葵花籽壳黑色素提取鉴定及抗氧化性研究
Study on extraction, identification and antioxidant activity of melanin of sunflower seed shell
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摘要: 利用碱提酸沉和超声波等方法从葵花籽壳中提取黑色素,Folin-Ciocalteu法测定葵花籽壳黑色素粗提取物中总酚的含量,采用单因素和正交实验确定葵花籽壳黑色素提取的最佳工艺条件,紫外和红外光谱法初步鉴定黑色素;以VC、没食子酸为对照,测定黑色素粗提物与纯化黑色素的还原力和清除DPPH.的能力。结果表明,碱提酸沉葵花籽壳黑色素粗提物呈黑色、无光泽,最优提取条件下氢氧化钠浓度为0.5mol/L,提取时间1.5h,料液比1:25,提取温度25℃下,黑色素粗提物的得率为12.37%(RSD2.14%);纯化的黑色素呈黑色、有金属光泽,得率为2.07%(RSD9.27%);粗提物总酚含量以没食子酸计为0.48%。葵花籽壳黑色素亚硫酸钠溶液在292nm波长有最大吸收峰,且吸光度与黑色素的浓度呈正相关,红外光谱图有黑色素红外吸收特征。黑色素的还原力和清除DPPH.自由基的能力明显比VC低,在同一浓度下,纯化前后黑色素的抗氧化性能力无明显改变。Abstract: The melanin of sunflower seed shell (MS) was extracted based on its characteristics of dissolving in alkaline solution and precipitating in acid solution assisted with ultrasonic.Folin-Ciocalteu (FC) method was used to determine total phenol content, single factor and orthogonal tests were used to determine the optimum extract conditions of MS, UV and infrared spectroscopies were used to identify the structure, the reducing power and ability of scavenging DPPH·were used to evaluate the antioxidant activity against the same concentration of VC and gallic acid.The results showed that optimum extract conditions were 0.5mol/L NaOH, time for 1.5h., solid-liquid ratio of 1∶25 at 25℃, the crude extract of MS extracted by alkaline and acid was black and lacklustre with a yield of 12.37%, RSD 2.14%, and the purified MS was metallic luster with a yield of 2.07%, RSD 9.27%.The total polyphenol content of crude extract of MS was 0.48% calculated as GAE.The wavelength of maximum absorption peak was 292nm in the solution of sodium sulfite scanned by Cary 50 UV-Vis spectrophotometer, and absorbance and the concentration of melanin was positively correlated at 292nm.The patterns of classic characteristic absorption of melanin were shown in the ultraviolet and infrared spectrogram, respectively.With the same concentration, the antioxidant capacity of purified melanin was nearly same as that of the crude extract, lower than VC but higer than gallic acid.