| Superfluous free radicals called refuse in organism can have much effect on giant molecules such as protein, lipoid, polysaccharide, nucleic acid. It dam-ages the activity and the structure of normal cell tissue. This will cause many diseases like angiocardiopathy, aging, transformation of normal cells into can-cerous tumor cells, etc. Therefore, getting rid of these superfluous free radicals plays an important role on maintaining body health. Antioxidant may destroy these superfluous free radicals and break the process of chain reaction. The an-tioxidant substances synthesized by chemical method like butylated hy-droxyanisole (BHA), propyl gallate (PG), tertiary butylhydroquinone (TBHQ), etc., which performed excellent activities, have been widely used to food in-dustry, cosmetics and other fields. However, recent-studies showed that synthe-sized antioxidants might have an unfavorable influence on respiratory ferment. Some even cause potential teratogenesis. This is to say, once they get into the body our health will be threatened. Obviously, searching for safe and reliable antioxidants becomes one of the emphatically studying contents for the fine chemicals in the world in recent years. Many plants, especially for the solana-ceae plants resources, contain some antioxidants in various degrees. They can be extracted by scientific way and have safer and more trustful characteristics. It is noticed that extracted antioxidant from the plants are a kind of mixture. Its antioxidant activities are usually weak relative to synthesized antioxidant. And its thermal stability isn't good, especially for the extracts like superoxide dis-mutase (SOD), peroxydase (POD), which is apt to degenerate. These problems seriously hinder the wide application of natural antioxidant. In view of this point, the experiment observed the influences on natural antioxidant activities in the existence of necessary metal elements in the body after systematically studying extraction ways on yields so that obtain dual function antioxidant, which both has antioxidant activities and increases the trace elements, also.The experiments used solanaceae plants such as stalk of S.melongena, ly-cium, and chilli as materials, which have an abundant resource. By experiment, we investigated three extracting ways like extraction, impregnation and ultra-sonic wave on extraction yield of non-enzyme antioxidant and salting out method, precipitation with organic solvents, ultrafiltration fractionation on ex-traction yield of enzyme antioxidant. The optimal extracting method and condi-tion are given. At the same time, the antioxidant activities of enzyme extracts after adding some effectors such as metal ions, Vc, VE and some organic sub-stance were determined and analyzed. The optimal ratios of extracts to effects were chosen. In addition, the thermal stability and storage time for these ex-tracts were also observed. The experimental results were analyzed by general theory of statistics, which showed whether the experimental result had signifi- cant difference and its reliability. The main contents are as following:(1) Extracting methods and types of solvent both affect the effectiveness of anti-oxidant. For chilli, optimal extracting condition is that 85% ethanol is used as extracting solvent, extracting time is 30min by ultrasonic wave. For lyceum, 60% ethanol was selected as solvent, ultrasonic wave irradiating time is 40 min, then, hydrolysis time for the extracts is 3h; For stalk of S.melongena,60% etha-nol is used as solvent and impregnating extraction is selected.The extracting methods also have an influence on activity of superoxide dismutase (SOD) and peroxidase (POD). The activity of SOD obtained through ultrasonic wave extraction, salt out of ammonium sulphate and hyperfiltration is t he highest; the activity of POD obtained through extraction of water, salt out of ammonium sulphate, treatment of acetone and hyperfiltration is the highest. T-test showed that there existed significant difference for these methods. The activity of SOD and POD produced by extracting different materials is not the same. The activity of SOD from chilli is lower than that of lyciun Activity of POD from chilli is stronger than that of lycium.(2) For the same extract, anti-oxidizing property is different due to using different evaluating methods. The non-enzyme extracts from stalk of S.melongena has strong clearing ability for DPPH-, but the ability of anti-oxidization of oil is poorer than BHT. The non-enzyme extracts of chilli has strong abilities in respect of anti-oxidation forβ-renieratene, clearing ability of superoxide anion free radical and anti-oxidation of oil than BHT, but has weak ability for clearing DPPH-. The non-enzyme extracts of lycium has not only strong ability for clearance of DPPH-, but also anti-oxidation for P-renieratene is equivalent to BHT. For SOD the optimal temperature is at 55℃and the optimal pH is 6.5; for POD the temperature is 45℃and pH is 6.0.