title: A reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling.
abstract: Lipidomics is a critical part of metabolomics and aims to study all the lipids within a living system. We present here the development and evaluation of a sensitive capillary UPLC-MS method for comprehensive top-down/bottom-up lipid profiling. Three different stationary phases were evaluated in terms of peak capacity, linearity, reproducibility, and limit of quantification (LOQ) using a mixture of lipid standards representative of the lipidome. The relative standard deviations of the retention times and peak abundances of the lipid standards were 0.29% and 7.7%, respectively, when using the optimized method. The linearity was acceptable at >0.99 over 3 orders of magnitude, and the LOQs were sub-fmol. To demonstrate the performance of the method in the analysis of complex samples, we analyzed lipids extracted from a human cell line, rat plasma, and a model human skin tissue, identifying 446, 444, and 370 unique lipids, respectively. Overall, the method provided either higher coverage of the lipidome, greater measurement sensitivity, or both, when compared to other approaches of global, untargeted lipid profiling based on chromatography coupled with MS.
authors.affiliation: Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Gao
authors.forename: Qibin
authors.initials: Q
authors.lastname: Zhang
authors.forename: Da
authors.initials: D
authors.lastname: Meng
authors.forename: Giorgis
authors.initials: G
authors.lastname: Isaac
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Fillmore
authors.forename: Rosey K
authors.initials: RK
authors.lastname: Chu
authors.forename: Jianying
authors.initials: J
authors.lastname: Zhou
authors.forename: Keqi
authors.initials: K
authors.lastname: Tang
authors.forename: Zeping
authors.initials: Z
authors.lastname: Hu
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Moore
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
authors.forename: Michael G
authors.initials: MG
authors.lastname: Katze
authors.forename: Thomas O
authors.initials: TO
authors.lastname: Metz
mesh_terms: D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D008055:Lipids / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods*; D051381:Rats
chemical_list: D008055:Lipids