反相高效液相色谱-电喷雾质谱法测定浒苔中的叶黄素

Determination of Lutein in Enteromorpha prolifera by Reversed-phase High Performance Liquid Chromatography-Diode Array Detector-Electrospray Ionization Mass Spectrometry

建立反相高效液相色谱-电喷雾电离质谱法(RP-HPLC-DAD-ESI-MS)定性、定量测定绿潮藻浒苔中叶黄素的新方法。浒苔样品经超声辅助提取后,采用RP-HPLC-DAD-ESI-MS联用技术对其进行分离鉴定,HPLC采用Eclipse XDB-C18(150mm×4.6mm,5μm)色谱柱,以含0.2%乙酸的甲醇溶液-水为流动相,线性梯度洗脱,检测波长450nm。在选定的最佳仪器条件下,叶黄素与浒苔中的其他化合物分离良好,叶黄素在0.10～20.0mg/L质量浓度范围内具有良好的线性关系(R2=0.9994),能对浒苔提取液中的叶黄素进行准确定量;采用电喷雾质谱联用分析能快速获得叶黄素的分子量信息,可为浒苔中叶黄素的快速鉴别提供依据。

A reversed-phase high performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry（RP-HPLC-DAD-ESI-MS） method was established for qualitative identification and quantitative determination of lutein in Enteromorpha prolifera.Lutein was extracted from Enteromorpha prolifera through ultrasound-assisted extraction,and then analyzed by RP-HPLC-DAD-ESI-MS.HPLC was carried out at the conditions of Eclipse XDB-C18（4.6 mm × 150 mm） column,MeOH-0.2% acetic acid-water（V/V） as the mobile phase with linear gradient elution and detection wavelength of 450 nm.Results indicated that the developed method could be used for the determination of lutein in Enteromorpha prolifera with a good resolution and linear relationship in the range from 0.10 to 20.0 mg/L（R2 = 0.9994）.Molecular weight of lutein was rapidly evaluated by ESI-MS detection.Combined with literature information,lutein in Enteromorpha prolifera could be rapidly identified.This developed method is specific,simple,fast,sensitive and feasible for rapid identification and quantification of lutein,which can eliminate the interference of other pigments and derivatives from Enteromorpha prolifera.