| Backgroud:Chinese medicine plays an important role in traditional chinese medicine.With the modernization of traditional Chinese medicine,Chinese medicine has gradually developed from traditional decoction to formula granule in clinical application.In order to solve the disadvantages of traditional decoction,such as inconvenient carrying and storage,unsuitable for large-scale production,lack of unified standards and so on.But at the same time,the disadvantages of Stewing granules are also obvious,such as confusion in clinical dosage,non-standard production technology,large differences in raw materials and so on.To deal with this problem,the National Pharmacopoeia Committee put forward the concept of "standard Decoction",which is intended to be used as a standard reference to measure whether the Chinese medicine particle prescription（including other forms of decoction pieces）are basically consistent with the clinical decoction.And use modern technical means to determine the characterization parameters of standard decoction and standardize the measurement of modern industrial preparations,so as to achieve the purpose of overall regulation of drug quality.In clinical medicine,Verbena officinalis L is one of the more commonly used traditional Chinese medicinal materials.Its whole grass is used for medicine,which can be taken orally or externally.It is widely used to treat cold,edema,dysentery,jaundice and other diseases.Modern medicine has proved that it has a positive therapeutic effect on inflammation,virus and cancer.However,at present,the relevant quality research standards of Verbena officinalis L are not clear,which is a great obstacle to clinical application.Therefore,the quality research on standard decoction of Verbena officinalis L is of great significance to promote the further development Verbena officinalis L in clinic.Aims:Firstly,the preparation process of Verbena officinalis L standard decoction was controlled and optimized by investigating the parameters such as soaking time,amount of water and decocting time;Secondly,carry out the research on the extraction rate,ethanol-soluble extractives and water content determination of lyophilized powder of Verbena officinalis L standard decoction,take hastatoside and verbenalin as the maker compounds of the Verbena officinalis L standard decoction,and formulate a more appropriate quality research scheme of Verbena officinalis L standard decoction through the study of specific chromatogram and content determination,It provides a reference for the development and utilization of traditional decoction of Verbena officinalis L and other modern application forms of decoction pieces and the formulation of quality standards.to treat cold,edema,dysentery,jaundice and other diseases.Modern medicine has proved thatMethods:1.According to the guiding principles of the item "preparation of standard Decoction" in the technical requirements for quality control and standard formulation of traditional Chinese medicine formula granules（Exposure Draft）,the management specification of traditional Chinese medicine decoction room in medical institutions and the classification of traditional Chinese medicine properties and application parts,the single factor investigation is conducted from three factors:soaking time（0h,0.5h and 1H）,water amount（13,15 and 17 times as much as that of decoction pieces）and decoction time（10min,20min and 30min）,Taking the content of hastatoside,Verbenalin and paste rate as evaluation indexes,the most appropriate preparation process parameters of Verbena officinalis L standard decoction were obtained.Based on these preparation parameters,17 batches of Verbena officinalis L decoction pieces were made into Verbena officinalis L standard decoction.2.Test the paste rate by the following formula出膏率=（浸膏质量（g）×马鞭草标准汤剂总体积（ml））/（马鞭草饮片质量（g）×取样体积（ml））×100%3.The extract of Verbena officinalis L standard decoction was determined according to the hot leaching method in the general rule 2201 extract determination method in the fourth part of Chinese Pharmacopoeia 2020 edition.4.The moisture content of Verbena officinalis L standard decoction lyophilized powder was determined according to the first method of 0832 moisture determination method in the general principle of Part Ⅳ of Chinese Pharmacopoeia 2020 edition.5.Thin layer chromatography was used to detect the standard decoction samples according to the general rule 0502 of the fourth part of the Chinese Pharmacopoeia 2020 edition.6.HPLC was used to optimize the chromatographic conditions in four aspects:Determination wavelength,chromatographic column,test sample solvent and test sample time.After determining the chromatographic conditions,the contents of Verbenalin and hastatoside in Verbena officinalis L standard decoction were determined and the allowable range was calculated.7.The determination wavelength of the specific chromatogram was optimized by ul tra-high performance liquid chromatography.According to the optimized chromatograp hic conditions,specific chromatogram of 17 batches of Verbena officinalis L standard decoction was analyzed.Meanwhile,its specificity,precision,stability and repeatability were investigated.Results:1.The preparation process of Verbena officinalis L standard decoction is as follows:take 100g of Verbena officinalis L decoction pieces,place them in an electric ceramic pot,add water and decoct twice,add 13 times the amount of water in the first decoct,soak for 1h,keep it slightly boiling for 30min after boiling with martial fire,filter the decoction while it is hot by a 350 mesh screen,and quickly cool the filtrate with cold water.Add 11 times the amount of water for the second time,heat it with fire,keep it slightly boiling for 25min after boiling,filter the decoction with 350 mesh screen while it is hot,quickly cool the filtrate with cold water,combine the two decoction,reduce pressure and concentrate at low temperature to 100ml extract;Under magnetic stirring,subpackage into 10ml Brown Xilin bottles.The subpackage volume of each bottle is 2ml.Vacuum freeze-drying,take out and roll aluminum cap.2.The results of TLC showed that the standard decoction of Verbena showed the s ame color spots in the corresponding position as the reference substance and reference c tude drug,which showed good specificity.3.The content range of alcohol soluble extract of 17 batches of Verbena officinalis L standard decoction is 19.77%-51.45%,which meets the requirements;The moisture content of 17 batches of Verbena officinalis L standard decoction is 3.28%-7.5%,which is in line with the regulations.The average extraction rate of 17 batches of Verbena officinalis L standard decoction is 12.8%,and the allowable range is 9.94%15.66%.In the content determination of 17 batches of Verbena standard decoction,the average value calculated by hastatoside is 3.45%,the allowable range is 2.11%-4.79%,the average transfer rate is 78.94%,and the allowable range of transfer rate is 53.28%100%;The average content of verbenolin is 2.25%,the allowable range is 0.07%4.43%,the average transferate is 75.11%,and the allowable range is 49.57%-100%.4.Establish the specific chromatogram of 17 batches of Verbena officinalis L standard decoction,calibrate 5 common peaks,and identify three components such as Verbenalin、hastatoside、verbascoside after comparing with the images of reference substance.Conclusion:The quality on standard decoction of Verbena officinalis L was successfully studied by quantitative analysis of index components and qualitative analysis of specific chromatogram.The methodological investigation proved that the method had good repeatability,high precision and certain standardization.It provides an effective reference for the formulation of the quality standard of Verbena officinalis L standard decoction. |
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