The dispersive micro-solid phase extraction method for MS-based lipidomics of human breast milk

•Dispersive micro-solid phase extraction (D-μ-SPE) technique for lipidomics.•Combining stationary phase for extended lipidome coverage.•Application of 2k full factorial design for screening the factors affecting the D-μ-SPE. A simple and rapid microextraction method ensuring high lipidome coverage was developed for liquid chromatography mass spectrometry (LC-MS)-based lipidomics of human breast milk. The dispersive microsolid phase extraction (D-µ-SPE) technique, coupled with the design of experiment (DoE) method, enabled the study of the influence of several conditions (desorption solvent, stationary phase ratio, and sorption and desorption time) on the lipid extraction process of various lipid classes. The D-µ-SPE-based method, which used a mixture of C18 and zirconia-coated silica gel as the sorbent, allowed for the extraction of a wide range lipid classes characterized by different concentration levels. The developed method simplified the extraction procedure for lipidomics without loss of good reproducibility (70% of the MFs had peak volume%RSD