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Studies On Preparation Of Proanthocyanidin From Pine Needles And Their Biological Activities

Posted on:2013-12-20Degree:MasterType:Thesis
Country:ChinaCandidate:Q L HuangFull Text:PDF
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Pine needles is one of the main byproduct of the Pinaceae. Pine needles as the speed of a renewable, can be harvested throughout the year, are widely distributed, the natural accumulation of large, sustainable use of natural renewable resources, the volatile components do research at home and abroad, but The water-soluble composition of pine needles is not enought, so the water-soluble composition of pine needles is not enough. The pine is a rich plant resources of Zhejiang Province, the distribution area of many types of masson pine, slash pine and black pine. The study use pine resources of Zhejiang Province to research on the pine needles separation of procyanidins preparation process as well as its biological activity.The main achievement of this article are as follows:1. By literature review, internet search, field surveys and visits, etc, to master the distribution of resources of the province to pine, and then conduct field sampling to determine the three pines resources:Pinus elliottii Engelm,Pinus massoniana Lamb and Pinus thunbergii Parl. Final the pine needles of Pinus elliottii Engelm as experimental materials, a comparative study of the sulfuric acid-vanillin and butanol-hydrochloric acid method to determine the sulfuric acid-vanillin colorimetric conditions as follows:30%of concentrated sulfuric acid,4%of the vanillin reaction temperature30℃, reaction time15min; butanol-hydrochloric acid method of colorimetric conditions:100℃reaction temperature, reaction time45min. However, considering the sulfuric acid-vanillin method with good reproducibility, and is simple, took less advantage of the subject, procyanidins determination of sulfuric acid-vanillin method.2. The use of sulfuric acid-vanillin method for the determination of the Pinus massoniana Lamb,Pinus elliottii Engelm and Pinus thunbergii Parl. three kinds of extract proanthocyanidins content, determine the measured wet to loose needles for optimum extraction of raw materials. Then the effect of ultrasonic assisted extraction, solvent extraction methods such as extraction of wet pine needles proanthocyanidins, and finally through the design of a single factor and orthogonal experiments to determine the best conditions of the extraction process:Water extraction of pine needles proanthocyanidins, extraction temperature of100℃, extraction time was150min, the pH value of4, solid to liquid ratio of1:14(g:ml), measured under the conditions of this process,the proanthocyanidins yield was (3.3415±.0286)%. Finally, pine needles proanthocyanidins crude was purified with AB-8macroporous adsorption resin Research AB-8big hole adsorption resin procyanidins adsorption, parsing attached characteristics, determination of the content of procyanidins by sulfuric acid-vanilla aldehyde method filter get the best purification process:water elution rate of0.8ml/min,1.5mL of sample volume, elution20%of the alcohol concentration, alcohol elution flow rate of1.20ml/min.3. Through the determination of T-AOC and of OH·,·O2-,DPPH·free radicals such as anti-oxidation indicators, and combined with VC and VE in control, studies have shown that pine needles of the proanthocyanidins on the DPPH·the free radical scavenging capacity was significantly higher than VC and vitamin E, and17.347μg/mL to reach the ICSO.Pine needles proanthocyanidins, total antioxidant capacity and the hydroxyl radical, superoxide anion scavenging capacity of lower than VC, but the pine needles proanthocyanidins antioxidant stability was significantly higher than VC.The total antioxidant capacity of VC on pine needles proanthocyanidins have a synergistic effect is used alone16times the proanthocyanidins,7.8times VC alone.4. By observing the proanthocyanidins of different doses of pine needles in mice blood, liver, brain and heart tissue of the T-AOC, MDA, GSH-PX and SOD levels and to explore its mouse tissueantioxidant effects on cell viability, studies have shown that pine needles proanthocyanidins can effectively enhance the mouse tissue SOD, GSH-PX activity, lower T-AOC, MDA content, but also helps the mice to maintain a strongantioxidant capacity, and promote cell proliferation activity in a certain concentration.
Keywords/Search Tags:pine needles proanthocyanidins, separation, purification, antioxidant, MTT
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