葡聚糖凝胶Sephadex LH-20柱层析分离纯化蓝莓花色苷的研究
详细信息Study on isolation and purification optimization of blueberry anthocyanins by Sephadex LH-20
-
摘要: 本论文初步探讨了采用Sephadex LH-20凝胶柱层析分离纯化蓝莓花色苷的工艺条件,比较了花色苷在不同缓冲液浓度、不同流速和不同上样量条件下的分离效果,并将最佳分离条件下得到的组分进行紫外可见波长扫描,对其中的花色苷组分进行HPLC检测。结果表明,当以30%的酸化甲醇为洗脱液,采用混合流速进行洗脱,进样量为20 mg时,蓝莓花色苷的分离效果最好,共收集到七个峰组分。经紫外-可见光谱扫描,可确定组分Ⅰ、Ⅱ、Ⅲ为非花色苷类物质;组分Ⅳ、Ⅴ、Ⅵ、Ⅶ为花色苷类物质。由HPLC检测可知:组分Ⅳ中只含一种蓝莓花色苷;组分Ⅵ、Ⅶ均以一种花色苷为主,且其峰面积比例达到85%以上;组分Ⅴ中主要含两种花色苷。
-
关键词:
- 蓝莓花色苷 /
- Sephadex LH-20 /
- HPLC
Abstract: The extraction and purification of blueberry anthocyanins by Sephadex LH- 20 were preliminarily disscussed.The respective separating effects of different buffer concentration,flow velocity and sample loading volume were compared and the optimization of the purification condition were selected. The sample components which separated by the optimal condition were scanned on UV- Visible wavelength and identified by HPLC analysis.The results showed that optimal extraction of blueberry anthocyanins conditions were as following : eluant was 30% ethanol,eluting method was mixture velocity elution,sample loading volume was 20 mg,and seven segments can be collected. Based on the scanning of UV- Visible wavelength,segment Ⅰ,Ⅱ,Ⅲ were not anthocyains and segment Ⅳ,Ⅴ,Ⅵ,Ⅶ were anthocyains.HPLC analysis results showed that segment Ⅳcontained one anthocyanin,segment Ⅵ,Ⅶ mainly contained one anthocyanin respectively and the peak area ratio reached over 85%,segment Ⅴ mainly contained two anthocyanins.-
Keywords:
- blueberry anthocyanins /
- Sephadex LH-20 /
- HPLC
-
[1] 解利利,张慜,孙金才.化学吸附法脱除蓝莓汁中单宁的研究[J].食品与生物技术学报,2010,6:847-853. [2] 马艳萍.蓝莓的生物学特性、栽培技术与营养保健功能[J].中国水土保持,2006,2:47-49. [3] 李咏梅.越橘的开发利用价值[J].特种经济动植物,2002,6:23. [4] 杨秀松.蓝莓汁生产热烫工艺对多酚氧化酶及多酚类化合物的影响[J].化学工程师,2012,5:54-59. [5] 吕春茂,王新现,包静,等.越橘果实花色苷的体外抗氧化性[J].食品科学,2010,23:27-31. [6] Takanori T,Mie W,Katsumi O,et al.Antioxidative Activity of the Anthocyanin Pigments Cyanidin 3-O-.beta.-D-Glucoside and Cyanidin[J].Journal of Agricultural and Food Chemistry,1994,42:2407-2410.
[7] Taverniti V,Fracassetti D,Del Bo C,et al.Immunomodulatory Effect of a Wild Blueberry Anthocyanin-Rich Extract in Human Caco-2 Intestinal Cells[J].Journal of Agricultural and Food Chemistry,2014,62(33):8346-8351.
[8] Yun J W,Lee W S,Kim M J,et al.Characterization of a profile of the anthocyanins isolated from Vitis coignetiae Pulliat and their anti-invasive activity on HT-29 human colon cancer cells[J].Food and Chemical Toxicology,2010,48(3):903-909.
[9] Cai H,Marczylo T H,Teller N,et al.Anthocyanin-rich red grape extract impedes adenoma development in the Apc Min mouse:Pharmacodynamic changes and anthocyanin levels in the murine biophase[J].European Journal of Cancer,2010,46(4):811-817.
[10] Cassidy A,Kenneth J M,Lydia L,et al.High Anthocyanin Intake Is Associated With a Reduced Risk of Myocardial Infarction in Young and Middle-Aged Women[J].Circulation,2013,127(2):188-196.
[11] Roopchand D E,Kuhn P,Rojo L E,et al.Blueberry polyphenol-enriched soybean flour reduces hyperglycemia,body weight gain and serum cholesterol in mice[J].Pharmacological Research,2013,68(1):59-67.
[12] 陈介甫,李亚东,徐哲.蓝莓的主要化学成分及生物活性[J].药学学报,2010,4:422-429. [13] Seiji Me,Noriko T,Mariko S,et al.Vision preservation during retinal inflammation by anthocyanin-rich bilberry extract:cellular and molecular mechanism[J].Laboratory Investigation,2012,92(1):102-109.
[14] Canter P H,Enst E.Anthocyanosides of Vaccinium myrtillus(bilberry)for night vision:A systematic review of placebocontrolled trials[J].Survey of Ophthalmology,2004,49(1):38-50.
[15] 吕春茂,包静,孟宪军,等.HPD-700型大孔树脂对野生越橘花色苷分离的研究[J].食品科学,2012,10:78-83 [16] 郑红岩,于华忠,刘建兰,等.大孔吸附树脂对蓝莓花色苷的分离工艺[J].林产化学与工业,2014,4:59-65. [17] 李颖畅,郑凤娥,孟宪军.大孔树脂纯化蓝莓果中花色苷的研究[J].食品与生物技术学报,2009,4:496-500. [18] 刘玮,钱慧碧,辛秀兰,等.蓝莓果渣中总黄酮的提取纯化及抗氧化性能的研究[J].食品科技,2011,2:216-219. [19] Wang E,Yin Y,Xu C,et al.Isolation of high-purity anthocyanin mixtures and monomers from blueberries using combined chromatographic techniques[J].Journal of Chromatography A,2014,1327(31):39-48.
[20] 朱振宝,吴园芳,易建华.紫甘蓝花色苷的分离纯化[J].食品科技,2012,6:239-243. [21] 田瑶.蓝莓中黄酮类物质的提取、分离纯化及生物活性研究[D].东北农业大学,2012. [22] 孙建霞,张燕,孙志健,等.花色苷的资源分布以及定性定量分析方法研究进展[J].食品科学,2009,5:263-268.
计量
- 文章访问数:
- HTML全文浏览量:
- PDF下载量:
下载:
