The performance of seven sub-2-μm particle packed columns (2-picolylamine, 2-PIC; charged surface hybrid fluoro-phenyl, CSH-FP; high strength silica C18 SB, HSS-C₁₈; diethylamine, DEA; 1-aminoanthracene, 1-AA; high density diol and ethylene bridged hybrid; BEH) was examined for lipid separation in ultra-high performance supercritical fluid chromatography (UHPSFC) coupled to quadrupole time-of-flight mass spectrometry. Based on the results of the column screening a method for profiling of multiple lipid species from the major lipid classes was developed.

Stationary phases containing β-hydroxy amines, i.e. 1-AA, DEA and 2-PIC, yielded strong retention and poor peak shapes of zwitterionic lipids with primary amine groups, such as phosphatidylserines, phosphatidylethanolamines and its lyso forms. The BEH and HSS-C₁₈ columns showed strong retention of polar and nonpolar lipids, respectively. The Diol column retained the majority of major lipid classes and also produced symmetric peaks. In addition, this column also produced the highest resolution within and between major lipid classes. An injection solvent composed of methanol:chloroform (1:2, v:v) and the addition of 20 mM ammonium formate in the mobile phase improved chromatographic separation and mass spectrometry detection in comparison to ammonium acetate or absence of additive. Finally, chromatographic and mass spectrometric parameters were optimized for the Diol column using a design of experiments approach. The separation mechanism on the Diol column depended on the lipid functionality and the length and degree of unsaturation of the acyl groups. The developed method could resolve 18 lipid classes and multiple lipids within each class, from blood serum and brain tissue in 11 min.

七个亚2μm颗粒填充柱的性能（2-甲基吡啶胺，2-PIC;带电表面杂化氟苯基，CSH-FP;高强度硅胶C18 SB，HSS-C ₁₈ ;二乙胺，DEA; 1-氨基蒽，1-AA；高密度二醇和乙烯桥联的杂化物； BEH）在超高效超临界流体色谱（UHPSFC）和四极杆飞行时间质谱联用中进行了脂质分离研究。基于柱筛选的结果，开发了一种从主要脂质类别中分析多种脂质种类的方法。

含有β-羟基胺的固定相（即1-AA，DEA和2-PIC）具有较强的保留能力，并且具有伯胺基的两性离子脂质（如磷脂酰丝氨酸，磷脂酰乙醇胺及其溶菌形式）的峰形差。BEH和HSS-C ₁₈色谱柱分别显示了极性脂质和非极性脂质的强保留能力。Diol色谱柱保留了大多数主要脂质类别，并且还产生了对称峰。此外，该色谱柱还在主要脂质类别内和之间产生了最高的分离度。与乙酸铵或不存在添加剂相比，由甲醇：氯仿（1：2，v：v）和在流动相中添加20 mM甲酸铵组成的进样溶剂可改善色谱分离和质谱检测。最后，使用实验方法设计为Diol色谱柱优化了色谱和质谱参数。Diol色谱柱上的分离机理取决于脂质官能团以及酰基的长度和不饱和度。