| Tea seed oil is a mature seed of Camellia Camellia,a vegetable seed obtained from tea seeds.Tea seed oil is rich in unsaturated fatty acids and can be stored for 1-2 years at room temperature.As a new edible oil in China,tea seed oil has been widely concerned at home and abroad.However,there are few reports on the separation and purification of polar concomitants in tea seed oil and the antioxidant activity of their monomeric substances.It is impossible to explain the structure ofpolar concomitants in tea seed oil and its role in the oxidation of oils.Therefore,this paper takes tea seed oil as the research object.Firstly,the effects of different extraction methods on the polar concomitant of tea seed oil and the extraction process of polar companion of tea seed oil are optimized.Secondly,the macroporous resin AB-8 is used to enrich the polar concomitant,and the polar concomitant monomer is separated and purified by column chromatography combined with preparative liquid chromatography.Finally,the antioxidant activity of polar concomitant substances is studied by in vitro chemical exqeriments and cell models,and the monomeric substances with high antioxidant activity are screened.Obtain the following results:(1)Taking tea seed of Echa No.10 as the research object,comparison of the effects of solvent,alkali,acid and enzymatic methods on the polar concomitant of extracted tea seed oil.After alkaline hydrolysis,the polar composition of tea seed oil is extracted and the antioxidant activity is the highest,the EC50 value is 125.37±1.28μM.The optimal extraction process for the polar companion of tea seed oil is determined by alkaline hydrolysis method.The concentration of NaOH was 2.0 mol/L,the ratio of material to liquid is 1:5,the extraction temperature is 45℃,and the extraction time is 3 h.The pH is adjusted to 2 after alkaline hydrolysis.The phenolic mass of the tea seed oil polar concomitant extracted under this condition is 118.35±1.65 mg/kg,and the EC50 value is 112.83±1.34 μM.(2)Enrichment and preliminary separation of the polar inclusions of tea seed oil with macroporous resin AB-8.The optimal separation and purification conditions are as follows:the sample concentration is 0.5 mg/mL,the loading volume is 4 BV,the loading flow rate is 1 BV/h,the eluent ethanol volume fraction is 60%,and the elution flow rate is 1 BV/h,eluent volume is 5 BV.Under the condition of this process,the purity of the tea seed oil polar concomitant is increased from 45.83%to 68.53%,and the recovery rate of the polar concomitant is 78.83%.Macroporous resin is eluted in sections to obtain four components.The optimum conditions for the preparation of component 1 by preparative liquid chromatography are:the injection volume is 60 mL and the elution flow rate is 20 mL/min.;the optimum process conditions for the separation of components 2 and 3 are as follows:injection volume is 80 mL and elution flow rate is 15 mL/min.Finally,6 kinds of compounds 1,3,5,10,13,18 are obtained by column chromatography on Sephadex LH-20 Sephadex.After liquid chromatography analysis,it was preliminarily concluded that compound 1 was ferulic acid and compound 13 was cinnamte caffeate.(3)The antioxidant activities of the polar companion monomers of tea seed oil are studied by chemical method(DPPH,ABTS method)and cell antioxidant(CAA)exqeriments.All of the above methods show that the six polar concomitant monomers have certain antioxidant properties.The CAA antioxidant capacity values of compounds 1,3,5,10,13,and 18 were 42.89±2.34,8.46±0.23,12.08±0.29,20.45±1.89,50.22±2.75,and 10.33±0.9μmol quercetin equivalent/100,respectively.Antioxidant ability:Compound 3>Compound 5>Compound 1>Compound 18>Compound 13>Compound 10(p<0.05). |
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