Researches On Systematic Identification And Quality Assessment Of Rubia In Anhui Province

Objectives:1. Investigating Rubia plants in Anhui province to get the local resources of the vine of R.cordifolia L. for Hefei Huarun Shenlu Drugs Ltd., Erxieting products.2. Doing the identification research on three kinds of Rubia plants in Anhui province,and establishing the systematic model with origin identification, macroscopic identification, physicochemical identification, microscopic identification and DNA barcoding identification. Evaluating the quality of medicine herbs of Rubiae quantitatively to establish the qualitative assessment system of medicine herbs. In this way, scientific reference can be provided for the classification, identification and the development and utilization of Rubia plants.Methods:1.The species as well as the distribution of Rubia plants in Anhui province are known by field investigation, sample collecting and specimen identification.2.To identify three kinds of Rubia plants systematically based on plant morphology,pharmacognostical characters,TLC identification,microscopic identification and DNA barcoding identification.3.To study the histochemical localization of anthraquinones of root in R.cordifolia L.on the basis of chromogenic agent chromogenic reaction principle.4.To determine the contents of Purpurin and Mollugin in the root and rhizoma of Rubia plants in Anhui province, medicine herbs of Rubiae on sale and the phloem and xylem of R.cordifolia L.Results:1.Three species of Rubia plants distribute widely in Anhui province,namely,R.cordifolia L.,R. argyi(Lévl.et Vant) Hara ex L.A.Lauener et D.K.Ferguson and R.alata Roxb. R.cordifolia mainly distributes in northern Anhui plain and Jianghuaihilly region. The Dabie Mountain, the river hilly region and mountain area in southern Anhui own the R. argyi resources. R. alata mainly distribute in Dabie Mountain and the river hilly region.2.The three kinds of plants have differences from plant morphology,pharmacognostical characters,TLC identification,microscopic identification,etc..The leaf of R.cordifolia is bigger,and the other is smaller.The shape of R.alata is lanceolate. Ripening fruit of R.cordifolia is nacarat, and its peel and seed separate easily. The xylem of R.cordifolia accounted for the entire root diameter ratio is about2/3,R. alata is 1/3 and R. argyi is 1/2,the ratio from large to small:R.cordifolia >R.argyi >R. alata. The conclusion of TLC identification is that acrial part of Rubia plants content flavonoids,root and rhizoma content Mollugin. When comes to the characteristics of the powder, R.cordifolia have two kinds of nonglandular, one is branched nonglandular.The micro-macroscopical observation results show the shape of the two kinds of nonglandular on the stem of R.cordifolia, one is olecranon bending,the other is hook. The success rates of 23 samples for DNA extraction, PCR and double chains measurement are ITS78.9%(15),psb A-trn H100%(19).The DNA and NJ phylogenetic tree combined from the ITS and psb A-trn H can directly reveal the genetic relationship among Rubia plants.The combined sequences can identify Rubia plants and the source of medicine herbs of Rubiae.3.The research on the histochemical localization of anthraquinones shows that anthraquinones accumulated mainly in the phloem of the root of R.cordifolia L.4.Two index components of content determination of Mollugin and Purpurin in medicine herbs of Rubiae by HPLC shows that chromatographic contions is mature,the shape of chromatographic peak is good,theoretical plate number,tailing facter and separation meet the requirement. Mollugin within the range of 0.0010 to0.216 mg/ml and Purpurin within the range of 0.0016 to 0.2005 mg/ml had good linear relationship with the peak area.The content of two index components in the root and rhizoma of R.cordifolia and R.alata are consistent with ChinesePharmacopoeia.Determination of index components of medicine herbs of Rubiae on sales,the samples got from Shanxi province and Hebei province can’t meet the requirements.Qualified samples have large difference on the content of Mollugin,the quality of medicine herbs of Rubiae is spotty.The content of Mollugin and Purpurin in phloem is higher than xylem in the root of R.cordifolia L.Conclusions:1.The local resources of the vine of R.cordifolia L. is found in Anhui province by field investigation and reduce the production cost of the enterprise.2.Origin identification,macroscopic identification, physicochemical identification,microscopic identification and DNA barcoding identification can be used for systematic identification of Rubia plants in Anhui province as well as providing a new method for accurate,rapid identification of Rubia plants. However, it also provides new ideas for Hefei Huarun Shenlu Drugs Ltd,. on making enterprise quality standards of Erxieting products.3.The index components of medicine herbs of Rubiae were determined by HPLC.The determination results show that the index components of the root of R.alata is also consistent with Chinese Pharmacopoeia description. In the future, it can be used as a alternative one to the use of medicine herbs of Rubiaeon the basis of its research of pharmacodynamics and recommended. Medicine herbs of Rubiae in circulation on sale may face serious quality problems, for examples,the source of medicinal material is not unified, the roots of same genus sales as medicine. Anthraquinones mainly accumulated in the roots of R.cordifolia L. of phloem. As a result, it provides the basis for the development and utilization of anthraquinones in R.cordifolia L.