玉米蛋白粉中叶黄素的提取工艺研究
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摘要
叶黄素(xanthophylls)又名“植物黄体素”,属于类胡萝卜素族,包括黄体素(lutein)和玉米黄质(zeaxanthin)等。叶黄素是一种性能优异的抗氧化剂,具有区别于其它类胡萝卜素的特殊生理作用,如保护视力、降低白内障发生率、预防心血管疾病、抗癌以及增强免疫力等。由于叶黄素的化学合成工艺复杂且产品生物活性差,因此,现今叶黄素产品均来自于天然物的提取。玉米蛋白粉(Corn Gluten Meal, CGM)是生产玉米淀粉的副产物,其中的叶黄素含量可达145μg/g。使用玉米蛋白粉作为提取叶黄素的原料,有着资源丰富、副产物再利用的优势,而且能有效提高玉米和玉米蛋白粉的工业附加值,因此相关研究具有重要意义。
本文研究了玉米蛋白粉中叶黄素的提取和分离纯化技术。
建立了叶黄素的反相高效液相色谱分析法,色谱条件为：色谱柱YMC-Carotenoid C30 3um 4.6x150mm,流动相为甲醇：甲基叔丁基醚(MTBE):水=95：2：3,流速1.OmL/min,柱温30℃,检测波长为450nm/260nm双波长。
建立了以表面活性剂预处理、有机溶剂萃取、柱层析等为主的叶黄素提取和分离工艺路线。各过程的最优工艺条件为：采用表面活性剂预处理,表面活性剂浓度为1.0%,液固比为10mL/g,温度为30℃,水解时间为0.5h,获得叶黄素含量为0.3%左右的粗提物；在静态吸附实验中,通过考察HZ816、D630、SD300、S8以及AB8五种树脂对叶黄素萃取产物的静态平衡吸附量,筛选吸附量相对较大SD300树脂作为色谱柱固定相；动态吸附及洗脱实验中,分别以工业酒精和乙酸乙酯作为上样溶剂和洗脱溶剂,最终得到叶黄素含量为6.0%、回收率达80%的分离产物。
Xanthophylls are members of carotenoids family, including lutein and zeaxanthin. As an excellent antioxidants, natural xanthophylls have some properties different from other carotenoids, such as protection of visual acuity, reductuction of incidence of cataract. Since the chemical synthesis process of xanthophylls is complicated and their corresponding biological activity is poor, almost all the commercial xanthophylls products are taken from natural compound. Corn Gluten Meal (CGM) is a by-product from corn starch manufacturing, in which the xanthophylls content is up to 145μg/g. Natural xanthophylls can be obtained from CGM, which has a great annual output.
Xanthophylls were determined by HPLC using YMC-Carotenoid C30 column 4.6mm×150mm (3μm). Mixture of methanol, methyl tertiary butyl ether and water (95:2:3) was used as elution solvent with flow rate of 1.0mL/min. The wavelength of UV detector was 450nm/260nm and the column temperature was 30℃.
The separation of xanthophylls from CGM mainly consists of three operation process including pretreatment, extraction and preparative separation. Comparing with the proteolytic hydrolysis pretreatment method, the surfactant pretreatment of CGM can enhance the extraction ability of xanthophylls by ethanol. Then in the pretreatment process the pigment was extracted by industrial alcohol for getting rid of the insoluble proteins and lipids. Before purification with column chromatography, the performance and separation characteristics relevant to xanthophylls were measured on HZ816、D630、SD300、S8、AB8 resins. It was found that SD300 macroporous adsorptive resins offer the largest adsorption abilities among the tested adsorbents and thus was selected as the stationary phase. Using Ethyl acetate as mobile phase, the content of xanthophylls in product was increased to 6.0%, with a recovery of 80%.
本文研究了玉米蛋白粉中叶黄素的提取和分离纯化技术。
建立了叶黄素的反相高效液相色谱分析法,色谱条件为：色谱柱YMC-Carotenoid C30 3um 4.6x150mm,流动相为甲醇：甲基叔丁基醚(MTBE):水=95：2：3,流速1.OmL/min,柱温30℃,检测波长为450nm/260nm双波长。
建立了以表面活性剂预处理、有机溶剂萃取、柱层析等为主的叶黄素提取和分离工艺路线。各过程的最优工艺条件为：采用表面活性剂预处理,表面活性剂浓度为1.0%,液固比为10mL/g,温度为30℃,水解时间为0.5h,获得叶黄素含量为0.3%左右的粗提物；在静态吸附实验中,通过考察HZ816、D630、SD300、S8以及AB8五种树脂对叶黄素萃取产物的静态平衡吸附量,筛选吸附量相对较大SD300树脂作为色谱柱固定相；动态吸附及洗脱实验中,分别以工业酒精和乙酸乙酯作为上样溶剂和洗脱溶剂,最终得到叶黄素含量为6.0%、回收率达80%的分离产物。
Xanthophylls are members of carotenoids family, including lutein and zeaxanthin. As an excellent antioxidants, natural xanthophylls have some properties different from other carotenoids, such as protection of visual acuity, reductuction of incidence of cataract. Since the chemical synthesis process of xanthophylls is complicated and their corresponding biological activity is poor, almost all the commercial xanthophylls products are taken from natural compound. Corn Gluten Meal (CGM) is a by-product from corn starch manufacturing, in which the xanthophylls content is up to 145μg/g. Natural xanthophylls can be obtained from CGM, which has a great annual output.
Xanthophylls were determined by HPLC using YMC-Carotenoid C30 column 4.6mm×150mm (3μm). Mixture of methanol, methyl tertiary butyl ether and water (95:2:3) was used as elution solvent with flow rate of 1.0mL/min. The wavelength of UV detector was 450nm/260nm and the column temperature was 30℃.
The separation of xanthophylls from CGM mainly consists of three operation process including pretreatment, extraction and preparative separation. Comparing with the proteolytic hydrolysis pretreatment method, the surfactant pretreatment of CGM can enhance the extraction ability of xanthophylls by ethanol. Then in the pretreatment process the pigment was extracted by industrial alcohol for getting rid of the insoluble proteins and lipids. Before purification with column chromatography, the performance and separation characteristics relevant to xanthophylls were measured on HZ816、D630、SD300、S8、AB8 resins. It was found that SD300 macroporous adsorptive resins offer the largest adsorption abilities among the tested adsorbents and thus was selected as the stationary phase. Using Ethyl acetate as mobile phase, the content of xanthophylls in product was increased to 6.0%, with a recovery of 80%.
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