A Simple and Highly Sensitive Quantitation of Eicosanoids in Biological Samples Using Nano-flow Liquid Chromatography/ Mass Spectrometry

Abstract

A simple sample preparation method for eicosanoid was developed by the combination of deproteinization and nanoLC-ESI-MS/MS. Eicosanoids are a group of bioactive lipid mediators, present in trace amounts in the body. Therefore, an analytical method for eicosanoids requires superior sensitivity. The method described in this report, which takes advantage of the highly sensitive power of nanoLC-ESI-MS/MS, enabled a simplification of the sample-preparation process. Eicosanoid extraction was performed just by homogenization in methanol with subsequent phospholipid removal, and then the liquid phase was directly subjected to nanoLC-ESI-MS/MS analysis without a condensation process. The quantitation range achieved 0.01 -100 ng/mL for thromboxane B2, and 0.05 -100 ng/mL for prostaglandin E2, prostaglandin D2, prostaglandin F2, leukotriene B4, 6-keto prostaglandin F1α and 11-dehydro thromboxane B2. Rat brain sample analyses demonstrated the feasibility of the quantification of those seven eicosanoids from biological samples.

Untargeted Lipidomic Analysis of Plasma from High-fat Diet-induced Obese Rats Using UHPLC–Linear Trap Quadrupole–Orbitrap MS

“…In clinical chemistry, blood-based lipid biomarkers for obesity are crucial for the diagnosis of the disease. Moreover advanced LC/MS techniques were widely applied for lipid biomarker discovery in various models 26,27 . In the present study, a rodent rat model was used, and rats were fed with ND and HFD (fat source: HF-L-lard) for 8 weeks.…”

Targeted analysis of eicosanoids derived from cytochrome P450 pathway by high‐resolution multiple‐reaction monitoring mass spectrometry

“…Another advantage of the developed method is the simultaneous evaluation of a wide range of metabolites from CYP450, a poorly explored pathway. Many studies are focused primarily on COX and LOX pathway 34–36 . However, the balance between the CYP450 pathway and these other vias may be decisive for the global understanding of a biological phenomenon or elucidating a specific phenomenon in an immunological condition.…”

Miniaturized LC in Molecular Omics

“…In particular, LOQ values in a range of 10–50 ng/L have been reported for eicosanoid analysis using nano-LC–MS/MS in different biological tissues . Ando and Satomi demonstrated that the SP procedure can be minimized due to the higher sensitivity of nano-LC–MS/MS compared to other LC–MS/MS platforms (HPLC or even UHPLC) …”