Studies On S. Miltiorrhiza, S. Evansiana And S. Yunnanensis Based On Cytology, Identification, Medicine Components And Inorganic Elements

The genus Salvia Linn.(tribe Salvieae, Lamiaceae) represents an enormous andcosmopolitan of nearly 1050 species displaying a remarkable range of variation.Salvia has undergone marked species radiations in tropic and temperate zones, mainlydistributing in the coastal area of Mediterranean, central Asia, highlands of Mexicoand Andes mountains in South America. There are almost 78 species, 24 variations, 8formas in China, distributing in most areas especially the southwest regions. Amongthe 110 species of Salvia in China, 33 species (including variations and formas) havethe medical value, 26 of which could be used as the ingredients of traditional Chineseherb, such as S. evansiana and S. yunanensis etc..Salvia miltiorrhiza Bunge. is a perennial herb plant which has been used for along time in China. It is also called Xue Shen or Zidan Shen, which is the only sourceof Chinese herb Danshen according to "Pharmacopoeia of the People's Republic ofChina". As the well-established traditional Chinese herb, S. miltiorrhiza was firstdescribed in "Shin-nong Bonchokyeong" and was classified as top grade with theeffectiv treatment of coronary disease, heart angina and menoxenia appetite. Wild S.miltiorrhiza is often found near the places whose altitude is about 120～1300m, suchas the street of highlands, sloping fields and alongside valleys. In China, the mainproducing areas of S. miltiorrhiza are Sichuan, Henan, Shandong, Zhejiang, Jiangsu,Hubei, Anhui, Hebei, Shanxi and Hunan provinces. Especially in Sichuan province, S.miltiorrhiza has nearly one century history of being cultured and is famous for itsquality both in and aboard.In order to probe into the genetic diversity, microidentification characteristics,the content of medicine components and trace elements of S. miltiorrhiza, S. evansiana and S. yunnanensis, chromosomes number studies, character andmicroidentification analysis, medicine components and trace elements analysis werecarried out in this study. The main results are shown as follows:(1) The characteristics of S. railtiorrhiza from 6 provinces, S. evansiana and S.yunnanensis from Yunnan were observed and analyzed. Results indicated that thecharacteristics of character and microidentification of S. miltiorrhiza from 6 provinceswere the same as the descriptions of Radix Salvia Miltiorrhizae in Chinese medicineIdentification. In the traits of medicine character, S. evansiana had many branch rootswhich were like distorted braids, and its colour was bright red and sapidity was dense;S. yunnanensis had many intensive leaves streaks, and the root branches were smalllike fusiforms. In the characteristic of microidentification, the xylem of S. evansianaand S. yunnanensis were not gathered as sheaf, with fewer catheter cells to spread andwithout central fiber bundles. The results of the study could provide the genuineidentification characteristics of S. evansiana and S. yunnanensis and guide the drugsmarket to recognize the same herbs. And we also found that the cortexes of S.miltiorrhiza from Sichuan, Shandong and Henan provinces were much stronger thanother accessions, which might have something to do with the content of tanshinoneⅡA. The xylems of S. miltiorrhiza from Shandong, Henan, Hubei and Zhejiangprovinces were much stronger and had more catheter cells, which was presumed to berelated to the content of water-soluble components.(2) Accessions of S. Miltiorrhiza, S. evansiana and S. yunnanensis wereanalyzed by cell genetics technique. Results indicated that accessions of S.miltiorrhiza and S. yunnanensis had the same chromosome numbers 2n=8x=16; thechromosome number of S. evansiana was 2n=4x=32; in the cell of S. miltiorrhizafrom Zhejiang province, there was a chromosome with satellite. As the chromosomesare the small or micro-small ones, it was difficult to use them for karyotyping. Andsome nature tetraploid cells whose chromosome numbers were 2n=4x=32 wereobserved in S. miltiorrhiza from Sichuan and Shandong provinces. So we infered thatthe cell polyploidy phenomenon might be connected with the quality of crude drugs. (3) The contents of three medicine components TanshinoneⅡA, Salvianolicacid B, Protoctechual dehyde I were determined by High Performance LiquidChromat-ography (HPLC). Results indicated that the contents of TanshinoneⅡA inS.miltiorrhiza from Sichuan and Henan provinces were much higher, 4094μg/g and4268μg/g respectively; the contents of Salvianolic acid B in S. miltiorrhiza fromZhejiang and Sichuan provinces were much higher, 34.62 mg/g and 28.57 mg/grespectively; the contents of Protoctechual dehyde I in S. miltiorrhiza from Hubei andShandong provinces were much higher, 939.8μg/g and 897.8μg/g respectively. Thecontent of TanshinoneⅡA in S. evansiana was 2549μg/g which could satisfy therequest of "Pharmacopoeia of the People's Republic of China", and it could be usedas source herbs when TanshinoneⅡA was needed. The content of TanshinoneⅡA,Salvianolic acid B and Protoctechual dehyde I in S. yunnanensis were too low to beused as medicine herbs.(4) The contents of ten kinds of inorganic elements in accessions of S.miltiorrhiza, S. evansiana and S. yunnanensis were tested by method of Atomicabsorption spectroscopy (AAS). Results showed that in S. miltiorrhiza from Zhejiangprovince, the contents of trace elements Ca, Mg, Fe, Mo, Cu, Cd and Pb were thehighest (11.07 mg/g, 10.73 mg/g, 1.333 mg/g, 1.447μg/g, 27.03μg/g, 0.2610μg/gand 2.165μg/g respectively). So S. miltiorrhiza could be regarded as the herb with thestrong ability to absorb inorganic elements. In this accession the content of harmfulelement Cu exceeded the index of "Pharmacopoeia of the People's Republic of China".Meanwhile, the content of harmful elements Cd and Pb is much higher than someaccessions, thus S. miltiorrhiza from Zhejiang province is not agreeable to extend inthe herb medicine market. The contents of harmful elements Cu and Pb were alsomuch higher in S. yunnanensis(0.1850μg/g and 1.103μg/g respectively), so thisaccession is also not a good substitute for S. miltiorrhiza.