| Cedrus deodara is the general term of Pinaceae plants, traditional Chinese medicine thinks that Cedrus deodara has the effects of collaterals activating, winddispelling,stop bleeding,promoting blood circulation,detumescence and promoting granulation.Modern pharmacological studies have shown that Cedrus deodara has antioxidant, anti-tumor,anti-browning, antiviral and other pharmacologically active. Cedrus deodara are widely distributed in our country, and pine needles is rich in resources.In order to develop and utilization the pine needles of Cedrus deodara,this thesis aimed at made a system research on antioxidant activity of total polyphenol from pine needles of Cedrus deodara.The main contentand research results are follows:1.Simultaneous determination of gallic acid,protocatechuic acid and catechin in pine needles of Cedrus deodara by HPLC.Agilent TC-C18 column(250 mm×4.6 mm, 5 μm) was adopted. The mobile phase was methanol(A)- 0.2% phosphoric acid(B) with the gradient elution(0-8 min, 10%Aâ†'10%A; 8-45 min, 10%Aâ†'30%A) at the flow rate of 1.0 m L·min-1.The column temperature was 25 ℃ and the detection wavelength was 280 nm.The results show that calibration curves were found to be linear in the ranges of 0.4~4.0 μg·m L-1 for gallic acid,4.0~40 μg·m L-1 for protocatechuic acid and 2.0~20μg·m L-1 for catechin. The correlation coefficients were between 0.9998~ 0.9999. The average recoveries(n=9) of three phenolic acids components were 104.70%,99.54%,105.84%.The developed method is simple, rapid and efficient, which is suitable for the determination of three phenolic acids components in pine needles of C. deodara, but also will provide an important basis for the follow–up research.2.The content determination and study on the total polyphenol ultrasonic extraction from pine needles of Cedrus deodara.Gallic acid is used as standard substance,using ferrous tartrate spectrophotometric determination of total polyphenol from pine needles of Cedrus deodara at a wavelength at 550 nm.Response surface methodology(RSM) was used to optimize the ultrasonic-assisted extraction process of total polyphenol from pine needles of Cedrus deodara based on single-factor experiment and the content of total polyphenol as indicators.The three factors including ethanol concentration,extraction time,material-solvent ratio.The results show that gallic acid had good liner relationship at 0~1265μg(r=0.9997).Theaverage recovery and RSD was 103.10% and 1.99%.The optimal condition was as follow:the extraction solvent was 55% ethanol, extraction time was 65 min,material-solvent ratio was1:20(g/m L),ultrasonic extraction times was twice.Under the above conditions,the extraction rate of total polyphenol reached 5.46%.3.Study on purification of total polyphenol from Pine Needles of Cedrus deodara by macroporous resin.Using the dynamic adsorption rate of total flavonoids from pine needles of Cedrus deodara as an index screening of macroporous resin,and evaluation of the screening resin separation of total polyphenols process.The total polyphenols was analyzed by ultraviolet spectrophotometry. HPD-600 was the most suitable type of macroporous resin and the optimal separation conditions were as follows:sample concentration was 1.5 mg/m L(8 BV)with a speed of 2 BV/h, the PH range was 4.0 to 5.0,elunt concentration was 70% alcohol(3BV) with a speed of 4 BV/h.Under the above conditions,the total polyphenol powder purity from 9.88% to 34.65%.4.Total polyphenols from Pine Needles of Cedrus deodara Antioxidant activity evaluation.VC and BHT as standard substance,DPPH and ABTS free radical-scavenging assay were used to test the antioxidant activity of total polyphenol extractive from Pine Needles of Cedrus deodara.Using the C. elegans model, through determination the ROS level of C. Elegans to evaluate the antioxidant capacity of total polyphenols extractive from Pine Needles of Cedrus deodara.The results show that the total polyphenols had strong scavenging effect on DPPH free radicals, ABTS radical,the ability of purified total polyphenols extract removing DPPH free radicals is similar with VC;total polyphenol significantly reduced the level of reactive oxygen species of C. elegans, shows that its antioxidant activity. |
PDF Full Text Request