摘要
建立辣椒叶片中类胡萝卜素组分及其含量的分离测定方法.通过对提取溶剂、皂化温度、皂化时间、KOH-乙醇皂化液质量浓度、KOH-乙醇皂化液体积、料液比的选择,以及对流动相、柱温、柱流速等色谱条件的考察,研究提取辣椒叶片类胡萝卜素工艺流程.建立了提取辣椒叶片中类胡萝卜素的适宜工艺和分离测定辣椒叶片中类胡萝卜素组分及其含量的RP-HPLC工艺.技术关键是：以石油醚为提取溶剂、皂化温度为80℃、皂化时间为40min、皂化液质量浓度为20%、皂化液体积为10mL、料液比为1∶5;采用YMC-C30色谱柱（250mm×4.6mm i.d,5μm）;检测波长450nm;柱流速1.0mL·min^-1;柱温25℃;流动相A为乙腈：甲醇（V∶V=60∶40）,流动相B为甲基叔丁基醚（100%）,梯度洗脱.结果表明以外标法定量,叶黄素、玉米黄素和β-胡萝卜素各组分的线性关系良好,相关性系数在0.9961-0.9999之间;平均回收率在85.2%-95.1%之间.该方法为辣椒叶片中叶黄素、玉米黄素和β-胡萝卜素的分离与定量测定提供了方法保证.
A method that can separate and determine the composition and content of carotenoids in pepper leaves is developed.Technological process of extracting catotenoids in pepper leaves is investigated through the researches on the extraction solvent,saponification temperature,saponification time,the concentration and volume of KOH-ethanol saponification solution,ratio of solid-liquid and chromatographic conditions.In this method,the key technology for separation and determination of carotenoids in pepper leaves is the use of the petroleum ether.The saponification temperature is 80 ℃,saponification time is 40 min,the concentration of KOH-ethanol saponification solution is 20%,the volume of KOH-ethanol saponification solution is 10 mL and the ratio of solid-liquid is 1∶5.Separation and determination is achieved by using a YMC C30colum（250mm×4.6mm i.d.,5μm）.A mobile phase of acetonitrile-mthanol（60∶40,v/v）（A）and methyl tert-butyl ether（100%）（B）are eluted with the gradient.Three kinds of carotenoids are resolved with the flow-rate at 1.0 mL·min^-1,column temperature of 25 ℃ and the detection wavelength of 450 nm.The results show that there have good linear correlation for the lutein,zeaxanthin andβ-carotene,and the correlation coefficient is between0.9661 and 0.9999.The average recovery of three carotenoids is between 85.2% and 95.1%.This method can be used widely to separate and determine the components and content of lutein,zeaxanthin andβ-carotene in pepper leaves.
出处
《西北师范大学学报（自然科学版）》
CAS
北大核心
2015年第4期75-80,108,共7页
Journal of Northwest Normal University(Natural Science)
基金
国家自然科学基金资助项目(31260493)
关键词
辣椒叶片
类胡萝卜素
分离测定
方法
pepper leaves
carotenoids
separation and determination
method
