The Substance Benchmarks Research And Development Of Classical Prescription Danggui Buxue Decoction

Danggui Buxue Decoction,recorded in Nei Wai Shang Bian Huo Lun,is an ancient classic prescription for replenishing qi and blood,and composed of Astragali Radix and Angelicae Sinensis Radix in ratio of 5:1.There are a multitude of modern researches on Danggui Buxue Decoction,but due to the different quality and preparation procedures,the quality of Danggui Buxue Decoction varies greatly,and the Decoction quality could not be compared horizontally.Therefore,it is necessary to develop a unified standardized preparation method of the Decoction,and establish the quality standard of substance benchmarks.To solve the problem,the following researches have been performed in this thesis:1.Textual research on the history of the prescription was carried out to make clear the prescription information such as the dosage of the prescription,the origin of the medicinal materials and the processing method.2.Research on medicinal materials and the preparation process of substance benchmarks were conducted.The resources of Astragali Radix and Angelicae Sinensis Radix were the primary factors to be considered in optimizing the quality of the Decoction.According to the herbalogical study,both herbs are originated from the roots of Astragalus membranaceus(fisch.)Bge var.mongholicus(Bge.)Hsiao and Angelica sinensis(oliv.)Diels respectively.In order to ensure the representative selection of Chinese herbal medicines,at least 15 batches of Chinese herbs from three places of origin were collected for quality analysis.Finally,the Angelicae Sinensis Radix from Minxian county and Astragali Radix from Weiyuan county were found to be the best in quality.In addition,the preparation procedure of substance benchmarks were investigated.The effects of decocting,concentrating and drying methods on the herbal drug standardwere investigated by comparing extraction rate and the transfer rate of ferulic acid,calycosin7-O-β-D-glucopyranoside,astragaloside Ⅳ and polysaccharide.The optimal preparation conditions were determined as follows:soak for 30 minutes,add 10/8 times of water,decoct twice,1/0.5h,concentrate at 50℃and freeze-drying.3.The quality control method of substance benchmarks were established.A thin layer chromatography identification method was established,with which the exclusive identification of Astragali Radix and Angelicae Sinensis Radix could be achieved in the substance benchmarks of the Decoction.A HPLC method for simultaneous determination of ferulic acid and calycosin7-O-β-D-glucopyranoside in the substance benchmarks was established,and a HPLCELSD method is established to realize the quality control of astragaloside Ⅳ.A HPLC-ELSD fingerprint method of the Decoction was used to evaluate the quality consistency of the Decoction,and the chemical profile of the Decoction was characterized comprehensively.All the above methods were validated by rigorous methodology including precision,accuracy,linearity and range,stability,specificity and durability.The established methods can be used for the quality control of the Decoction.4.The chemical constituents of the Decoction were analyzed by reversedphase chromatography and hydrophilic interaction chromatography coupled with quadrupole timeof-flight mass spectrometry,through the in-house library,HMDB database,Reaxys database and comparison with reference substances.In summary,a total of 154 compounds were identified and characterized,including 16 saponins,44 flavonoids,10 phthalides,7 phenylpropanoids,15 nucleosides,30 sugars and 32 other compounds.