Determination of lipid mediators in breast cancer cells using lyophilization‒supercritical fluid extraction online coupled with supercritical fluid chromatography‒quadrupole tandem mass spectrometry

Abstract

A lyophilization-supercritical fluid extraction coupled with supercritical fluid chromatography-quadrupole tandem mass spectrometry online method was developed for the determination of lipid mediators in breast cancer cells. Supercritical fluid extraction was applied to the cell samples for the first time due to the use of lyophilization. The conditions of supercritical fluid extraction and supercritical fluid chromatography-quadrupole tandem mass spectrometry were investigated systematically. Under the optimized conditions, all the calibration curves for the lipid mediators showed good linearity (correlation coefficient > 0.99). The limits of detection and the limits of quantification were in the range of 0.190-5.36 pg and 0.560-16.2 pg, respectively. The recoveries were in the range of 70.3-125%. The relative standard deviations of the precision ranged from 1.49-18.7% and the accuracies were higher than 84%. Compared with liquid-liquid extraction coupled with liquid chromatography and tandem mass spectrometry method, the present approach reduced the manual labor and obtained higher sensitivity as well as higher extraction recoveries for all 15 lipid mediators. Finally, the online method was applied to the quantification of lipid mediators in breast cancer cells and normal mammary epithelial cells. On the basis of the results, this lyophilization-supercritical fluid extraction online coupled with supercritical fluid chromatography-quadrupole tandem mass spectrometry method showed great promise in the analysis of lipid mediators in complex biological samples.

Development of SFC-MS Method for Quantification of Eicosanoids Biosynthesized in Primary Human Blood Cells

“…Supercritical CO 2 is non-polar, but by the addition of varying amounts of polar modifiers such as methanol (MeOH) or isopropanol (IPA), separation of both polar and non-polar analytes is facilitated, which suggests SFC for analyzing oxidized polyunsaturated fatty acids ranging in polarity and occurring as stereoisomers. However, despite the development of robust instrumentations, so far only a few reports have been found that utilize SFC for oxylipin detection and quantification [15][16][17][18]. Here, we established and validated an SFC method coupled to a single quadrupole mass spectrometer to identify and quantify relevant eicosanoids.…”