoction Compound Granule(1.574g/ml suspension,prepared according to the maximum concentration)and blank control group by weight and sex,with 30 rats in each group,male and female.Before the formal experiment,the mice were free access to water but not to food.12 hours later,each group of mice were administered 40 mL/kg once a day,with careful observation of activities,eating,drinking and toxic reaction.After the 7 days observation period,the mice were sacrificed and dissected to observe the pathological changes of the main organs,and the weight of the organs were recorded.Result1Astragaloside showed a good linear relationship within the range of 0.20～3.94μg,the average recovery was 102.76%(RSD1.51%).Tetramethylpyrazine showed a good linear relationship within the range of 10～50μg,the average recovery was 100.45%(RSD1.48%).Paeoniflorin showed a good linear relationship within the range of 0.32～4.00μg,the average recovery was 99.97%(RSD1.55%).Linolein showed a good linear relationship within the range of 0.18～3.63μg,the average recovery was 99.92%(RSD1.74%).2In the seven-day observation,the two groups of mice appear no case of death.The appearance,eating,behavior and defecation of mice were nothing abnormal.The growth of body weight is normal as well as the organ index.The differences between Buyang Huanwu Decoction Compound Granule group and the control group was not statistically significant(P>0.05).ConclusionThe content of astragaloside in Buyang Huanwu Decoction Compound Granule is 17.73mg/g.The content of tetramethylpyrazine in Ligusticum chuanxiong hort extract is 0.1045mg/g.The content of paeoniflorin in Radix Paeoniae Rubra extract is 20.25mg/g.The content of linolein in Semen Persicae extract is 1.26mg/g.Buyanghuanwu Decoction Compound Granule dosage for adults is 0.0735g/kg per day,while the maximum dose in this experiment is 63.28g/kg per day,which is 861 times the human clinical dose.No deaths or abnormal acute toxicity occurred.’ Mice had normal increase in body weight during observation period.Anatomy revealed no pathological changes in main organ,indicating that Buyang Huanwu Decoction Compound Granule has no obvious acute toxic effect and clinical application is safe. |
PDF Full Text Request