Quality Evaluation Of Local Medicinal Material Of Yantai Bupleuri In Shandong

Bupleurum, which is commonly and large amount used herbs in Chinese medicine, has a long history in application. It has function of repaying hot, and normally used for colds, fever, chills and fever, malaria, Qi stagnation in liver, sternocostal pain, prolapse, uterine shedding, irregular menstruation and so on. The formal recorded Bupleurum in Chinese Pharmacopoeia are Bupleurum DC. or dried roots of Bupleurum scorzonerifilium Wild. Bupleurum is the main content of Dachaihu Soup, Xiaochaihu Soup, Chaihuguizhi Soup. Modern pharmacological studies confirmed it owns antiviral, anti-inflammatory, antipyretic and analgesic, liver and gallbladder, anticancer role and medicines, such as Bupleurum injection, oral Bupleurum, Bupleurum pills, made mainly by Bupleurum have a wide demand in clinical application. Thus as natural resources decrease, problem of shortage is more and more prominent. In China, there are42kinds of Bupleurum including17varieties,7variants, mainly growing in Liaoning, Gansu, Hebei, Henan, Shandong, Hubei, Jiangsu, Sichuan, Anhui and other places. Many regional Bupleurum are used as formal local medicinal Bupleurum all over the country or sold as formal Bupleurum. In addition to that, medicinal parts of these plants are not the same. For traditional Bupleurum, we use roots however whole plants or aerial parts are often used in some regions especially in south China. Some Bupleurum are whole-plant used in case of wasting. Now, because of the unsteady and complex situations in using Bupleurum, many pharmacists are studying its chemical constitutes and pharmacological effects, hoping to find alternation in some degree for the shortage of Bupleurum resource. Thus it is important to evaluate the quality of local herbs and meanwhile this evaluation will bring theoretical basis when expanding the introduction and cultivation in next steps.Yantai Bupleurum (BupLeururn chinense DC. f.vanheurckii(Muell.-Arg.)Shan et Y.Li) is specific to Shandong Province. It is a variant of Bupleurum DC. which difference appears only in edge color and shape of small bracts. It spreds on200-950elevation of meadows in Yashan, Aishan, Kunyushan etc. and local people often use it as genuine Bupleurum. Its leaves and stems are applied to make Chaitong Pill for children’s pneumonia for years, which indicates the whole plant owns medicinal functions. Now, there are no evaluation about Yantai Bupleurum. Therefore in this article we performed four experimental study and offered theoretical basis in its development.1. Pharmacognosy. This study was to carry out identification of original Yantai Bupleurum to make clear of its species. Meanwhile, we collected samples to perform characteristic and quality evaluation including traits, microscopic and chemical identification. The identification indicated that Yantai Bupleurum has the common traits of Umbelliferae. Under microscope, we could clearly observe its cork cells, thin-walled cells, oil cells, threaded conduit and fiber structure. Chemical identification showed it has triterpenoid saponins, flavonoids and volatile oil. Tlc identification showed that Yantai Bupleurum has saikosaponin-a and saikosaponin-d compared with genuine reference.2.Volatile oil and its antimicrobial activity. Objective:To study the components of the enssential oil from the aerial part of Bupleurum chinense DC. f. vanheurckii (Muell.-Arg.) Shan et Y.Li and its antimicrobial activities. Methods:The distillation of water vapor was applied to extract the enssential oil from Bupleurum chinense DC. f. vanheurckii (Muell.-Arg.) Shan et Y.Li.Then, GC-MS was used to analyse the components of it. The antimicrobial activities were investigated by plate diffuse and the minimum inhibitory concentration (MIC) was determined by plate spread. Results:The30components were successfully identified from the extracts of enssential oil Meanwhile, the essential oil has a broad-spectrum antimicrobial activities. Conclusion:The mainly components of the essential oil from Bupleurum chinense DC. f. vanheurckii (Muell.-Arg.) Shan et Y.Li are terpenes and low molecular aliphatic hydrocarbon compounds. And the antimicrobial intensity to the tested strains is:Staphylococus aureus> Penicillium citrinum> Escherichia coil> Staphylococcus albus> Bacillus subtilis> Aspergillus niger> Aspergillus flavus.3. Determination of flavonoids content. Objective:We used HPLC to determine rutin and quercetin extracted from whole Yantai Bupleurum plant. Methods: Agilent C18(250mm×4.6mm,5μm) column, mobile phase:acetonitrile-water (containing0.1%phosphoric acid), gradient elution method, flow rate:1.0mL/min; detection wavelength:360nm. Results:The linear range of rutin and quercetin are within10～100μg/mL, r=0.9999and0.9997, respectively. Average recoveries were100.47%(RSD1.34%) and99.50%(RSD0.41%). Conclusion:The method is simple, rapid, accurate and suitable for content determination of rutin and quercetin extract from Yantai Bupleurum whole plant.4. Determination of Sakosaponins. Objective:To provide scientific basis for evaluating the quality of Yantai Bupleurum through establishing HPLC method to determine and study the content of saikosaponins a, d in it. Methods:Thermo ODS-2HYPERIL(250×4.6mm,5μm)column was used. Mobile phase was acetonitrile-water with gradient elution at a flow rate of1.0mL·min-1. Column temperature was equal to room temperature and the detection wave length was210nm. Results:Methodology examination indicate that the linear relation between peak area integral value and saikosaponins a, d sample were good (ra,d=0.9999). RSD in precision experiment was0.67%and0.75%respectively. Meanwhile the respective recovery coefficient was99.4%and98.7%. There is only a little difference in content of saikosaponins a, d that exist in Yantai Bupleurum from different origins. The total content is between0.772%and1.079%. Conclusion:This method, which is stable, simple and with good reproducibility, can be used for the quality control of Yantai Bupleurum. Saikosaponins a, d content in Yantai Bupleurum is found higher than the standard content of which in Bupleurum Chinense DC. according to Chinese Pharmacopoeia, which provides laboratory foundation for the application of Yantai Bupleurum.The innovation of this paper:(1). For the first time carrying out pharmacognosy experiments in Yantai Bupleurum.(2). For the first time carrying out chemical composition of determination in Yantai Bupleurum volatile oils, flavonoids and saponins.(3). Initially developed quality standards of Yantai Bupleurum, provided a theoretical basis for its exploitation.