A novel, fast and sensitive supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) method for analysis of arachidonic acid metabolites

Abstract

The development of a rapid, sensitive and reliable method for the quantification of bioactive arachidonic acid metabolites (AA-metabolites) in biological samples is quite challenging due to the minute concentration, short half-life and their structural complexity arising from different isomers. In this study, a simple, fast and environmentally friendly supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) method was developed and validated for simultaneous measurement of five (PGD2, PGE2, PGF2α, 6KetoPGF1α and LTB4) AA-metabolites in biological samples. These analytes were extracted by protein precipitation followed by separation and quantification. The analysis was completed within 3 minutes. The matrix matched linear calibration ranged from 0.5-100 ng mL-1 (r2 ≥ 0.995), whilst, the limit of quantification of PGD2, PGE2, PGF2α, and LTB4 was 0.5 ng mL-1 and was 2.5 ng mL-1 for 6KetoPGF1α. The interday and intraday precisions of the method were less than 15% while the accuracy of most of the analytes varied between 83 and 109%. Finally, as a proof of concept, the method was successfully applied for the determination of eicosanoids in human samples, which expands the possibility to explore physiological states, disease phenotypes, and novel biomarkers.

Development of a Chiral Supercritical Fluid Chromatography–Tandem Mass Spectrometry and Reversed-Phase Liquid Chromatography–Tandem Mass Spectrometry Platform for the Quantitative Metabolic Profiling of Octadecanoid Oxylipins

“…The nature of the matrix effect measured by SFC-MS, however, depended upon the employed stationary phase. To our knowledge, no evaluation of the matrix effects on the column employed in this study has been reported, but a method for the analysis of plasma eicosanoids in achiral SFC showed similar signal enhancements …”

“…To our knowledge, no evaluation of the matrix effects on the column employed in this study has been reported, but a method for the analysis of plasma eicosanoids in achiral SFC showed similar signal enhancements. 23 Recoveries were evaluated at three concentration levels and were 30−120% for all analytes in SFC (average values: 74, 80, and 77%, Table S14) and between 13 and 110% in LC (average values: 83, 75, and 71%, Table S15), where the only species with a recovery of <30% was 11(E)-10-HOME. Recoveries for PhytoPs were calculated only by LC and were acceptable for all species (83−106%).…”

Analysis of Nonvolatile Molecules in Supercritical Carbon Dioxide Using Proton-Transfer-Reaction Ionization Time-of-Flight Mass Spectrometry

“…Mass spectrometry (MS) has also been applied to SFC for some time. − MS has a higher capability for molecule identification than a UV diode-array detector. The mobile phase CO 2 has fewer chemical background signals than the commonly used organic solvents, which is expected to decrease the limit of detection (LOD).…”

Supercritical fluid chromatography coupled to mass spectrometry for lipidomics

“…Targeted lipidomics methods in SFC-MS have been focused on several classes such as free fatty acids (C18 column, methanol/acetonitrile (50:50, v/v) with 0.1% formic acid) 35 or vitamin E derivatives in vegetable oils (NH2 column, pure ethanol), 27 carotenoids in human plasma (C30 column, pure methanol), eicosanoids in human plasma and cerebrospinal fluid (2picolamine column, MeOH with 0.1% formic acid and 10 mM of ammonium acetate), 36 steroids (2-ethylpyridine column, pure isopropanol), 37 environmental toxins as acetogenins (2ethylpyridine column, pure ethanol). 28 These examples perfectly demonstrate the diversity of stationary phase and co-solvent that are described in the literature and which contrast strongly with HPLC studies that lead mainly to the use of C18 columns and water/acetonitrile gradients.…”