Liquid chromatography – mass spectrometry (LC-MS)-based lipidomics is almost exclusively performed with analytical flow rates due to its throughput and robust operation. However, higher flow rates and larger column diameters lower the electrospray efficiency and hence sensitivity. To increase throughput and robustness of nanoflow separation, the Evosep One LC system elutes samples from disposable trap columns and embeds the analytes in a pre-formed gradient, which is then pushed through the separation column with a single high-pressure pump (PMID: 30104208). Here, we adapt this system to the specific challenges of lipidomics and evaluate its performance in combination with trapped ion mobility spectrometry (TIMS) and deep MS/MS analysis in single runs by parallel accumulation – serial fragmentation (PASEF, PMID: 31949144).