高效液相色谱法测定栅藻中5种类胡萝卜素的含量

Determination of Five Carotenes in Scendesmus by High Performance Liquid Chromatography

建立了栅藻中虾青素、叶黄素、玉米黄质、角黄素和β-胡萝卜素5种类胡萝卜素的C30-反相高效液相色谱测定方法。冻干的藻粉经二甲基亚砜、二氯甲烷∶甲醇（25∶75,V/V）混合溶剂超声提取后,经0.107mol/L NaOH-甲醇溶液4℃下黑暗冷皂化12h,采用YMC-carotenoid C30色谱柱（250×4.6mm,5μm）,以甲醇、甲基叔丁基醚、水组成的流动相梯度洗脱,紫外检测器450nm检测。在优化实验条件下,5种色素可以很好地从藻粉中提取出来并且转化为游离的类胡萝卜素,并且在不同浓度下其峰强度和质量浓度呈良好的线性。虾青素、叶黄素、玉米黄质、角黄素和β-胡萝卜素的平均回收率范围为101%-107%,平均相对标准偏差范围为0.49%-2.78%,检出限范围为0.05-0.3μg/mL。该方法操作简单准确、可靠,可同时定量分析栅藻中上述5种类胡萝卜素的含量。

A C30-high performance liquid chromatographic（C30-HPLC）method was used for the determination of carotenes in scendesmus.Freeze-dried algae powder sample was ultrasonically extracted with dimethyl sulfoxide,dichloromethane-methanol（25∶75,V/V）,and saponified with 0.107mol/L NaOH-methanol solution at 4℃in dark for 12 hours.The separation of carotenes was achieved using a YMC-Carotenoid C30（250×4.6mm,5μm）column combined with a mobile phase consisting of methanol,tert-Butyl methyl ether and water.The UV detection was operated at 450 nm.In this study,five pigments from algae powder were coverted to free carotenes completely,resulting in a good linear relationship between chromatographic peak areas and the concentrations.The average recovery of astaxanthin,lutein,zeaxanthin,canthaxanthin andβ-caortene was between 101% and107%.The relative standard deviations of the method were between 0.49% and 2.78%,while the detection limits were in the range of 0.05-0.3μg/mL.The results showed that the method is accurate,reliable with simple operation for the determination of five carotenoids in scenedesmus.