Pharmacognosy Study On The Tibetan NARDOSTACHYOS RADIX ET RHIZOMA

Nardostachyos Radix et Rhizoma was the dried root and rhizome of Nardostachys jatamansi DC.,collected by the calendar version of Chinese Pharmacopoeia.Clinically,it was the effect of "regulating qi and relieving pain,opening depression and xingpi,external use clearing damp detumescence".It was common medicine of Ayurveda and Unani of ancient India,and now it was common clinical medicine of Chinese medicine,Tibetan medicine,Mongolian medicine,Uighur medicine,Lisu medicine and Naxi medicine.NARDOSTACHYOS RADIX ET RHIZOMA was used as a medicine in Tibetan medicine for pangbei(bangbei).It was an essential medicine for the treatment of old febrile diseases,heat poisoning syndrome and plague.It was also one of the main raw materials of Tibetan incense.Its wild resources were mainly distributed in the Himalayas.In Chiana,the wild NARDOSTACHYOS RADIX ET RHIZOMA was mainly distributed in Sichuan,Tibet,Qinghai,Gansu and Yunnan.Due to the fact that the wild resources were the main source of medicinal materials of NARDOSTACHYOS RADIX ET RHIZOMA,its resoures was decreasing year by year and appeared to be endangered.At present,"the whole plant of N.jatamansi" replaces "the root and rhizome of N.jatamansi" in the market.It was researched by textual research on Tibetan medicine classics;medicinal plant resources investigation;identification of origin including the original plant,morphological,microscopic,DNA barcode etc.;multi-index screening of quality advantage and disadvantage of different habitats;different habitats and different plant parts of inorganic element content determination;volatile oil GC-MS chemical fingerprint.It was established comprehensive evaluation the quality of "the whole plant of N.jatamansi" and "the root and rhizome of N.jatamansi".It was expected to provide a scientific basis for the scientific and rational use of wild resources of the Tibetan NARDOSTACHYOS RADIX ET RHIZOMA and whether the new edition of Chinese pharmacopoeia could replace the root and rhizome of N.jatamansi with "whole plant of N.jatamansi".The main research contents were as follows:Research on the name "Gansongxiang",which was frequently recorded in Tibetan medicine and modern Chinese medicine literature mainly includes N.jatamansi(D.Don)DC.,N.chinensis Batal.or N.grandiflora DC.(abroad),by combining with 31 samples collected from Tibetan areas in four provinces of China,the identification research was the original plant,the test results were as follows:(1)after the founding of new China,the 1953 edition of the Chinese pharmacopoeia was not recorded N.jatamansi.In 1957,a pharmacognosy study on N.jatamansi,it was considered that the original plant of N.jatamansi was 1 species of N.chinensis Batal..Since the Chinese pharmacopoeia(1963),only 1 species of N.chinensis Batal.was recorded.Chinese pharmacopoeia,1977,1990,1995,2000 and 2005 edition,all contain two species of N.jatamansi,N.chinensis Batal and N.jatamansi DC.,the 1985 edition of Chinese pharmacopoeia were not recorded of N.jatamansi.The 2010 and 2015 versions of Chinese pharmacopoeia were derived from 1 species of N.jatamansi DC.However,the Chinese name of N.jatamansi in the previous version of Chinese pharmacopoeia was used the Chinese name of "N.chinensis Batal." and the Latin name of N.jatamansi(D.Don)DC..(2)N.jatamansi(D.Don)DC.,N.chinensis Batal.,N.jatamansi(N.chinensis)and N.grandiflora DC.were actually a polymorphic variety of the same species,which was suggested be merged by our research group.Therefore,the source of the medicinal material Gansong recorded in the Chinese pharmacopoeia in 2010 and 2015 was one species,namely”N.jatamansi DC.”.Our Research group suggested that the original plant was N.jatamansi DC.and its variant of N.jatamansi(D.Don)DC.f.plena Batal.To strengthen the protection of endangered species of N.grandiflora(reference CITES 2007,2013),it was conducted the field collection and trade status survey of of N.jatamansi [N.jatamansi(D.Don)DC.,N.chinensis Bat.] in Sichuan,Qinghai,Gansu and Tibet.The research results showed that:(1)It was an important plateau plant resource with medicinal value,ecological value and economic value.