芥菜疙瘩红色素的制备及其稳定性与抗氧化活性

Preparation,Stability,and Antioxidant Activity of Red Pigment from Brassica napiformis

通过单因素试验对芥菜疙瘩红色素的制备条件进行优化,使用AB-8大孔吸附树脂对其进行纯化;分别研究pH值、光照以及食品添加剂对红色素稳定性的影响;通过检测该色素对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸[2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS]阳离子自由基和羟自由基的清除能力以及对中波紫外线(ultraviolet radiation b,UVB)照射损伤的人表皮角质形成细胞(HaCaT)的保护作用,来探究其抗氧化活性。结果表明:芥菜疙瘩红色素制备的最佳温度为80℃,最适加热时间为3 h,乙醇提取的最适浓度为80%(体积分数),通过AB-8大孔吸附树脂纯化后,获得芥菜疙瘩红色素,其得率为(3.26±0.014)%。芥菜疙瘩红色素在酸性环境中保持稳定,微酸近中性时色素保存率显著降低;高温光照显著降低红色素溶液的稳定性,但对固体状态的红色素影响较小;糖类添加剂和抗氧化剂对红色素无明显影响,Mg^(2+)、Ca^(2+)、K^(+)以及柠檬酸具有增色作用,而焦亚硫酸钠会显著降低色素溶液保存率。该色素对DPPH、ABTS^(+)、羟自由基均具有显著的清除能力;通过提高UVB损伤细胞(HaCaT)中超氧化物歧化酶(superoxide dismutase,SOD)活性和还原型谷胱甘肽(glutathione,GSH)含量,降低丙二醛(malondialdehyde,MDA)含量的方式发挥抗氧化和保护细胞的作用。

The preparation conditions of red pigment from Brassica napiformis were optimized by single factor experiment and the resultant red pigment was then purified by AB-8 macroporous adsorption resin.The effects of pH,light,and food additives on the stability of red pigment were studied.The scavenging activity of B.napiformis red pigment against 1,1-diphenyl-2-picrylhydrazyl(DPPH)radicals,2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)cation radicals,and hydroxyl radicals,and its protective effects on human HaCaT keratinocytes damaged by medium-wave ultraviolet ray(UVB)were detected to explore its antioxidant activity.The results showed that the optimal temperature was 80℃,the optimal heating time was 3 h,and the optimal concentration of ethanol extraction was 80%(volume fraction).After purification by AB-8 macroporous adsorption resin,the B.napiformis red pigment was obtained with a yield of(3.26±0.014)%.The red pigment remained stable in the acidic environment,and the preservation rate decreased significantly in the slightly acidic environment close to neutral.Illumination at a high temperature significantly reduced the stability of red pigment solution,but slightly affected the red pigment in solid state.Sugar additives and antioxidants had no obvious effect on red pigment.Mg^(2+),Ca^(2+),K^(+),and citric acid had a hyperchromic effect on the pigment,while sodium metabisulfite significantly reduced the preservation rate of the pigment solution.The red pigment showed significant scavenging abilities against DPPH,ABTS^(+),and hydroxyl free radicals.It exerted an antioxidant and protective of cell effect by potentiating the activity of superoxide dismutase(SOD)and the content of reduced glutathione(GSH)in UVB-damaged HaCaT cells and reducing the content of malondialdehyde(MDA).