Lipidomic analysis of skeletal muscle tissues of p53 knockout mice by nUPLC-ESI-MS/MS

Abstract

Tumour suppressor p53 is known to be associated with the maintenance of mitochondrial functional properties in the skeletal muscles. As deactivation or mutation of p53 can affect the synthesis of lipids, investigating the relationship between p53-related energy generation metabolism and perturbation of lipid profile is critical. In this study, 329 lipid species (among 412 identified species) in two different skeletal muscle tissues (the gastrocnemius and soleus) from p53 knockout (KO) mice were quantitatively analysed using nanoflow ultrahigh performance liquid chromatography tandem mass spectrometry (nUPLC-MS/MS). Overall, lipids from the soleus tissues were more affected by p53 KO than those from the gastrocnemius in most lipid profiles. In p53 KO, lysophosphatidylcholine (LPC), lysophosphatidylserine (LPS), phosphatidic acid (PA), sphingomyelin (SM), and triacylglycerol (TAG), including 6 TAG (44:2, 46:0, 58:5, 58:8, 58:9, and 50:0), were significantly increased (p < 0.05) by 1.4–2-fold only in the soleus tissue. Overall monohexosylceramide (MHC) levels, including those of 3 MHC species (d18:0/24:0, d18:1/22:0, and d18:1/24:0), were significantly increased (p < 0.05) by 2–4 fold, only in the gastrocnemius tissue. The results suggest that lipid profiles are significantly altered by the lack of p53 in muscle tissues.

Nano-LC/NSI MS Refines Lipidomics by Enhancing Lipid Coverage, Measurement Sensitivity, and Linear Dynamic Range

“…Notably, also other researchers pursued a miniaturization of the chromatography for lipid analysis pioneering from Taguchi et al (2000) and Bang et al (2006) . Gao et al (2012) were able to describe 238 species level phospholipids (446 species level lipids) in a human cell line, while Park et al (2017) analyzed 169 (mixed species level and molecular species level) phospholipids (412 total lipids) in skeletal muscle tissue. Hence, this study is likely setting a new benchmark for confident comprehensive phospholipid data sets.…”

“…All in all, nLC/NSI systems provide a strategy to come closer toward the goal of comprehensive lipidome coverage even when sample availability is limited. However, despite the apparent advantages of nLC-MS systems for lipid analyses, this promising technology has not yet been established in the lipidomics community even though several laboratories already applied nano/capillary chromatographic systems for (often class restricted) lipid analyses. ,− Nanoscale chromatography for lipidomics appears only to be routinely used by the Moon lab so far. ,,− …”

Enhanced Phospholipid Isomer Analysis by Online Photochemical Derivatization and RPLC-MS

“…Other common RPLC stationary phases have also been reported for phospholipid separation, such as C8 and C30. ,, The choices of mobile phase for RPLC of phospholipids are very limited. Isopropanol/acetonitrile (90/10, v/v) and acetonitrile/water (60/40, v/v) are used as the standard mobile phase in most phospholipid separations, while incorporation of methanol into these solvents was also reported. , To obtain adequate elution of lipids, isopropanol is usually required as the strong elution solvent. Relatively higher column temperatures (e.g., 50–60 °C) are usually used to further help desorption of lipids.…”

Lipidomics Profiling of HepG2 Cells and Interference by Mycotoxins Based on UPLC-TOF-IMS

“…13−18 For example, gas chromatography−mass spectrometry shows excellent resolution ability in fatty acid analysis 19,20 and liquid chromatography is often coupled online with ion trap, triple-stage quadrupole, Q-Exactive Orbitrap mass spectrometry, etc. 21,22 Nevertheless, these techniques are limited by factors including matrix effects and interference of isomers and isobars created by the structural diversity of metabolites and lipids, which can confound identification. Newer innovations, such as IMS, are gaining popularity for the analysis of lipids, where large numbers of ions have the same mass values arising from different headgroups, different connectivities of fatty acyls, and different locations of double bonds within the fatty acyls.…”

“…There is no doubt that hyphenated and multidimensional techniques based on mass spectrometry are currently the most extensive and effective tools for lipid/metabolite profiling and quantification. − For example, gas chromatography–mass spectrometry shows excellent resolution ability in fatty acid analysis , and liquid chromatography is often coupled online with ion trap, triple-stage quadrupole, Q-Exactive Orbitrap mass spectrometry, etc. , Nevertheless, these techniques are limited by factors including matrix effects and interference of isomers and isobars created by the structural diversity of metabolites and lipids, which can confound identification. Newer innovations, such as IMS, are gaining popularity for the analysis of lipids, where large numbers of ions have the same mass values arising from different headgroups, different connectivities of fatty acyls, and different locations of double bonds within the fatty acyls. , Reduced mobilities and collision cross sections (CCSs), which are determined for lipid identification by comparison with lipid databases, are more reproducible than the retention time (RT) due to the potential for RT shifts and have been used to determine the degree of isomerism (e.g., different double bond positions, geometries) in untargeted lipidomics .…”