The Research Of Extraction And Quality Control Technique In The Separation And Analysis Of Active Components Of Portulaca Oleracea L.

The quality of Chinese herds has an important role on the effectiveness of prevention and treatment of disease and closely related to people's health and safety. In order to ensure the safety, rational and effective of drugs the testing should be rigorous operated and comprehensive quality of traditional Chinese medicine should be controlled in the process of preparation, production, storage, supply and clinical research of traditional Chinese medicine. It is the main issues and important tasks of the modernization of Chinese medicine that modern analysis methods and techniques be used in Chinese medicine research and production process to improve the quality of Chinese medicine and speed up the modernization and internationalization of Chinese medicine. The noval types of sample pretreatment techniques (microwave-assisted extraction approaches, molecular imprinting technique and solid-phase microextraction) have been used for extraction of flavonoids from Portulaca oleracea L. In this work, the parameters which influence the extraction efficiencies in each technique had been investigated. Additionally, the quality control standard of Portulaca oleracea L. was established by the techniques of chromatographic and spectroscopic techniques, which was used to control the quality of Portulaca oleracea L. And then sample pre-treatment methods of the flavonoids, such as kaempferol, from Portulaca oleracea L. was studied, which can provide reference informations for the rational use of Portulaca oleracea L. The concrete contents are as follows:1. Analysis of Flavonoids in Portulaca oleracea L. by UV-Vis Spectrophotometry with Comparative Study on Different Extraction TechnologiesUsing Portulaca oleracea L. as the research object and the extraction method of total flavonoids in Portulaca oleracea L. was studied. Five extraction technologies of flavonoids from Portulaca oleracea L. were investigated and compared, including microwave-assisted extraction, ultrasonic extraction, reflux extraction, Soxhlet extraction, and marinated extraction. The conditions of microwave assisted extraction were optimized by single factor experiments and orthogonal test, and the extraction solvent, extraction time, extraction temperature and extraction of solid to liquid ratio was investigated. Quantification was performed by means of UV-Vis spectrophotometry with chromogenic system of NaN02-Al (NO3)3-NaOH. In alkaline conditions the chromogenic system has a characteristic absorption in the visible region and could be determined. The results showed that microwave assisted extraction were most suitable for the extraction of flavonoids from Portulaca oleracea L. because of its high effect and short extraction time. The found optimum extraction conditions were that the ethanol concentration was 70%(v/v), solid-liquid ratio was 1:50, extracting temperature was 50℃and irradiation time was 9 min. Under the optimum conditions, the linear regression equation was A=-0.00171+11.58482C, the calibration curve for the analyte was linear with the correlation coefficients was greater than 0.9999. The average recovery was 102.6%, and its RSD was 1.13%(n=5). Eight types of Portulaca oleracea L. according to different habits were investigated. The total content of flavonoids was 7.16,7.10,9.38,6.82, 6.78,11.36,5.12, and 1.76 mg/g, respectively.2. Identification of Portulaca oleracea L. from different sources using GC/MS and FT-IR spectroscopyA fingerprinting approach was developed by means of gas chromatography/mass spectrometry (GC/MS) and IR spectroscopy for quality control of Portulaca oleracea L., and the similarity evaluation and hierarchical cluster analysis were performed to evaluate the similarity and variation of these samples. Major components of volatile oil and characteristic peaks in the common pattern were identified. The results show that the information of the compounds and the information of the relative content of the compounds could be obtained through chromatography mass spectrometry analysis and The IR spectrum shows a total overlap of each absorption spectrum of all components. The cluster analysis results of GC/MS and IR spectra were similar. GC-MS fingerprint can be used for the rapid separation and identification of major components and the second derivative spectrum can enhance the apparent resolution of IR spectrum.3. Development and Characterization of Molecular Imprinted Polymer for the Selective Detection of Kaempferol in Traditional Chinese MedicinesMicrowave heating was applied to the preparation of kaempferol molecular imprinted polymer microspheres, the term of the polymerization was dramatically shortened by using microwave heating and the results of morphology observation, static absorption performance and selectivity performance of the MIP microspheres were all superior to the MIPs prepared by conventional heating. The MIP microsphere were demonstrated with a narrow diameter distribution (6-9μm) and with a spherical shape and the imprinting efficiency of the MIP microspheres prepared by microwave heating was 5.0 and by conventional heating was 4.2. Based on the imprinting effect, the MIPs prepared by microwave was used as the sorbent of solid phase extraction and the properties of the resultant extraction cartridge shows a good extraction performance of kaempferol. The MIPs coupled with solid phase extraction was used for extracting kaempferol in Portulaca oleracea L. and Alpinia officinarum, the recoveries were 90.2% and 88.0% and RSD were 1.21% and 1.18%. The results indicated that the MIPs can be favorably used for the extraction of the kaempferol in traditional Chinese medicines.