Optimization Of Processing Technology And Hemostatic Mechanism Of Scutellariae Radix Carbonisata

Objective:The best processing technology of scutellariae radix carbonisata was optimized by investigating the three factors of frying temperature,frying time and dosage.The relationship between apparent color and the content of four flavonoids during the processing of scutellariae radix carbonisata was studied.The chemical constituents of Scutellaria baicalensis before and after stir-frying were analyzed by High Performance Liquid Chromatography-Ion Trap-Time-of-Flight Mass Spectrometry（HPLC-IT-TOF）,and the different components were discussed.The mechanism basis of hemostatic action of scutellariae radix carbonisata were studied,so as to provide scientific basis for the processing and clinical application of scutellariae radix carbonisata.Methods:1.Literature research:In this paper,we consulted with the ancient Chinese materia medica and ancient Chinese medical book,analyzed the local traditional Chinese medicine slices processing norms and modern pharmacopoeia.Meanwhile,we reviewed the history of scutellariae radix carbonisata in function and processing,and discussed the modern processing technology,chemical composition and pharmacological research in some extent.We hope it can be helpful for some related research of scutellariae radix carbonisata.2.Optimization of the processing technology of Astragalus charcoal:The experimental design was based on the response surface analysis methods（RSM）,and content of baicalin,the clotting time of the rats and the external properties of scutellariae radix carbonisata were used as the response values.3.A Study on the relationship between the color characteristics of the processing of Scutellariae Radix Carbonisata and the contents of four flavonoids:The chromaticity space parameters of the samples during the processing of Scutellariae Radix Carbonisata were measured by the visual analyzer to analyze the color variation rule during the processing of Scutellariae Radix Carbonisata,and the content changes of baicalin,wogonoside,baicalein,wogonin,Pearson correlation analysis and linear regression analysis of the data were carried out by SPSS 20.0 software.4.Blood coagulation mechanism:rats were given intragastric administration of scutellariae radix carbonisatac and Scutellaria water decoction respectively.The changes of anal temperature,Thrombin Time（TT）,Prothrombin Time（PT）,Activated Partial Thrombin Time（APTT）,Plasma Fibrinogen Time（FIB）,whole blood viscosity,red blood cell count（RBC）,hemoglobin（HGB）content,platelet count（PLT）,white blood cell count（WBC）and lung pathology were measured.5.Study on the chemical composition of scutellariae radix carbonisata before and after stir-frying:chromatographic column:Yilit C₁₈（250 mm×4.6 mm,5μm）;mobile phase:acetonitrile（A）-pure water（B）,gradient elution,flow rate 1 m L/min;injection volume 5μL).The electrospray ion source was selected to collect and detect in positive and negative ion mode,and the data was analyzed by LC-MS solution software.The components were identified according to the retention time,accurate relative molecular weight and fragmentation ion information provided by MS,and combined with the reference substance and literature information.After normalization,the mass spectrometry data of each sample were analyzed by principal component analysis（（PCA））and orthogonal partial least square discriminant analysis（OPLS-DA）before and after stir-fried charcoal from scutellariae radix.Results:Through comprehensive evaluation,Optimal processing parameters were as follows:frying 10 min;the bottom temperature at 200℃;the dosage is 100g.The experimental results of the changes of chromaticity spatial parameters and the contents of four flavonoids showed that during the processing of Scutellariae radix carbonisata,the contents of baicalin and wogonin showed a downward trend,while baicalein and wogonin increased at first and then decreased.The results of Pearson correlation analysis showed that the chromaticity spatial parameters L*（luminance value）,a*（red-green value）,b*（yellow-blue value）and E*ab（total color value）were significantly correlated with the contents of baicalin,wogonin,baicalein and wogonin（P<0）.01),in which the total color value E*ab was significantly positively correlated with the contents of baicalin and wogonin,and negatively correlated with the contents of baicalein and wogonin.The results of linear regression analysis also showed that there was a strong correlation between them.The experimental results of blood coagulation mechanism showed that compared with the blank group,the anal temperature,the whole blood high,middle and low shear viscosity and TT,APTT,PT of the model group increased significantly,the whole blood PLT,RBC and HGB increased significantly,and WBC decreased significantly（P<0.05）.The whole blood high,medium and low shear viscosity of the model group increased significantly（P<0.05）.P<0.01).Scutellariae radix carbonisata and its fried charcoal products could significantly reduce the anal temperature of rats.Scutellaria baicalensis group could significantly shorten the PT and APTT time,TT time and FIB content of blood heat hemorrhage model rats.Scutellaria charcoa group could significantly shorten the PT,APTT(P,FIB content and TT time of blood-heat hemorrhage model rats.Scutellaria baicalensis group and Huangqin carbon group could significantly reduce the whole blood high and middle shear viscosity of blood-heat hemorrhage model rats（P<0.05,P<0.01）.Scutellaria baicalensis group could significantly reduce the low shear viscosity of blood-heat hemorrhage model rats,but there was no significant difference between Scutellariae radix carbonisata group and Scutellaria charcoal group.Scutellaria baicalensis group and Huangqin carbon group could significantly increase the contents of PLT and WBC,and decrease the contents of RBC and HGB in blood-heat hemorrhage model rats（P<0.05,P<0.01）.A total of 57components were identified from Scutellaria baicalensis extract and 18 components were identified from Scutellariae radix carbonisata extract.The results of PCA and OPLS-DA showed that there were significant differences in the content of chemical constituents of Scutellaria baicalensis.Scutellariae Radix Carbonisata can reduce pulmonary bleeding,inflammatory infiltration of inflammatory cells,alveolar cell wall thickening has obvious therapeutic effect.Conclusion:Through the study of this subject,the best processing technology of Scutellariae radix carbonisata was confirmed,according to which the appearance and internal quality of Scutellaria charcoal were uniform and stable.Through the pharmacodynamic experiment,it is proved that scutellariae radix carbonisata has the effect of cooling blood and stopping bleeding.Through HPLC-IT-TOF-MS analysis,the chemical composition of Scutellaria baicalensis before and after charcoal frying is indeed significantly different,and the loss of components and mutual transformation between components caused by high temperature are the main reasons for the differences.