| Objective:Under the guidance of traditional Chinese medicine theory,this thesis,based on the famous classic prescription,Houpowenzhong decoction,and its material benchmarks,using modern analytical technologies and methods synthetically as well as combining relevant national policies,develops the study on quality standard about raw herbal materials and decoction pieces,which helps to clarify the source and origin of this medicinal material.Furthermore,comprehensive quality control method for Houpowenzhong decoction was established,which contributes to control the quality of products effectively on the whole as well as provides a preferable technical foundation to ensure the stability and safety of clinical effect.Methods:1.Study on quality standards of raw materials(1)Collection of raw medicinal materials.Eight different herbs including magnolia officinalis,pericarpium citri reticulatae,alpinia katsumadai,costus root,rhizoma zingiberis,licorice,poria and ginger were collected from different regions and batches.(2)Analysis of raw materials.According to the requirements of the Chinese Pharmacopoeia in 2015,the collected eight raw materials were identified and examined.(3)Study on fingerprint pattern of raw material.Using Ultra Performance Liquid Chromatography(UPLC),study on the fingerprint of raw materials in different production areas and various batches were carried out.After optimization,the preperation method and chromatographic conditions of the samples were identified with methodology.By means of fingerprint similarity software by the State Pharmacopoeia Committee,the date including common feature peaks,relative retention time and the relative peak area would apply to establish the UPLC fingerprint of raw materials.2.Study on quality standards of decoction pieces(1)Inspection decoction pieces.The tests were performed in accordance with the requirements of the Chinese Pharmacopoeia in 2015 for each piece of decoction,and the samples that passed the test were included in subsequent studies.(2)Study on fingerprint of decoction pieces.Using Ultra Performance Liquid Chromatography(UPLC),study on the fingerprint of decoction pieces in different production areas and various batches were carried out.After optimization,the preperation method and chromatographic conditions of the samples were identified with methodology.3.Material benchmark research of Houpowenzhong decoction(1)Qualitative identification of thin layers.The thin layer chromatography(TLC)method was used to qualitatively identify the corresponding objects of Houpowenzhong decoctio and conduct methodological investigations.(2)Study on assay.Using magnolol,honokiol contained in monarch drug,magnolia officinalis and hesperidin contained in minister drug,tangerine peel as indicators,HPLC method was used to establish the content determination methods,and methodological investigations were carried out.Study of fingerprints.The ultra-high performance liquid chromatography(UPLC)method was used to establish the UPLC fingerprints of Houpowenzhong decoction,and the methodology was investigated.Analysis was performed according to the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System"(2004 edition),and the common chromatographic peaks were identified for components and medicinal materials.Results:1.Study on quality control of raw materials(1)A total of 15 batches of magnolia officinalis,tangerine peel,alpinia katsumadai,radix auckladiae,dried ginger,licorice,poria and ginger were collected,and all of them met the relevant regulations under the 2015 edition of the Chinese Pharmacopoeia.(2)The UPLC fingerprints of the ingredients of Houpowenzhong decoction,including magnolia officinalis,tangerine peel,alpinia katsumadai,radix auckladiae,dried ginger,licorice,poria and ginger were established.Each common chromatographic peak in the established UPLC fingerprints have a high separation nate and strong specificity.2.Study on quality control of decoction pieces(1)Collect 15 pieces of magnolia officinalis,tangerine peel,alpinia katsumadai,radix auckladiae,dried ginger,licorice and poria pieces,and all of them meet the relevant regulations under the 2015 edition of the Chinese Pharmacopoeia.(2)The UPLC fingerprints of Houpowenzhong decoction pieces,including magnolia officinalis,tangerine peel,alpinia katsumadai,radix auckladiae,dried ginger,licorice,poria and ginger were established.Each common chromatographic peak in the established UPLC fingerprints have a high separation nate and strong specificity.3.Material benchmark research of Houpowenzhong decoction.(1)Established thin-layer identification methods for ginger magnolia officinalis,tangerine peel alpinia katsumadai,fried licorice,radix auckladiae and dried ginger,and identified characteristic spots of magnolol,honokiol,hesperidin,pinocembrin,glycyrrhetinic acid,costunolide,dehydrocostuslactone and 6-gingerol.(2)The UPLC fingerprints of the corresponding objects of Houpowenzhong decoction were established and and 12 common characteristic peaks were identified.Compared with the Reference Chromatogram,8 chromatographic peaks were identified,which are 3#rutin naringin and 4#hesperidin,5#alpinetin,7#ammonium glycyrrhizinate,9#honokiol,10#magnolol,11#costunolide,12#dehydrocostuslactone.The similar degree of the UPLC fingerprints of 15 batches of Houpowenzhong decoction was above 0.9.(3)A method for the determination of magnolol and honokiol in Magnolia officinalis,hesperidin in tangerine peel and alpinetin,pinocembrin,alnusenone and cardamomin in alpinia katsumadai was established.There exists a linear relationship for magnolol and honokiol in the range of 0.0048～0.028mg/ml and 0.00252～0.0252mg/ml respectively.The recovery rates are 100.61%and 99.42%,and the RSD are 2.63%and 2.91%.Hesperidin showed a linear relationship in the range of 0.01997～0.1997 mg/ml,with a recovery rate of 101.02%and an RSD of 2.11%.Alpinetin,pinocembrin,alnusenone and cardamomin have good linear relationships in the range of 0.00456～0.02785mg/ml,0.002775～0.013875mg/ml,0.000324～0.0012144mg/ml,0.002625～0.013125mg/ml.The recovery rates were 99.75%,96.52%,98.41%,and 97.45%,and the RSD were 3.98%,3.60%,8.75%,and 4.62%,respectively.Conclusions:In this thesis,based on the material benchmark research of Houpowenzhong decoction,the associative fingerprints of raw materials,decoction pieces,extracts,and lyophilized powders were established to realize the quality control of Houpowenzhong decoction and lay the foundation for clinical medication’s safety and effectiveness. |