(2)N.grandiflora DC.was listed in CITES2007,2013 appendix II,but the Chinese name used N.jatamansi(D.Don)DC.The Latin equivalent was N.grandiflora DC..It was suggested that wild resources should be modified for the distribution of wild resources in foreign countries N.grandiflora DC.and was listed in CITES.(3)N.jatamansi was an important raw material of Tibetan incense and Buchang Wenxin Granules,Songbuli oral liquid,Shangtongning capsule and other proprietary medicine,Tibetan incense and daily chemicals.However,as it was still completely dependent on wild resources,with the actual production of cypress demand increased year by year,the demand for cypress was increasing.In Tibetan areas in Sichuan,Tibet,Qinghai,Gansu,Yunnan and other provinces,a large number of Tibetan temples and Tibetan family buddhist sacrificial activities,a small number of household exorcising evil effects,on behalf of the essence of Tibetan culture,"Tibetan incense" has been prohibited to add raw materials of N.jatamansi plants.It was learned from traditional patent medicine enterprises and Tibetan incense factories that the regeneration of wild resources of N.jatamansi(D.Don)DC.was very slow,which could not meet the needs of enterprises.Tibetan incense factories and export was in a state of gradual decline at present.(4)Due to the harsh requirements on the ecological environment,the artificial planting was not yet formed a scale,and the wild reserves were gradually reduced,the market was basically "the whole plant of N.jatamansi" to replace "the root and rhizome of N.jatamansi(D.Don)DC.".(5)The wild N.jatamansi(D.Don)DC.resources were mainly distributed in Ruoergai county,Aba Tibetan and Qiang autonomous prefecture,Sichuan province,Baiyu township,Jiuzhi county,Qinghai province,and Maqu county,gansu province.(6)N.jatamansi i(D.Don)DC.was artificially cultivated in the qinghai-tibet base of southwest university for nationalities(Qiongxi town,Hongyuan county,Aba prefecture,Sichuan province)of 30 mu,and the agricultural science research institute of ganzi prefecture of 5 mu.However,it has not been able to provide seeds or seedlings or medicinal materials to the market.In order to further identification of origin of NARDOSTACHYOS RADIX ET RHIZOMA,the identification of the original plant,character,tissue structure and powder were studied.The test results showed that:(1)The traditional identification method could be distinguished,but the results still need to be resampled in the summer of 2019 for repeated verification.(2)After the ITS sequence was amplified from ITS,it was identified that all the samples collected from the producing areas were N.jatamansi DC..It was preliminarily considered that the Chinese pharmacopoeia of 2015 edition contains a large possibility of N.jatamansi DC.Variation was N.chinensis Batal..This result was the same as the literature research and the original plant identification results of the samples collected in the main producing areas of N.jatamansi DC."the origin of N.jatamansi DC.listed in the Chinese pharmacopoeia in 2010 and 2015 was one species,namely N.jatamansi DC..Research group research that the original plant of N.jatamansi DC.and its variant N.jatamansi(D.Don)DC.f.plena Batal.almost the same.(3)Rbc L sequences were not completely consistent with ITS sequences and morphological classification and identification results,and the relationship between the sequence information of cyperus manshuriensis and ITS variation and altitude and origin had not been found,In combination with multi-sequence analysis,Our research group was planning to establish the molecular fingerprint markers of N.chinensis Bat.and N.jatamansi(D.Don)DC.,further identification of kinship between N.chinensis Bat.and N.jatamansi(D.Don)DC..Based on the premise of wild resources protection of NARDOSTACHYOS RADIX ET RHIZOMA,whether NARDOSTACHYOS HERBA(NH)could be