Study On Compound Extraction Process Selenium-containing Compounds From Cardamine Hupingshanensis And In Vivo Anti-oxiolation Activity Of Selenium Containing Flavone

In this paper,the extraction technology of selenium-containing organic compounds(proteins,polysaccharides and flavonoids)from hyperselenium plants(Cardamine hupingshanensis)was studied.In order to make full use of selenium in Cardamine hupingshanensis,enzymatic hydrolysis of the extracted residue was carried out and the optimum process was obtained.At the same time,Cardamine hupingshanensis was cultured with exogenous selenium and its flavonoids were extracted,and its functional properties were studied.The main conclusions are as follows:1.Protein,polysaccharide,flavonoids and selenium were determined by phenol sulfate method,Coomassie brilliant blue colorimetry,aluminum salt colorimetry and atomic fluorescence spectrometry.A set of comprehensive utilization technology of Cardamine hupingshanensis was optimized by single factor and orthogonal design.The extraction process is as follows:(1)The extraction process of low-temperature protein was as follows: extraction temperature 50 ?,extraction time 3 h,material-liquid ratio 1:25 g/mL,extraction times 1 time.(2)The high-temperature water extraction technology of polysaccharide was as follows: the ratio of material to liquid was 1:35 g/mL,the extraction temperature was 90 ?,the extraction time was 3h,and the extraction times were once.(3)High temperature ethanol extraction of flavonoids process for 4 hours,extraction temperature 85 ?,material-liquid ratio1:30 g/mL,ethanol concentration 50%.The results showed that the protein content of Chinese Cardamine hupingshanensis was 23.96 mg/g,the selenium content of protein was 125.77 ?g/g,the polysaccharide content was 36.88 mg/g,the selenium content of polysaccharide was 29.81 ?g/g,the flavonoid content was 7.57 mg/g,and the selenium content of flavonoid was 9.26?g/g2.The enzymatic hydrolysis of Cardamine hupingshanensis residue with ellulose and pectinase was studied,and the optimum process was optimized by single factor and orthogonal test.The results showed that: the optimum enzymatic hydrolysis conditions of ellulose were as follows: enzymatic hydrolysis temperature60 ?,cellulose concentration 50 U / mL,enzymatic hydrolysis pH = 5.0,enzymatic hydrolysis time 90 min.Under these conditions,the dissolution rate of selenium was51.32%.The optimum enzymatic hydrolysis process of pectinase was as follows:enzymatic hydrolysis time 90 min,enzymatic hydrolysis temperature 40 ?,enzymatic hydrolysis pH=3.5,pectinase concentration 45 %.Under these conditions,the dissolution rate of selenium was 30.21%.3.The antioxidant capacity of Flavonoids from Cardamine hupingshanensis was determined by the free radical scavenging effect in vitro.Cardamine hupingshanensis flavonoids containing selenium had a certain scavenging effect on free radicals.Except for superoxide anion,the other free radicals showed good antioxidant effect and showed a good linearity with the increase of dosage.The scavenging effect on hydroxyl radical was significantly higher than that of rutin and slightly higher than that of Vc.At low concentration,DPPH scavenging effect was significantly higher than that of rutin and slightly lower than that of Vc,and at high concentration,DPPH scavenging effect was almost the same as that of Vc.In the low concentration range,the total reducing power of common flavonoids and Vc was not significantly different,and was significantly lower than that of selenium-containing flavonoids;in the high concentration,the total antioxidant capacity of the two crude flavonoids and Vc were basically the same.Using Vc and rutin as positive control,SOD,CAT,GSH-Px activity and MDA content as determination index,mice were tested.Results show:Both flavonoids of Cardamine hupingshanensis can increase SOD,CAT,GSH-Px activity and decrease MDA content in mice with good linear effect.The antioxidant effect of crude flavonoids containing selenium was significantly higher than that of crude flavonoids,similar to that of positive control Vc.When the dosage reached 100 mg/kg,the mice in the rutin test group and crude flavonoid test group lost weight and showed slight toxicity,while the mice in the selenium-containing crude flavonoid treatment group had no significant difference in body weight,indicating that selenium had the effect of enhancing immunity.4.Different concentrations of selenium-containing crude flavonoids fromCardamine hupingshanensis and common crude flavonoids from Cardamine hupingshanensis were used to treat murine melanoma cell line B16 and human lung cancer cell line A549 respectively.Reaction time has a certain effect on the proliferation of cancer cells,the overall test in 12 hours to 24 hours with the increase of time,the inhibition rate of cancer cells significantly increased;when 24 hours after the inhibition rate of some experimental groups has no significant effect,or even a downward trend,indicating that the inhibition effect on cancer cells has reached the maximum at this time.To a large extent,with the increase of time,the proliferation rate of cancer cells gradually increased;compared with flavonoids containing selenium and common flavonoids,selenium can increase the inhibitory effect of flavonoids on cancer cells,and prolong the action time.