Determination of Inflammation-Related Lipids in Depressive Rats by On-Line Supercritical Fluid Extraction-Supercritical Fluid Chromatography-Tandem Mass Spectrometry

An on-line supercritical fluid extraction coupled with supercritical fluid chromatography-quadrupole tandem mass spectrometry method was developed to determine lipids related to inflammation in brain tissues of depressed rats. The analysis of 23 lipids from extraction to separation and detection only took 15 min and required 1 mg of brain tissue powder. The matrix effect of the on-line method for endogenous lipids was systematically investigated, and targeted lipids were quantified by matrix effect corrected calibration curves in this study. The on-line method was comprehensively optimized and evaluated. All calibration curves for lipids showed good linearity (correlation coefficient >0.99). The limits of detection and the limits of quantification were in the range of 0.0261–0.396 pg and 0.0791–1.20 pg. The recoveries and the matrix effect were in the range of 85.3–117.5% and 51.9–176.6%, respectively. The relative standard deviations of precision ranged from 2.7 to 14.2%, with accuracies higher than 87.2%. Compared with liquid-liquid extraction coupled with liquid chromatography-tandem mass spectrometry method, the on-line method obtained higher recovery and sensitivity with significantly reduced analytical time, manual operations, and sample amounts. Finally, this on-line method was applied to analyses of brain tissues of depressed rats. Six pro-inflammatory lipids increased in depressed rats, while six anti-inflammatory lipids decreased. Liquiritin and fluoxetine were presumed to promote a similar synthesis of anti-inflammatory lipids. Based on the results, this on-line method showed great promise in analyzing lipids in complex biological samples.