equal to NARDOSTACHYOS RADIX ET RHIZOMA(NRR)specified in pharmacopeia for medical use or not,and to discuss the scientificity of using NH as the commodity specifications in market circulation.The gray correlation,regression analysis and PCA and HCA analysis were used to analysis the data,including the root length,plant height,dry weight,content of volatile oil,water-soluble extract,content of nardosine,moisture,total ash,acid-insoluble ash,etc.,in order to comprehensively evaluate the quality of NH and NRR.The quality of NH and NRR were mainly correlated with the content of volatile oil and water soluble leach of NRR.Two variables corresponding to the dependent variable,aqueous extract and volatile oil,of total eight variables had significant influences between NH and NRR(P < 0.01).The multidimensional statistical analysis of medicinal material quality illustrated a obvious difference between NH and NRR.It was conformed once again the NRR was to be the best for medical use,which was the same indexed regulations in different version of Chinese Pharmacopoeia.The NH was not absolutely substituted for NRR.To provide experimental basis of “the whole plant of Nardostachs jatamansi DC.” for market being scientific and reasonable.Simultaneous determination of Inorganic Elements of different parts and different habitats by Wet Digestion-ICP-OES.The results were analyzed by correlation analysis,principal component analysis,cluster analysis.The test results showed that: It was a significant correlation among the elements.The principal component analysis were adopted to determine the elements of the above ground parts was Pb,Se,As,Si,Mo,Co,Ni,Cd,Cr,Tl,Na,Sn.The underground part was Co,Cu,Fe,Mn,Mo,Al,Ba,Ca and Mg.The above ground and underground results were well classified by cluster analysis.The above ground and underground parts of NARDOSTACHYOS RADIX ET RHIZOMA at different altitudes were analyzed to determine the optimum growth altitude.The experiment could provide some reference for quality control and resource utilization of NARDOSTACHYOS RADIX ET RHIZOMA.To establish the fingerprint of volatile oil components about N.jatamansi DC..in different regions,and to comprehensively evaluate the quality consistency and difference of the medicinal materials by the Total Statistical Moment Method combined with the Principal Component Analysis and Cluster Analysis.It provides a reference for the quality control of N.jatamansi DC..GC-MS method was adopted to establish the 15 origin N.jatamansi DC..Fingerprint SIMCA-P 13.0 software was used to evaluate the similarity by the Principal Component Analysis and cluster analysis,principal component contribution rate,clustering analysis of distribution.There were 6 components in 15 producingareas.Respectively,?-Patchoulene,?-Elemene,(1a R)-1a?,2,3,3a,4,5,6,7b?-8hydrogens-1,1,3a?,7-tetramethy-1H-cyclopropane[a]naphthalene,Aristolene,?-Patchoulene,?-bulnesene.Besides S13 samples,respectively,it was similar to the rest of the similarity,the chromatographic fingerprint similarity evaluation system of Chinese medicines,obtained similarity above 0.708,the total counts moment method,similarity above 0.812,PCA analysis and HCA analysis samples were divided into four categories.Besides S13 samples,respectively,it was similar to the rest of the similarity,the chromatographic fingerprint similarity evaluation system of Chinese medicines,obtained similarity above 0.708,the total counts moment method,similarity above 0.812,PCA analysis and HCA analysis samples were divided into four categories.Among them,for a class of Sichuan(S1,S5),Qinghai(S6,S8,S9),Gansu(S11);Qinghai(S7),Gansu(S10);Sichuan(S2,S3,S4);Gansu(S12),Tibet(S14,s15).Sample 13 was different greatly from the other 14 samples.GC-MS fingerprint was established,and the total statistical moment method,combined with principal component analysis and cluster analysis method was adopted comprehensive evaluation of the consistency and difference of NARDOSTACHYOS RADIX ET RHIZOMA to provide certain reference for the quality evaluation.