摘要
本试验研究了万寿菊叶黄素的分离方法及分离条件。采用高效液相色谱法（HPLC）测定叶黄素的含量,色谱柱为岛津VP-ODSC18柱（150mm×4.6mm,5μm）,检测波长454nm;以甲醇-0.1%磷酸溶液为流动相进行梯度洗脱,流速1.0mL/m in;柱温30℃;进样量10μL;校准曲线法定量,在0.6-2.0μg范围内具有良好的线性关系,回归方程为Y=1.16×10^7C＋4.99×10^4,相关系数r=0.9984。采用柱层析法分离万寿菊叶黄素的皂化液,实验结果表明：固定相采用硅胶G（100-200目）;胡萝卜素洗脱剂为V（石油醚）∶V（乙酸乙酯）=5∶1;叶黄素洗脱剂为V（石油醚）∶V（乙酸乙酯）=1∶1;层析柱规格为Φ26×510mm;使用皂化液直接湿法上样;吸附时间为30m in;上样量为0.4mL皂化液/g硅胶;柱温为常温;皂化液可直接上样;在以上条件下,经分离回收溶剂后得到含量为83.3%的叶黄素产品。该方法有良好的分离能力,便于操作,产品纯度稳定。硅胶在重复使用5次以内,对分离效果的影响不明显。
This paper studied the method and conditions of separating lutein from Marigold.High performance liquid chromatography（HPLC）was applied to determine lutein content.The operating parameters were：Shimadzu VP-ODSC18column（150mm×4.6mm,5μm）,detection wavelength454nm;methanol-0.1%phosphoric acid as mobile phase gradient elution,flow rate1.0mL/min;column temperature30℃;injection volume 10μL.Chromatogram peak area（Y）against the injection mass concentrations（C）of lutein was in good linearity in the range of0.6-2.0μg,regression equation was Y=1.16×10^7C＋4.99×10^4,the correlation coefficient r=0.9984.The results showed that：the stationary phase was silica gel G（100-200mesh）;carotene eluting agent was V（petroleum ether） ∶V（ethyl acetate）=5 ∶1;lutein eluting agent was V（petroleum ether） ∶V（ethyl acetate）=1 ∶1;chromatography column was Φ26×510mm;saponification liquid was used directly on the wet sample;adsorption time was 30min;sample volume was 0.4mL saponification liquid/g silica gel;column temperature was room temperature;saponification solution added directly on the sample;in the above conditions,the separation and recovery of the lutein was 83.3%.This method separated optimally,was easy to operate and the products were stable.Silica gel was reused 5 times and showed no obvious differences on the separation effect.
出处
《中国食品添加剂》
CAS
北大核心
2010年第3期51-55,130,共6页
China Food Additives
关键词
叶黄素
万寿菊
分离
柱层析
lutein
separation
column chromatograph
marigold.
作者简介
张志强（1969-），男（汉），副教授，本科，从事天然产物提取与分离研究。
