| Background: China has paid great attention to the industry of traditional Chinese medicine in China.In recent years,China has issued a number of policies and regulations related to the famous classical formula,,providing a good policy environment for the research of the famous classical formula.Daqinjiao decoction was included in the Catalogue of Ancient famous classical formulas(the First Batch),which is widely used in clinical practice.The study on its substance benchmark quality can lay an important foundation for the study of its compound preparations in the later stage.Objective: In order to ensure the consistency between the famous classical formula and the ancient medical books and other references,the literature about the famous classical formula "Daqinjiao decoction",such as the medicinal material source,dose and the decocting process was determined in the reference study.To lay a foundation for the establishment of the substance benchmark of Daqinjiao decoction and its further Research and Development study,it was needed to study the major preparation methods of Daqinjiao decoction and quality control method of the substance benchmark of Daqinjiao decoction,as well as the process of quantity value transfer.Methods: 1.Study on preparation of substance benchmark of Daqinjiao decoction:(1)Prescription textual research: Based on the historical evolution of prescription in ancient literature,and combined with modern literature records,textual research was conducted on the basic source,prescription dosage and decocting technology of various individual medicinal materials in Daqinjiao decoction.(2)Preparation technology research: based on the prescription research results,using the single factor investigation,to determine the decoction method according to the decoction extract rate,the transfer rate for gentiopicrin and loganic acid,the processing of the decoction can be decided.2.Quality study:(1)To establish medium polarity fingerprint and content determination method of Daqinjiao decoction by UPLC method.(2)To establish the low polarity fingerprint and content determination method of Daqinjiao decoction by UPLC method.3.Determination of substance reference in 15 batches of Daqinjiao decoctionLots batches of the herbal slices was collected,which were line with the2015 edition of "Chinese pharmacopoeia" in the quality control.By random matching with these batchs of the herbal slices,15 batches of the substance benchmark were prepared.Vacuum freeze drying was used to get 15 batches of the corresponding products of dry substance benchmark.4.Quantity-value transfer researchTo study the quantity transfer process of decoction pieces to material benchmark,the indexes were used such as the extraction rate and the transfer rate of 10 index components,such as loganic acid,gentiopicrin,prim-O-glucosylcimifugin,paeoniflorin,glycyrrhiza,baicalin,glycyrrhizic acid,osthol,asarinin and columbianadin.Results:1.Study on preparation of substance benchmark of Daqinjiao decoction:Daqinjiao decoction is a powder in the collection of the mechanism of disease,Su Wen Bing Ji Qi Yi Bao Ming Ji compiled by Liu Wansu in the Jin Dynasty.The medicinal herbal materials used are all within the source range prescribed by the 2015 edition of Chinese Pharmacopoeia.Preparation process: weighed(roughly meshing for through 5 ～ 8 order sereen)Gentianae Macrophyllae Radix 5.28 g,Glycyrrhizae Radix et Rhizoma 3.52 g,Chuanxiong Rhizoma 3.52 g,Angelicae Sinensis Radix 3.52 g,Paeoniae Radix Alba 3.52 g,Gypsum Fibrosum 3.52 g,Angelicae Pubescentis Radix3.52 g,Notopterygii Rhizoma et Radix 1.76 g,Saposhnikoviae Radix 1.76 g,Scutellariae Radix 1.76 g,Angelicae Dahuricae Radix 1.76 g,Atractylodis Macrocephalae Rhizoma 1.76 g,Rehmanniae Radix Recens 1.76 g,Rehmanniae Radix Preparata 1.76 g,Poria 1.76 g,Asari Radix et Rhizoma0.88 g.Add 600 m L water to the above 16 herbs slices and place them in a 2L ceramic pot.Bring to a boil on high heat(500 watts)and continue to simmer(300 watts)for about 50 minutes until the decoction reaches 300 m L.After sieving with 400 mesh gauze,the decoction was freeze-dried in vacuum to obtain the substance benchmark corresponding to the material freeze-dried powder of Daqinjiao decoction.2.Quality study:(1)The medium polarity UPLC fingerprints of Daqinjiao decoction and the method for determining the content of 8 components were established.39 common peaks were identified,and 11 components were identified,including loganic acid,gentiopicrin,albiflorin,paeoniflorin,ferulic acid,prim-O-glucosylcimifugin,glycyrrhiza,5-O-methylvisalaigoside,baicalin,wogonoside,and glycyrrhizic acid.The contents of 8 index components,such as loganic acid,gentiopicrin,prim-O-glucosylcimifugin,paeoniflorin,ferulic acid,glycyrrhizin,baicalin and glycyrrhizic acid,were determined.(2)Low polarity UPLC fingerprints of Daqinjiao decoction and the method for determining the content of 3 components were established and signals at 245 nm and 350 nm were collected.Twenty common peaks were identified at 245 nm,and four components were identified,including osthol,notopterol,asarinin and columbianadin.The content of asarinin was determined.Ten common peaks were identified at 350 nm.The contents of osthol and columbianadin were determined.3.Determination of substance reference in 15 batches of Daqinjiao decoctionThe extract rate of 15 batches of decoction ranged from 30.62% to33.62%,the moisture of 15 batches of freeze-dried powder ranged from4.29% to 7.35%,and the extract ranged from 46.63% to 50.85%.The similarity of the fingerprints of the 15 batches of material reference medium polarity UPLC was > 0.969.The similarity of low polarity UPLC fingerprints at 245 nm was > 0.841,and the similarity of 15 batches of samples at 350 nm was > 0.962.The mass fraction ranges of 11 components in 15 batches of substance benchmark were as follows: loganic acid 2.63 ～5.99 mg/g,gentiopicrin 8.13 ～ 19.63 mg/g,paeoniflorin 3.82 ～ 5.18 mg/g,ferulic acid 0.35 ～ 0.50 mg/g,prim-O-glucosylcimifugin 0.16 ～ 0.31 mg/g,baicalin 0.39 ～ 1.75 mg/g,7.45 ～ 12.09 mg/g,glycyrrhizic acid 0.70 ～ 2.16mg/g.osthol 50.82 ～ 76.11 μg/g,columbianadin 10.99 ～ 15.08 μg/g,asarinin7.07 ～ 11.17 μg/g.4.Quantity and quality transfer research:(1)In the 15 batches of substance benchmark,the transfer rates of 10 components from decoction pieces to material reference were as follows:loganic acid 46.04% ～ 87.62%,gentiopicrin 51.80% ～ 83.71%,paeoniflorin47.59% ～ 60.05%,prim-O-glucosylcimifugin 53.65% ～ 82.97%,glycyrrhiza19.03% ～ 63.31%,baicalin 40.23% ～ 60.29%,lycyrrhizic acid 10.35% ～32.83%,osthol 30.38% ～ 44.18%,asarinin 45.82% ～ 65.66%,columbianadin13.49% ～ 19.44%.(2)The paste yield transfer rate of 15 batches of materials was 79.36%～ 92.86%.Conclusion: Based on literature research and actual decocting data,the preparation technology of material reference of Daqinjiao decoction was stable.The two sets of medium and low polarity fingerprints and the content determination method of 11 components established by UPLC are accurate and feasible,and the quantity transfer process of decoction from herbal material slices to its substance benchmark of the Daqinjiao decoction was determined.The above has laid good foundation for establishment and further research and developmend of the famous classical formula Daqinjiao decoction. |
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