Studies on Extraction and Purification of Flavonois and Saponins from Cyclocarya Paliurus
|School||Hunan Agricultural University|
|Keywords||Cyclocarya paliurus Flavonoids Saponins Extraction and purification|
Cyclocarya paliurus (Batal.) is one of the unique species, and classified as being protected endangered species in China. Its roots, branches, leaves all can be used as medicine that have been confirmed to lower the blood glucose and blood pressure, weight loss, anti-tumor, anti-aging, anti-allergic, diminish heat, enhance human immunity, promote metabolism, such as a wide range of effectiveness. Studies have shown that physical activity and these flavonoids, triterpenes and polysaccharides in Cyclocarya paliurus, such as closely related to the active ingredient. At present, research is focused on the functions and pharmacological activity of the isolation and identification of Cyclocarya paliurus, the active substances of its extraction, purification was not carried out systematic research, especially for extracting and purifying flavonoids and saponins synchronously there was no reported study. In this paper,these researches have been done, including the detect method of flavonoids and saponins content, the technics of extraction and purification synchronously,and comparative study for the main component of Cyclocarya paliurus between raw materials and made tea, which is provided a theoretical basis for the active ingredient and its effective use of Cyclocarya paliurus. The main contents and research results are as following:1、Establishing the determination method of total flavonoids and saponins in Cyclocarya paliurus by double-wavelength spectrophotometryDetermination of total flavonoids in the use of AlCl3, choosing 410 nm as detection wavelength and 470 nm as reference wavelength, establishing regression equation with the absorbance values ofΔA and concentration C (mg/ml), and C= 0.8567ΔA+0.004; R2= 0.9991. The results have shown that flavonoids in the method of linear correlation, stability, reproducibility, accuracy and degree of recovery are better. The method can be used for detecting total flavonoids in Cyclocarya paliurus.Determination of total saponins in the use of vanillin-glacial acetic acid perchloric acid, choosing 555 nm as detection wavelength and 590 nm as reference wavelength, establishing regression equation with the absorbance values ofΔA and concentration C (mg/ml), C= 10.449ΔA+0.0059; R2= 0.9979. Studies have shown that the method for total saponins quantitative detection of the initial compounds.2、The extraction conditions were optimized and the best technics parameters were obtained.In the extraction experiment, EtOH, a safe solvent without poison was used. The optimum conditions for the extraction were determined, which were 70% of alcohol concentration,30 times the volume of EtOH consumption, at the bath temperature of 90℃,time of 4 h extracted twice, The extract ratio under the conditions of flavonoids up to 3.055%, rate of saponins extracted 4.791%.3、The conditions of purification by macroporous resin were optimized.In the purification experiment, the X-5, D101,NKA-9, AB-8 macroporous resin were compared to the absorption and desorption efficiency. Ultimately determine the X-5 resin purification conditions are as follows:Cyclocarya paliurus extract concentration of 3.00～5.00 mg/mL with a flow rate 2.0 BV/h followed by washing the sample to the outflow of liquid and transparent, and then 4BV,80% EtOH, in order to 3.0 BV/h flow rate of the synchronous-eluting flavonoids and saponins. The elution rate under the purification conditions of the flavonoids and saponins respectively is 98.806% and 90.072%.4、Comparing of total flavonoids and saponins in Cyclocarya paliurus between raw materials and made tea.The results showed that the same season in the region of the mature leaves flavonoids and saponins content of all test samples for the highest levels; same processing tenderness compared to wild Cyclocarya paliurus two substances were all higher than artificial cultivation Cyclocarya paliurus of content; the same season of different artificial cultivation of the fresh leaves of child-bearing age compared to both its content was reduced after the first increase in the trend.