A versatile method to separate complex lipid mixtures using 1-butanol as eluent in a reverse-phase UHPLC-ESI-MS system

•1-butanol instead of 2-propanol as organic mobile phase in RPLC separations extends elution capabilities.•The 1-butanol based method allows for separation of a large range of lipids with different hydrophobicities.•Effective separation of complex biological lipid mixtures demonstrates the versatility of the method for lipidomic analyses.•A complex mixture of cardiolipins elutes in clusters dependent on the number of carbon atoms in the acyl chain.•Isobaric diester and cardiolipin lipid species separation allows for more detailed acyl chain analysis.