| Objective: The aim of this project is to establish a perfect quality standard for the herbs of Clerodendrum bungei Steud(CBS),and on this basis,further establish quality standards for the standard decoction and formula granules of CBS,so as to provide a basis for comprehensive and effective control of the quality of herbs of CBS and ensure the safety,reliability and stability of the clinical efficacy of CBS formula granules.Methods:(1)The quality of CBS was evaluated by collecting samples from different sources and based on the requirements of CBS in the 2009 version of the Hunan Province Traditional Chinese Medicine Standards;Through the study on the components of the total phenylethanoid glycosides extraction from CBS,a content determination index components was determined;A HPLC method was established for the determination of verbascoside and the characteristic chromatography for CBS.(2)According to the Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules,18 batches of CBS standard decoctions were prepared.Its traits,identification,content determination and characteristic chromatography were systematically studied,and the quality standard of CBS standard decoctions was established.(3)Take the extraction time,the amount of water added,concentration temperature and the ratio of adjuvants as the investigate factors to conduct a single factor test.And the best process parameters of CBS formula granules were selected based on the extraction rate,the content of verbascoside and the consistency of characteristic chromatography.(4)TLC was used to identify 3 batches of CBS formula granules;The moisture,particle size and solubility of the samples were examined;The content of verbascoside was determined by HPLC,while characteristic chromatography was established to analyze the quality correlation of herbs,standard decoctions and formula granules.Results:(1)All the inspection indexes of 18 batches of CBS herbs meet relevant regulations.A comprehensive qualitative analysis of phenylethanoid glycosides in the total phenylethanoid glycosides extraction from CBS was developed by HPLC-Q-Exactive Orbitrap MS.And the decomposition rules were summarized.A total of 28 phenylethanol glycosides were identified,among which 2 novel phenylethanol glycosides were deduced.The representative component verbascoside was selected as the content determination index.The content determination and characteristic chromatogram method were established by HPLC,and the methodology was investigated.It was stipulated that the content of verbascoside in the herb should not be less than0.2%.6 characteristic peaks were identified in the characteristic chromatogram and 1 peak was identified.(2)18 batches of standard decoctions were prepared,with the extraction rate ranging from 8.0%to 14.4%.The TLC identification method was established,and the unfolding temperature,unfolding humidity and TLC plate were investigated.The standard decoctions showed spots of the same color at the corresponding positions of the chromatograms of the reference herbs.The content of the index components of standard decoctions ranged from 3.70 mg/g to 28.84 mg/g,the transfer rate ranged from 1.6% to 17.8%,and the alcohol-soluble extract ranged from 50.8% to 56.7%.6characteristic peaks were identified in the characteristic chromatogram and 1 peak was identified.(3)Preparation method of CBS formula granules: take 6500 g of CBS decoction pieces,add water to boil,filter,combine the filtrate,concentrate the filtrate to clear cream,add appropriate amount of auxiliary materials,dry(or dry,crush),add appropriate amount of auxiliary materials,mix well,granulate,and make 1000 g.(4)The draft quality standard for CBS formula granules was formulated.TLC was used to identify the CBS formula granules in the corresponding position of the chromatogram with the reference herb,showing spots of the same color.The moisture should not be more than 8.0%.The content determination stipulates that each 1 g of verbascoside should contain 1.8 mg～42.0 mg.The characteristic chromatography indicated 8 chromatographic peaks,it required that the relative retention time of the chromatographic peaks should be in the range of ±10% of specified value.Specified value: 0.43(peak 1),1.09(peak 3),1.09(peak 4),1.09(peak 5)and 1.09(peak 6).1.09(peak 3),1.23(peak 4),1.55(peak 5),1.65(peak 6),1.87(peak 7),2.00(peak 8).The control characteristic chromatography of herbs,standard decoctions and formula granules were constructed,and the 8 common peaks in the formula granules can correspond to the peaks in the herbs and standard decoctions,indicating that the main chemical components of the formula granules can be traced and no new substances were produced during the preparation process.Conclusion: This project initially established a quality inspection system for the herbs,standard decoctions and formula granules of CBS,which can provide a reference basis for the production and quality control of the formula granules. |
PDF Full Text Request