Study On Quality Standard And Pharmacodynamics Of Qingfei Yucuo Granules

Objective:Qingfei Yucuo Recipe is an experiential recipe of my tutor,which is used to cure acne.It found on the base of his clinical experience and climatic environment of Lingnan region.The recipe includs Scutellaria root 15g,Red peony root 10g,,Loquat leaf 15g,ect.Its efficacy is heat-clearing,damp-drying drug,and dispersing stasis to dissipate blood stasis.In the early stage of this task,the dosage forms of Qingfei Yucuo Recipe was improved and the preparation technology of granules was studied,in order to make the recipe easier to be take.To provide basis for quality control of Qingfei Yucuo Formula and its granules,this study intends to establish a method for the determination of baicalin and tanshinone IIA in Qingfei Yucuo Granules and complete its methodological verification.At the same time,qualitative identification of Radix Scutellariae,Radix Salviae Miltiorrhizae and Radix Paeoniae Rubra in Qingfei Yucuo Granules was carried out to investigate the influencing factors.To clarify the mechanism of therapeutic effect that Qingfei Yucuo Granules cure acne,it is going to explore both the anti-inflammatory effect and Immunomodulatory effect of Qingfei Yucuo Granules in mice.Methods:In this study,TLC was used to identify Scutellaria baicalensis,Salvia miltiorrhiza and Paeonia rubra in Qingfei Yucuo Granules,and the factors affecting the results of TLC were investigated.The content of baicalin and tanshinone IIA were determined by high performance liquid chromatography,and the method was investigated from the aspects of specificity,precision,repeatability and stability.Preparation of Qingfei Yucuo Granule intermediate based on the optimum extraction and purification technology.Inflammatory mice model,rat model of cotton ball granuloma and cytoxan-induced immunosuppressed mice were used to explore the anti-inflammatory effect and imunomodulatory effect of Qingfei Yucuo Recipe.Result:The TLC developing solvent for qualitative identification of Scutellaria baicalensis is ethyl acetate-butanone-formic acid-water(10:3:1:1);sample size is 15μL,;saturated at room temperature;saturation time is 30 min,;color developer is 1%FeCl3-ethanol solution.The TLC developing solvent for qualitative identification of Salvia miltiorrhiza is petroleum ether-ethyl acetate(5:1);sample size is 15μL;saturated at room temperature;saturation time is 30 min.The TLC developing solvent for qualitative identification of Radix Paeoniae Rubra is ethyl acetate-formic acid-glacial acetic acid-water(15:1:1:2);sample size is 15 μL,;saturated at room temperature;saturation time is 60 min;spray 5%vanillin sulfuric acid solution and heat it in an oven at 105 C for 3 minutes to develop color.The chromatographic conditions used for quantitative determination of baicalin were mobile phase:methanol-0.376%phosphoric acid solution(47:53);flow rate:1.0 mL.min-1;detection wavelength:280 nm;injection volume:10 μL.Quantitative determination of tanshinone Ⅱ A content is based on mobile phase:acetonitrile-0.02%phosphoric acid solution,gradient elution according to the method of determining tanshinone II A in Chinese Pharmacopoeia 2015 edition;flow rate is 1 mL.min-1;column temperature is 20℃;detection wavelength is 270 nm,injection volume is 100μL.The results of pharmacodynamics test showed that there was no significant change in OD value of peritoneal cleaning fluid in mice in each dose group of Qingfei Yucuo Granule.The wet and dry weight of cotton ball granuloma in each dose group of Qingfei Yucuo Granule did not change significantly compared with the model group.The moderate and high doses of Qingfei Yucuo Granule could obviously increase the phagocytic index of model animals,and the OD value of serum hemolysin in mice in middle and high doses of Qingfei Yucuo Granule decreased significantly.Conclusion:In TLC identification,the spots are clear and orderly,the separation is good,and the method is simple.It can be used for qualitative identification of Scutellaria baicalensis,Salvia miltiorrhiza and Paeonia lactiflora in Qingfei Yucuo Granules.The established method is simple and rapid.The peak shapes of baicalin and tanshinone IIA in the samples are good,and the methodological evaluation meets the requirements except the recovery rate of tanshinone IIA.Pharmacodynamic results showed that Qingfei Yucuo Granule had weak Anti-Acute exudative inflammation and chronic proliferative inflammation.The middle and high dose groups had obvious improvement on the non-specific immunity reduction induced by cyclophosphamide in mice,but had inhibitory effect on specific humoral immunity in mice.The specific mechanism needed further study.