A Comprehensive Evaluation Study On The Quality Of Some Medicinal Materials In Chicory

BackgroundThis research was part of the research of NSFC(NO.81073028)and SRFDP(NO.20130013120001)which was supported by Beijing University of Traditional Chinese Medicine(2013-JYB22-XS-086).Herba chicori is well known in Uighur folk medicine as a cholagogic and diuretic agent to improve the appetite,increase digestion and treat liver diseases,etc.The function of hypoglycemic,hypolipidemic,lower uric acid,improve immune function and regulate intestinal flora of chicory has been proved by modern research.For the mentioned reasons,chicory has a bright prospect in future.However,there is a lot of fake and bad quality chicory available in the market,which will affect its safety and effectiveness.In the Pharmacopoeia of the People's Republic of China,the quality of Cichorium intybus L.and C.glandulosum Boiss.et Huet.evaluated only by its characters,microscopic characters and few chemical characters which is hardly to identify its species and evaluate its quality.The development of the methods for evaluation the quality of Herba chicori is very important to ensure its safety and effectiveness.The purpose of this experiment is to establish the methods to evaluate the quality of stem of Cichorium intybus L.and C.glandulosum Boiss.et Huet..Research ContentThis study includes theoretical and experimental research in two parts.Theoretical research1.Review on the quality evaluation methods of Herba chicoriA systematic review on the evaluation methods of Herba chicori,including microscopic identification,TLC,determination of the active ingredient,general infrared spectroscopy,HPLC fingerprint,determination of pesticide residues and heavy metals.2.Review on the quality evaluation methods of TCMA systematic review on the evaluation methods of chicory,including the ordinary traditional evaluation ways,the methods based on the determination of active ingredient or content ratio,the methods based on HPLC fingerprint,the methods based bioassay,the methods based TLC-bioautography.This review provided a good foundation for the choosing of effective ways to evaluate the quality of chicory.3.Review on the use of HPLC fingerprint and rDNA-ITS on the quality evaluation of TCMA systematic review on use of HPLC fingerprint and rDNA-ITS in the quality evaluation of TCM.This review provided a good foundation for further research on the evaluation of the quality of chicory.Experimental ResearchThis study built the methods to evaluate the quality of Herba chicori by identification way and quality evaluation way.The identification side include Characters identification,microscopic identification,TLC,rDNA-ITS.The quality evaluation way include HPLC fingerprint,determination the Chicory acid,Chlorogenic Acid and polysaccharides contents of chicory.This research includes three segments:Part one:The identification of Herba chicoriExperiment 1:Traditional ways to identify Herba chicoriThe ordinary traditional way to identify Herba chicori includes Characters identification,microscopic identification,TLC.The three ways can identify raw herbs of Herba chicori effectively,but in clinical practice the use of Herba chicori is pieces,whether the three ways can identify Herba chicori well?The purpose of this experiment is to answer this question.Experiment 2:The study of ITS sequence of Herba chicoriThis experiment analyzed the ribosomal DNA internal transcribed spacer(rDNA-ITS)sequence of Cichorium intybus and C.glandulosum from different habitats,and to provide DNA molecular marker for the identification of Herba chicori.Part two:The establish of HPLC Fingerprint of Herba chicoriExperiment 3:The establish of HPLC Fingerprint of Herba chicoriIn this experiment,the software 'Similarity Evaluation System for Chromatographic Fingerprint of TCMs'(Version 2004A)was employed to generate the mean chromatogram and carry out the similarity analysis of the samples.Cluster analysis was adopted in combination with principal component analysis to study seventeen Herba chicori's seventeen characteristic common peaks and to distinguish the two chicory resource.Part three:Study on the content of three chemical composition of Herba chicoriExperiment 4:Study on the content of Chicory acid,Chlorogenic Acid contents of Herba chicoriThis experiment is to determine the Chicory acid and Chlorogenic Acid of Cichorium intybus L.and C glandulosum Boiss.et Huet.from different regions and provide the scientific basis for its application.The Chicory acid and Chlorogenic Acid contents were determined all by HPLC method.Experiment 5:Study on the content of polysaccharides contents of Herba chicoriThis experiment is to determine polysaccharides contents of Herba chicori from 15 different regions and provide the scientific basis for its application.Results1 Characters identification,microscopic identification,TLC can be used to indentify Herba chicori.But stability of the Characters identification and microscopic identification is bad.The TLC method can't indentify Cichorium intybus L.from C.glandulosum Boiss.et Huet.for the chemistry similarity.2 The intraspecies identity of ITS sequence was above 99.2%in C.intybus,and that in C.glandulosum was above 99.8%,while the interspecies identity of ITS sequence was below 99.2%.There were various specific information sites in the ITS sequences of the two kinds of Herba chicori.3 The fingerprint of Cichorium intybus L.and C.glandulosum Boiss.et Huet.were set up,and the range of similarity for seventeen chicory 0.847?0.988.The differenee among chromatographic fingerprints of Herba chicori samples between the two different varieties was identified by cluster analysis and principal component analysis.4 The contents of Chicory acid and Chlorogenic Acid of Herba chicori from various cultivation locations varied from 2.370?6.217 mg/g and 0.0367?1.2986 mg/g respectively.5.The contents of polysaccharides of Herba chicori from various cultivation locations varied from 2.593?7.075 mg/g.Conclusion1 The ordinary traditional evaluation ways is convenient,but is not stable.The ITS sequence is an available molecular marker for the identification of C.glandulosum and C.intybus.The ordinary way should combine with ITS sequence to identify Herba chicori for both of them have its own advantage and disadvantage.2 HPLC fingerprint method can be used to evaluate the quality of Cichorium intybus L.and identify Cichorium intybus L.from C.glandulosum Boiss.et Huet.conveniently.The similarity of tested herb and Herba chicori sample should past 0.85,and they both should have 17 common peaks.3 Chicory acid and Chlorogenic acid were important content of Herba chicory,and the content of Chicory acid and Chlorogenic acid can be used to evaluate the quality of Herba chicori.The content of Chicory acid and Chlorogenic acid should past of 2mg/g and 0.2mg/g of Cichorium intybus L.respectively.The content of polysaccharides in the stem of Herba chicori was stable from different regions,and its can be also used to evaluate the quality of Herba chicori.The content of polysaccharides should past of 2mg/g.