玉米蛋白粉中提取叶黄素的工艺研究
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摘要
叶黄素(xanthophylls),是一种类“类胡萝卜素”族物质,包括黄体素(lutein),玉米黄质(zeaxanthin)等。叶黄素是一种性能优异的抗氧化剂,天然的叶黄素对人体具有区别于其它类胡萝卜素的特殊作用,如保护视力、降低白内障发生率等。目前,市场上的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,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。
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 C303μm 4.6×150mm,流动相为甲醇：甲基叔丁基醚(MTBE):水=95：2：3,流速1.0mL/min,柱温30℃,检测波长为450nm/260nm双波长。
建立了以蛋白酶水解预处理、有机溶剂浸提、液液萃取、柱层析、结晶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：采用中性蛋白酶,酶浓度为2.5%,液固比为20mL/g,温度40℃,水解时间5h。获得叶黄素含量为0.3%左右的粗提物。然后用乙醚-粗提液-水(5：5：6)体系进行液液萃取,获得叶黄素收率为98%、含量达到1.0%左右的叶黄素粗制品。在静态吸附实验中,通过考察十种树脂对叶黄素萃取产物的静态平衡吸附量,发现树脂H吸附量相对较大。选用树脂H作为色谱柱固定相进行动态吸附及洗脱实验,以乙醇和水的混合溶剂为流动相进行洗脱,得到叶黄素含量为5.76%、回收率达97.2%的分离产物。将过柱后的产品在氯仿-正己烷溶剂中-4℃下低温结晶,得到纯度64%左右的叶黄素产品
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. Almost all the commercial xanthophylls products are taken from nature. Corn Gluten Meal (CGM) is a by-product from corn starch manufacturing. 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.OmL/min. The wavelength of UV detector was 450nm/260nm and the column temperature was 30℃.
The separation of xanthophylls from CGM consists of three unit operation including extraction, preparative chromatography and crystallization. The proteolytic hydrolysis was explored to enhance the extraction yield of xanthophylls. And then the pigment was extracted by industrial alcohol for getting rid of the insoluble proteins and lipids. After that, ethyl ether was selected as extractant to extract pigment from raffinate with a proportion of ethyl ether:raffinate:water=5:5:6, and the content of xanthophylls in product was increased from 0.3% to 1.0% with a recovery of 98%. Before purification with column chromatography, the performance and separation characteristics relevant to xanthophylls were measured on ten different resins. It was found that macroporous adsorptive resin H offer the largest adsorption abilities among the tested adsorbents and thus was selected as the stationary phase. Using ethanol and water as mobile phase, the content of xanthophylls in product was increased to 5.76%, with a recovery of 97.2%. Finally through crystallization process in the mixture of chloroform and n-hexane at -4℃, the crystalline product was obtained with the purity higher than 64%.
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 C303μm 4.6×150mm,流动相为甲醇：甲基叔丁基醚(MTBE):水=95：2：3,流速1.0mL/min,柱温30℃,检测波长为450nm/260nm双波长。
建立了以蛋白酶水解预处理、有机溶剂浸提、液液萃取、柱层析、结晶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：采用中性蛋白酶,酶浓度为2.5%,液固比为20mL/g,温度40℃,水解时间5h。获得叶黄素含量为0.3%左右的粗提物。然后用乙醚-粗提液-水(5：5：6)体系进行液液萃取,获得叶黄素收率为98%、含量达到1.0%左右的叶黄素粗制品。在静态吸附实验中,通过考察十种树脂对叶黄素萃取产物的静态平衡吸附量,发现树脂H吸附量相对较大。选用树脂H作为色谱柱固定相进行动态吸附及洗脱实验,以乙醇和水的混合溶剂为流动相进行洗脱,得到叶黄素含量为5.76%、回收率达97.2%的分离产物。将过柱后的产品在氯仿-正己烷溶剂中-4℃下低温结晶,得到纯度64%左右的叶黄素产品
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. Almost all the commercial xanthophylls products are taken from nature. Corn Gluten Meal (CGM) is a by-product from corn starch manufacturing. 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.OmL/min. The wavelength of UV detector was 450nm/260nm and the column temperature was 30℃.
The separation of xanthophylls from CGM consists of three unit operation including extraction, preparative chromatography and crystallization. The proteolytic hydrolysis was explored to enhance the extraction yield of xanthophylls. And then the pigment was extracted by industrial alcohol for getting rid of the insoluble proteins and lipids. After that, ethyl ether was selected as extractant to extract pigment from raffinate with a proportion of ethyl ether:raffinate:water=5:5:6, and the content of xanthophylls in product was increased from 0.3% to 1.0% with a recovery of 98%. Before purification with column chromatography, the performance and separation characteristics relevant to xanthophylls were measured on ten different resins. It was found that macroporous adsorptive resin H offer the largest adsorption abilities among the tested adsorbents and thus was selected as the stationary phase. Using ethanol and water as mobile phase, the content of xanthophylls in product was increased to 5.76%, with a recovery of 97.2%. Finally through crystallization process in the mixture of chloroform and n-hexane at -4℃, the crystalline product was obtained with the purity higher than 64%.
引文
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