超高效液相色谱法测定万寿菊悬浮培养细胞中的叶黄素  

Determination of Lutein in Cell Suspension Cultures of Tagetes erecta L.by Ultra-performance Liquid Chromatography

作　　者：徐晓辉[1,2] 王莉洁[2] 孙丹[2] 何伟[1,3] 

机构地区：[1]中国中医科学院眼科医院,北京100040 [2]辽宁何氏医学院健康营养学院,沈阳110163 [3]沈阳何氏眼科医院,沈阳110034

出　　处：《天然产物研究与开发》2016年第10期1576-1580,共5页Natural Product Research and Development

摘　　要：建立了超高效液相色谱快速测定万寿菊悬浮培养细胞中叶黄素的方法。样品冷冻干燥后,经甲醇溶液提取,采用Waters ACQUITY UPLCBEH C18色谱柱（50 mm×2.1 mm,1.7μm）分离,柱温为25℃。以乙腈-甲醇（90∶10,v/v）（A）-水（B）为流动相,梯度洗脱,流速为0.4 m L/min。进样体积为5μL,检测波长为448 nm。在1.56-25 mg/L范围内,叶黄素的色谱峰面积与质量浓度之间具有良好的线性关系。采用该方法测定了实际样品中叶黄素的含量,并进行了3个水平的加标回收试验,其回收率为102%-105%,相对标准偏差（n=5）为0.11%-1.19%。该方法快速、操作简单,适合于分析植物培养细胞中叶黄素的含量和产率。A rapid ultra-performance liquid chromatography （UPLC） method was developed for the determination of lutein in cell suspension cultures of Tagetes erecta L.. The samples were prepared by freeze-drying followed by extraction with methanol. The UPLC separation was achieved using a Waters ACQUITY UPLC BEH C18 column （50 mm × 2.1 mm, 1.7 μm） at 25 ℃. The UPLC analysis was performed in the gradient elution mode ,using acetonitrile-methanol （90 ： 10, v/v） （A）-water （B） as mobile phase at a flow rate of 0.4 mL/min. The injection volume was 5 μL and the detection was carried out at 448 nm. A highly significant linear relationship between lutein concentration and peak area was observed over the concentration range from 1.56 to 25 mg/L. The proposed method was applied for the determination of lutein in the actual samples and recoveries were determined at three spiking concentrations. The recoveries ranged from 102% to 105% with relative standard deviations between 0.11% and 1. 19%. The approach was found to be rapid, simple, and was suitable for the analysis of lutein in plant cell cultures to determine the lutein content of the biomass and the lutein productivity.

关 键 词：超高效液相色谱 叶黄素 悬浮培养细胞 万寿菊 

分 类 号：Q813.1[生物学—生物工程] O657.7[理学—分析化学]