| Phellinus baumii that also said SangChen or SangEar is a kind of precious medicinal fungi.It mainly distribute in China,Japan,the Philippines,Australia,North America and other places.The main active components of Phellinus baumii are polysaccharide,nonsteroidal,terpenoids and flavonoids,etc.Phellinus baumii has a variety of pharmacological activities,such as antioxidation,anti-tumor,anti-inflamma-tory,hpyerglycemic and anti-microbial,etc.In recent years,antitumor activity has been a hotspot of research.Scholars from all over the world for its antitumor active substances and anti-tumor mechanism are for the in-depth research,promising to develop into a new antitumor drug.At present the study of Phellinus baumii fungi mainly focused on polysaccharide extraction,submerged fermentation condition optimization and the cancer immunology mechanism,etc.And the researches of the flavonoids were rare,but also has a tendency to increase gradually.Based on Phellinus baumii as raw material,the toxicology security experiment were identified in the paper.Ethanol reflow method was adopted to extract Phellinus baumii flavonoids in the paper and through the single factor and response surface method to optimize the extraction process.Phellinus baumii flavonoids are purified by the method of macroporous resin and use LC-MS technology to identify further the Phellinus baumii flavonoids.The antioxidant activity and the protective effect of DNA of Phellinus baumii flavonoids were studied in this paper.The results were as follows:The toxicology tests of Phellinus baumii indicated that the determination result of the acute oral toxicity test in mice belonged to nontoxic range and three genetic toxicity tests were negative.So it preliminarily verified that Phellinus baumii has non-toxic and no genetic toxic effect.Using ethanol extraction of flavonoids in Phellinus baumii by the single factor test and response surface method to get the best process conditions as follows: extracting temperature was 69.95 ?,liquid-solid ratio was 26.10,extraction time was 3.81 h,ethanol volume fraction was 87.16 %.Considering the specific operation,extraction conditions were determined as follows: extraction temperature was 70 ?,liquid-solid ratio was 26,extraction time was 3.8 h,ethanol volume fraction was 87 %.Under this condition,the actual measured flavonoids yield was 14.36 % and was anastomosed with the theoretical prediction that indicated response surface method to optimize the flavonoids in Phellinus baumii was feasible.The crude flavonoids extraction in Phellinus baumii were separated and purified by resin purification.Through static experiments of macroporous resin,the DM301 macroporous resin was chosen best from 6 kinds of macroporous resin to separate and purify flavonoids.The best craft were determined by static and dynamic experiments of macroporous resin for purification of total flavonoids in Phellinus baumii.When the loading quantity of sample was 20 mL,the results were as follows: the sample flow rate was 3 mL/min,the sample volume was 7 BV,eluent was 80% ethanol,elution velocity was 1.5 mL/min,the volume of eluent was 4 BV.The purity of the final 1purification of total flavonoids of Phellinus baumii was 83.27%.By external standard method,we can attain that there were 16.3 mg rutin and 0.13 mg quercetin in 1 g Phellinus baumii flavonoids.The monomer analysis of flavonoids in Phellinus baumii were attained by LC-MS technology,got the 6 kinds of compounds.Through studying on the antioxidant activity of total flavonoids of Phellinus baumii and DNA protection,experimental results showed that total flavonoids of Phellinus baumii can good remove DPPH · free radical,hydroxyl radical and superoxide anion.The restoring force experiment indicated that the reduction power of total flavonoids in Phellinus baumii compared with the same concentration of Vc and BHT were similar.It stated flavonoids in Phellinus baumii has good antioxidant ability.By ultraviolet absorption spectrum of flavonoids interaction with DNA,when flavonoids in Phellinus baumii were joined herring sperm DNA solution,hyperchromic effect was produced.It showed that herring sperm DNA and flavonoids in Phellinus baumii formed complexes.Through the fluorescence spectra,the herring sperm DNA versus flavonoids in Phellinus baumii had obvious quench effect.In the presence of Phellinus baumii flavonoids,bromide in ingot as fluorescence probe,measuring the fluorescence of DNA and EB mixture integral strength,calculated that the constant D was 43.7%. |
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