Anesi Andrea, Guella Graziano
Bioorganic Chemistry Lab, Department of Physics, University of Trento, Via Sommarive 14, 38123 Povo (Trento), Italy.
Bioorganic Chemistry Lab, Department of Physics, University of Trento, Via Sommarive 14, 38123 Povo (Trento), Italy; Biophysical Institute, CNR, Via alla Cascata 56/C, 38123 Povo (Trento), Italy.
J Chromatogr A. 2015 Mar 6;1384:44-52. doi: 10.1016/j.chroma.2015.01.035. Epub 2015 Jan 21.
In this paper, we report the development of a new method based on HILIC-ESI-MS for the separation of several different membrane lipid classes and their detection on a triple quadrupole mass spectrometer using Precursor Ion (PIS) and Neutral Loss (NL) scanning in positive ion mode. Four different columns were tested for their ability to separate, under different conditions, a mixture of 14 lipid standards containing 7 glycerophospholipids (GPL), 2 glycosphyngolipids (GSL), 3 glycolipids (GL) and 2 betaine lipids (BL). The best separation was obtained using a Lichrosphere DIOL column as stationary phase and water (10mm ammonium acetate)/acetonitrile gradient elution as mobile phase which allows the separation of the 14 lipid classes within 35min runtime. Our method was successfully tested for the separation and analysis of crude lipid extracts obtained from a green alga (Jaoa bullata), a dinoflagellate (Peridinium cinctum) and a plant (Vitis vinifera cv. Corvina).
在本文中,我们报告了一种基于亲水相互作用液相色谱-电喷雾电离质谱(HILIC-ESI-MS)的新方法的开发,该方法用于分离几种不同的膜脂类别,并在正离子模式下使用前体离子(PIS)和中性丢失(NL)扫描在三重四极杆质谱仪上进行检测。测试了四种不同的色谱柱在不同条件下分离14种脂质标准品混合物的能力,该混合物包含7种甘油磷脂(GPL)、2种糖鞘脂(GSL)、3种糖脂(GL)和2种甜菜碱脂(BL)。使用Lichrosphere DIOL色谱柱作为固定相,水(10mM醋酸铵)/乙腈梯度洗脱作为流动相可实现最佳分离,该条件能在35分钟的运行时间内分离出14种脂质类别。我们的方法成功用于从绿藻(Jaoa bullata)、甲藻(Peridinium cinctum)和植物(酿酒葡萄品种科维纳)中获得的粗脂质提取物的分离和分析。