Comprehensive phospholipid and sphingomyelin profiling of different brain regions in mouse model of anxiety disorder using online two-dimensional (HILIC/RP)-LC/MS method

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 149; pp. 308 - 317
• Main Authors: Berkecz, Róbert, Tömösi, Ferenc, Körmöczi, Tímea, Szegedi, Viktor, Horváth, János, Janáky, Tamás
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 05.02.2018
• Subjects: 2D-LC/MS; Animals; Anxiety disorder; Anxiety Disorders - pathology; Brain; Brain - pathology; Brain Chemistry; Chromatography, Reverse-Phase - instrumentation; Chromatography, Reverse-Phase - methods; Disease Models, Animal; Humans; Hydrophobic and Hydrophilic Interactions; Lipidomics; Male; Mice; Mice, Inbred Strains; Phospholipid; Phospholipids - analysis; Pilot Projects; Spectrometry, Mass, Electrospray Ionization - instrumentation; Spectrometry, Mass, Electrospray Ionization - methods; Sphingomyelin; Sphingomyelins - analysis; RS; Brain; NP; PS; Anxiety disorder; Phospholipid; LPL; PFC; VHPC; DHPC; ME; SM; AGC; HILIC; nAX; OPLS-DA; Lipidomics; IS; C18; PA; 1D; PC; RE; PE; AX; PG; Sphingomyelin; 2D-LC/MS; PI; SEM; FA; PL; RP
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2017.10.043

•Development of novel online 2D-LC/MS method for comprehensive analysis of phospholipids and sphingomyelins in brain sample.•Profiling of PL and SM composition of DHPC, VHPC and PFC brain regions was achieved in mouse model of anxiety disorder.•In case of 37 PL and SM species, significant differences were observed between AX and nAX mice. A novel online system including two-dimensional liquid chromatography coupled to high-resolution mass spectrometry (2D-LC/MS) was developed and applied for comprehensive phospholipid (PL) and sphingomyelin (SM) profiling of dorsal hippocampus (DHPC), ventral (VHPC) and prefrontal cortex (PFC) brain regions in a mouse model of anxiety disorder. In the first dimension, lipid classes were distinguished by hydrophilic interaction liquid chromatography (HILIC), while the second dimensional separation of individual PL and SM species was achieved by reversed-phase (RP) chromatography. For the enrichment of lipid species in diluted HILIC effluent, two RP trapping columns were used separately. The developed fully-automated 2D method allowed the quantitative analysis of over 150 endogenous PL and SM species in mouse brain regions within 40min. The developed method was applied in a pilot study, which aimed to find alteration of PL and SM composition in a mouse model of anxiety disorder. In the case of 37 PL and SM species, significant differences were observed between high anxiety-related behavior (AX) and low anxiety-related behavior (nAX) mice. In mice having elevated anxiety, the most typical trend was the downregulation of PL species, in particular, in VHPC.