Studies On Quality Characterization Of Jupizhuru Decoction And Gancaoxiexin Decoction

Decoction is a common form of TCMs,but it is not convenient to decoct or carry,and don't have the common standards,which limit its use.So it's necessary to develop standard granule of Chinese classical formulas which is convenient and fit modern life.To achieve this goal,6 companies and many experts participated in the research to develop the granule.The key of the work was how to ensure the granule and the traditional decoction contain the same components,it was also our main research contents.We chose Jupizhuru Decoction as our research object and researched on Jupizhuru Decoction while decocting medicinal herbs in traditional way without any change.Considering the regulations of National Standards and features of TCMs that every decoction contains many chemical constituents and its effect base on all those constituents,after we got the decoction,we decided to develop a quality characterization method combining fingerprint spectrum and determination of index components.Jupizhuru decoction,recorded in Synopsis of Golden Chamber and consisting of tangerine peel,bamboo shavings,jujube,glycyrrhiza,ginseng and ginger,has a wide range of clinical applications and significant effect.Now the company has started to carry out research and development of Jupizhuru decoction,but without record about Jupizhuru decoction or its product in Chinese Pharmacopoeia,here is no method about material characterization or QC of Jupizhuru decoction to learn.According to description of the ancients,blend tangerine peel(15g),bamboo shavings(15g),jujube(12.5g),glycyrrhiza(6g),ginseng(3g)and Ginger(9g)and add 2000 mL of water.After soaking these herbs for 30 minutes,boil the mixture by intense fire to concentrate the water to 600 mL and the total weight is reduced by 1250 grams.The final decoction was processed by precipitation with 75% ethanol and the 75% ethanol and coarse polysaccharide fractions of Jupizhuru decoction were acquired in the end.After the study about the pretreatment method of the medium polarity fraction,the mobile phase,the column temperature and the gradient,we developed the HPLC fingerprints of medium polarity part from the Jupizhuru decoction,which could fully reflect the overall characteristics of the sample.After comparison of the HPLC fingerprints of medium polarity part from the Jupizhuru decoction with reference materials,we identified three characteristic chromatographic peaks.The results include hesperidin,glycyrrhizin and glycyrrhizinate.Then compare that with those of the single herb medicine so that the source of the characteristic chromatographic peaks in the fingerprint of Jupizhuru decoction could be confirmed.And using LC/MS technology,we identify some chemical compositions.As for the coarse polysaccharide fraction of Jupizhuru decoction,its fingerprint was established by the method of overall hydrolysis with TFA,pre-column derivatization with PMP and detection with the HPLC techniques.By comparation to the fingerprint of the standard substance,each chromatographic peakwas identified.The results showed the coarse polysaccharide fraction of Jupizhuru decoction was mainly composed of mannose,rhamnose,galacturonic acid,glucose,xylose,galactose and arabinose,in addition,also contains a small amount of xylose,fucose and glucuronic acid.The method is stable and reliable,could be used to control the quality of the coarse polysaccharide fraction and supplement the HPLC fingerprints of medium polarity part from the Jupizhuru decoction.Referring Chinese Pharmacopoeia,hesperidin,glycyrrhizic acid,ginsenoside Rb1,6-gingerol was chosen as index components.To determinate these index components,we selected the appropriate sample preparation methods to get the test solutions for HPLC analysis.Then we developed determination methods.After study on the methodology,these determination methods proved to be reliable.In order to ensure that the Jupizhuru decoction samples are representative,the single herbs were all from the authentic origin or the main producing areas of herbs.Every kind of single herd was of 2 or 3 origins,and here were not less than 3 batches of the same origin.Randomly combining these herbs,10 batches Jupizhuru decoction samples were got.By determining the similarity between chromatographic fingerprints,yield rate of freeze-dried powder,index components and transfer rate of index components,we can provide reference for the determination of key process parameters in the process of modern technology research.Gancaoxiexin decoction,recorded in Treatise on Febrile Diseases,consists of licorice,Pinellia Ternate,scutellaria,coptis,ginseng,ginger and jujube.According to description of the ancients,blend licorice(12g),Pinellia Ternate(9g),scutellaria(9g),coptis(3g),ginseng(9g),ginger(9g)and jujube(10g)and add 2000 mL of water.After soaking these herbs for 60 minutes,boil the mixture by intense fire to reduce total weight by 750 grams,then throw away all herbs and concentrate the water to 600 mL.then we developed determination methods for glycyrrhizin,baicalin and ginsenoside Rb1.After study on the methodology,we determined index components and transfer rate of these.We developed preparation method of Jupizhuru decoction with the same formulation,dose and decoction technology as these in description of the ancients.Then we developed the HPLC fingerprints of medium polarity part and the coarse polysaccharide fraction both from the Jupizhuru decoction with the help of experience in laboratory.The study on fingerprints clarified material composition primarily.Then we developed determination methods for index components of Jupizhuru decoction and finished study on the methodology.At last,we prepared 10 batches Jupizhuru decoction samples and determined the similarity between chromatographic fingerprints,the yield rate of freeze-dried powder,index components and transfer rate of index components.As for Gancaoxiexin decoction,the determination methods for glycyrrhizin,baicalin and ginsenoside Rb1 were developed and used to determine index components and transfer rate.These studies could help to control quality of modern production of Jupizhuru decoction and Gancaoxiexin decoction.