(3) It was shown by gas chromatography-mass spectrum (GC-MS) that the main composition of the non-enzyme extracts in stalk of S. Melongena are aro-matic series and organic acid compounds such as hexadecanoic acid, phenethylamine, phenylformic acid, dibutyl phthalate, and a small amount of phenolic compounds; for the extracts of chilli, the main compositions are phe-nols and fatty acid compounds like capsaicin,2,4-decadienal,9,12-octadecadi-enoic acid, capsicine, phenylacetic acid,4-hydroxy-3-methylphenol; for the ex-tracts of lycium, the main compositions are such compounds as Oxidation of terpenes, fatty acid ester, aldoketones, aromatic series. There wasn't an obvious dependence between the content of the total phenol in the non-enzyme extracts and yields. However, the suppression ratio of DPPH·and decolorization ratio ofβ-renieratene for non-enzyme extracts seriously depend on the content of the total phenol whose the related coefficient r2 are 0.9927 and 0.9855, respectively. This conclusion is also proved by T-test.(4) The synthetic experiments of extracts and effectors showed that differ-ent types of effectors displayed a great difference. For the non-enzyme extract, trace metal ions can improve the ability of antioxidant, eliminating peroxy radi-cal and DPPH, and oxidation resistance of p-renieratene. The antioxidant of the mixture composed of non-enzyme extracts and vitamin C, which account for 50 wt% in the mixture, is more excellent than synthetics BHT. The antioxidant of non-enzyme extracts for chilli and lycium is greatly enhanced in the existence of selenium ion, which illustrated the existence of synthetic effect. When the concentration of Fe2+ of 800mg/Land Mn2+ of 500mg/L are used and other con-ditions are the same, clearance ability for superoxide radical increases by 55% and 40% for the non-enzyme lycium extracts, and 81% and 79% for the non-enzyme chilli extracts, respectively.Influence of the effectors on activity of POD and SOD isn't the same even if the type of effectors is the same. Such effectors as potassium chloride, sodium chloride, calcium chloride, barium chloride, zinc sulfate, magnesium chloride, zinc acetate, zinc gluconate, manganese gluconate, glycol, propanetriol, mannite and sorbierite are the activating agents for the activity of POD. Such compounds as copper chloride, citric acid are the inhibitors for the activity of POD whose affected tendency conforms to the curve S shape. Binary effectors composed of zinc acetate and sorbierite,which accounts for 50% and its concentration of 2.0 mmol/L is used, increase by 80% activity of POD. Such compounds as glycol, propanetriol and sorbierite are activating agents for the activity of SOD. Such compounds as potassium chloride, sodium chloride, calcium chloride, barium chloride, zinc sulfate, magnesium chloride, zinc acetate, zinc gluconate, man-ganese gluconate are the inhibitors for the activity of SOD. Furthermore, through binary counting backward technique of Michaelis-Menten Equation confirmed that influence of the compounds like zinc gluconate, zinc acetate, calcium chloride, barium chloride, magnesium chloride on SOD belongs to the competing inhibitors and compounds such as potassium chloride, sodium chlo-ride, manganese gluconate, copper chloride and zinc sulfate belongs to anti-competitive inhibitors.(5) The reactive rate equation occurred in the process of oxidizing inactiva-tion for the non-enzyme extracts of chilli and lycium were given out, from which the activation energy of reaction is reduced in the existence of effectors sodium selenite. This can strengthen the ability of getting rid of free radicals of non-enzyme extracts. For non-enzyme extracts of chilli, its rate equation of in-activation is k=1.42×102·e (?), corresponding related coefficient r2 equals 0.99. After effectors sodium selenite was added, rate equation of inactivation is k=0.167·e (?) and related coefficient r2 is 0.958. The activation energy de-creases by 53.8%. For non-enzyme extracts of lycium, inactivation rate kis 1.519·e RT and related coefficient r2 equals 0.908. When effectors was used the inactivation rate k becomes 1.669×10-2·e RT and related coefficient r2 equals 0.981. The activation energy also decreases.For the POD extracted from chilli, its inactivation rate k conforms to 1.36×109·e (?) and related coefficient r2 is 0.96. Once effectors sodium selenite was added into the extracts, inactivation rate k becomes 3.406 x108·e (?) and related coefficient r2 is 0.972. The inactivation energy increases; For SOD, inactivation rate k conforms to 3.612 x103·e (?) and related coefficient r2 is 0.972. After the effectors sodium selenite was added, inactivation rate k is ex-pressed as 2.355×104·e (?). And the related coefficient r2 is 0.904. The activa-tion energy also increases. The activity of POD and SOD was seriously influ-enced by the temperature and easily becomes inactive. The calculation showed that the activity of antioxidant for the chilli extracts at the temperature of 25℃is one tenth of the activity at 4℃because of the loss of activity. For SOD of ly-cium extracts, its activity is one fourth of the activity at the temperature of 4℃due to the loss.(6) The electrical behaviors of POD and SOD were also studied. The re-sults showed that POD and SOD exist with a negative charge in the range of 2 to 9 for pH. The force produced among the particles behaviors strong, and the ag-gregation becomes obvious. The distribution range of the particle size is wider and average diameter is larger; When the effectors were added the distribution range of the particle size is narrow and its average diameter becomes small. The amount of displacement is larger which is beneficial to keeping the extracts sta-ble. And cyclic voltammogram showed that the stability of the electrical polar manufactured by POD-MC/MWCNTs/Pt peak shape is better and peak electrical current is big. |
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