The Variety Of Tortoise Shell, Leech And Quality Study

The content of this paper includes the identification of the original a nimal, DNA bar coding, effective components or index component contents det ermination, fingerprint of H1RUDO and TESTUDINIS CAKAPAX ET PLASTRUM, which can evaluation these two herbs quality.Research purposes:this topic aims at traditional identification cannot describe the truth and quality of animal medicinal materials quickly, obje ctively and correctly, choosing HIRUDO and TESTEDIMS CARAPAX ET PLASTKTIM a s the instance, Making out the following research goal-Through the combination of DNA bar code, traditional identification and fingerprint, improve the identification method of HIRUDO and TESTUDINIS CA RAPAX ET PLASTRUM, which can basis for establishment a new, objective, obje ctive, digitized, standardized, quality standard of ani-mal medicinal mater ialsResearch methods:this topic used the following research methods:First, the traditional identification:including animal identificatio-n, character identification, microscopic identification and TLC iden-tificati on of animal medicinal materials, According to the results, preliminary jud ges its strainssecond, DNA barcode:DNA was extracted by kit method, and use univers-a1primer and Taq enzyme or TaqMIX enzyme to amplification, after se-quencin g, DNA bar code is obtained. Through, analysis of mutation si-tes and DNA b ar code genetic N-J tree, the leech and turtle’s strain is identified objec tively.Third, determination of effective or index components:according to t-h e reports, use hypoxanthine and L-hydroxyproline as standard subst-ance, d etermination hypoxanthine of HIRUDO and collagen of TliSTUDINI-S CARAPAX ET PLASTRUM. According to the2010edition of "Pharmaeopoe-ia of China", study the potency of thrombin of HIRUDO.Fourth, fingerprint:using HPLC determination method, according the rep orts to establish HPLC fingerprint. HPLC fingerprint drawing HIRUDO herbs, and compare and Study on its.The conclusion of the study:Using the traditional identification, ca-n make difference between the TESTUDINIS CARAPAX ET PLASTRUM’s origi-nal ani mal. But through the character, microscopic identification an-d TLC, It’s h ard to identification the a-nimal of HIRUDO and materia-ls. For these reaso ns, the author researches the DNA extraction, PCR amplification of leech, t urtles, and their materials, finally obtain-ed DNA bar code successfully, a ccording to the analyze of DNA bar co-des, the following conclusions can be drawn:the TESTUDINIS CARAPAX ET PLASTRUM, HIRUDO and its adulterants has a great many of variable sites. The genetic N-J tree shows that genuine and adulterants can be separated from the clear zone, so the DNA bar code tech nology can be identify the TESTUDINIS CARAPAX ET PLASTRUM, HIRUDO and its ad ultera-nts. This paper provides a new method for fast, accurate identificat,^ion of species of the medicinal materials. However, because the DNA bar co de only can do TESTUDINIS CARAPAX ET PLASTRUM, HIRUDO authentic-ity identif ication, its quality cannot distinguish. Therefore requir-es a combination of content determination, can be scientific, accura-te evaluation of TESTUD IMS CARAPAX ET PLASTRUM, HIRUDO quality.Using the "Pharmacopoeia" method, the extractives content of HIRUDO, TE STUDIN1S CARAPAX ET PLASTRUM were determined, the result shows that due to the effect of HIRUDO processing method, Whitmania pigra,Whitm-an, the large st amount of market share, has a greatly of difference of extract content.A nd the Chinese herbal pieces has low contents. Th-rough the measurement of the HIRUDO potency of thrombin, make a prel-iminary judgment of HIRUDO anti coagulant ability, it shows that ther-e is a great relationship between-coa gulation HIRUDO and varieties, varieties of blood-sucking HIRUDO anticoagul ant ability, not blood an-ticoagulant HIRUDO species ability.Through the determination of hypoxanthine, index components of HIRUDO, contents and collagen, effective components TESTUDINIS CARAPAX ET PL-ASTRUM. content, obtained the following conclusions:HIRUDO, from the measurement r esults can be seen, genuine of hypoxanthine content Whi-tmania pigra,Whitma n is highest, the lowest is Hirudo nipponica, Wh-itman. It’s in contrast to the results with thrombin. The anticoagul-ant components of HIRUDO has not been determined, therefore, the det-ermination of the composition of HIRUD0is the one side the quality of evaluation, market, mainstream varieties of HIRUDO, the index has c-ertain significance to the quality evaluation. TES TUDINIS CARAPAX ET PLASTRUM, the collagen content in genuine source is the highest, Mau-remys mutica has slightly lower content, common adulterants tr achemy-s scripta is lowest, the collagen content of Chinese herbal pieces o-f TESTUDINIS CARAPAX ET PLASTRUM is greatly reduced. Therefore, from the p erspective of content determination, the content of effective components wi th TESTUDINIS CARAPAX ET PLASTRUM is higher than its Chinese herbal pieces. In this study,HIREDO as example, research of10samples of different o rigin of the fingerprints, fingerprint shows the different origin of high imilarity, chemical composition of different origin contain roughly the sam e. But RSD of the share peak area is very large, so the common ingredient c ontent of samples difference is very big, it should be concerned with the c ultivation, breeding environment, processing method.To sum up, through the research the shortcomings of traditional evaluat ion of HIRUDO, TESTUDINIS CARAPAX ET PLASTRUM quality, explore the DNA bar code and combine four traditional identifications. Make a judgment of HIRED0, TESTUDINIS CARAPAX ET PLASTRUM quality comprehensively. Finally, this pr-oject makes a experimental basis of comprehensive quality of animal medicin e Pharmacopoeia of China2015edition.The research innovations:1, the project is in the traditional Chinese medicine under the guid-an ce of the theory, the molecular identification of medicinal mater-ials base, combined with the traditional character identification, microscopic identi fication, physical and chemical identification and content determination.2, for the first time on the TESTUDINIS CARAPAX ET PLASTRUM, HIRUDO, va riety, quality of original investigation and research system, HIRUDO, TESTU DINIS CARAPAX ET PLASTRUM quality evaluation.3, according to a series of requirements to establish the fingerprin-t of the HIRUDO, for example, establish the work accomplished the f-irst whol e HIRUDO fingerprint system.