Simultaneous profiling of lysophospholipids and phospholipids from human plasma by nanoflow liquid chromatography-tandem mass spectrometry

Abstract

In this study, an analytical method for the simultaneous separation and characterization of various molecular species of lysophospholipids (LPLs) and phospholipids (PLs) is introduced by employing nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (nLC-ESI-MS/MS). Since LPLs and PLs in human plasma are potential biomarkers for cancer, development of a sophisticated analytical method for the simultaneous profiling of these molecules is important. Standard species of LPLs and PLs were examined to establish a separation condition using a capillary LC column followed by MS scans and data-dependent collision-induced dissociation (CID) analysis for structural identification. With nLC-ESI-MS/MS, regioisomers of each category of LPLs were completely separated and identified with characteristic CID spectra. It was applied to the comprehensive profiling of LPLs and PLs from a human blood plasma sample and yielded identifications of 50 LPLs (each regioisomer pair of 6 lysophosphatidylcholines (LPCs), 7 lysophosphatidylethanolamines (LPEs), 9 lysophosphatidic acid (LPAs), 2 lysophosphatidylglycerols (LPGs), and 1 lysophosphatidylserine (LPS)) and 62 PLs (19 phosphatidylcholines (PCs), 11 phosphatidylethanolamines (PEs), 3 phosphatidylserines (PSs), 16 phosphatidylinositols (PIs), 8 phosphatidylglycerols (PGs), and 5 phosphatidic acids (PAs)).

Rapid tin-mediated access to a lysophosphatidylethanolamine (LPE) library: Application to positional LC/MS analysis for hepatic LPEs in non-alcoholic steatohepatitis model mice

“…Although three olefinic isomers of C18:3 (12-14) could not be separable in this condition, other LPEs were possible. Regioisomers can be distinguished by the LC and MS/MS because similar fragmentation patterns are observed as reported (Fang, et al, 2003;Lee, et al, 2011 (Figure 3 and SI).…”

“…Conversely, mass spectrometry requires very small sample quantities, which renders solubility virtually irrelevant. Especially liquid chromatography tandem-mass spectrometry (LC-MS/MS) represents a powerful approach to distinguish 1/2-LPEs (Han and Gross, 1996;Fang, et al, 2003;Lee, et al, 2011).…”

Rapid Screening of Phospholipid Biomarker Candidates from Prostate Cancer Urine Samples by Multiple Reaction Monitoring of UPLC-ESI-MS/MS and Statistical Approaches

“…13,14 However, liquid chromatography-ESI-tandem mass spectrometry (LC-ESI-MS/MS) in particular facilitates lipidomic analysis in separating complicated lipid mixtures with subsequent soft ionization of lipid molecules without preliminary modification and in deriving structural information from fragments of target ions. [15][16][17] This study investigated potential urinary phospholipid biomarkers of prostate cancer using ultrahigh performance LC-ESI-MS/MS (UPLC-ESI-MS/MS) with statistical analysis. Initially, 52 urinary PL species, found greater than the limit of quantitation of method applied in present study and identified by structural determination using nanoflow LC/ ESI-MS/MS in the previous study 25 were selected for the high speed screening of a large number of urine samples with prostate cancer.…”

Effects of Column Length and Particle Diameter on Phospholipid Analysis by Nanoflow Liquid Chromatography-Electrospray Ionization-Mass Spectrometry

“…10 Recently, reversed phase nanoflow LC-ESI-MS-MS has shown its capability in simultaneous separation and structural identification of complicated PL and LPL mixtures from human urine or blood plasma samples. 11,12,13,14,15 It was reported that the limit of detection (LOD) can be lowered to 2.2 fmol of phosphatidylcholine (PC) by nLC-ESI-MS experiments.…”

“…The detailed packing procedure for a pulled tip capillary column can be found from previous reports. 13,14,15 For nLC-ESI-MS-MS experiments, a model 1200 pump system from Agilent Technologies (Palo Alto, CA, USA) equipped with an autosampler and an LTQ Velos ion trap mass spectrometer (Thermo Finnigan, San Jose, CA, USA) were utilized. The capillary column was connected with pump via a PEEK MicroCross (Upchurch Scientific, Oak Harbor, WA, USA) in which capillary tubes from the pump, capillary column, a Pt wire for ESI voltage, and vent tube were all connected.…”