Thin-layer chromatography and coomassie staining of phospholipids for fast and simple lipidomics sample preparation

Abstract

Lipids play major roles in basic cellular functions. Their analysis, therefore, gained importance; however, it is complicated by the high complexity of natural lipidomes. To overcome this challenge, pre-separation of the lipids by thin-layer chromatography (TLC) and high-resolution mass spectrometry (MS) are often employed. Here, we explore Coomassie staining for TLC-separated phospholipids and provide an extraction protocol of stained lipids for subsequent MS analysis. We exemplify our approach by analyzing lipid mixtures varying in complexity and found that TLC pre-separation increases the number of identified lipid species and lipid classes. In addition, we identify and quantify lipids from polymer nanodiscs. In summary, Coomassie staining of TLC-separated lipids is well-suited for phospholipids, is compatible with MS, does not require specialized equipment and can be performed independently of subsequent MS experiments. We envision many future applications of our workflow.