Development of an extraction method for the analysis of pro-inflammatory prostaglandin-E2 and leukotriene-B4 in human plasma by liquid chromatography tandem mass spectrometry

Abstract

A simple and rapid method for extracting PGE2 and LTB4 from human plasma and further determination by LC-MS/MS proposed and validated. Extracting solvents, formic acid (10 µL) and acetonitrile (140 µL) were added subsequently to 50 µL plasma sample. After vortex-mixing and centrifuging the obtained solution, the supernatant was submitted to LC-MS/MS. The quantitative analysis was carried out based on the internal standard method and the chromatographic separation using the LC column gradient mobile phase system. The analytical species were recorded by multiple reaction monitoring in negative mode. The method was validated using blank human plasma in the range of 1-50 ng/mL, and it exhibits good selectivity with LOD and LOQ of 0.4 ng/mL and 1 ng/mL respectively for PGE2 and LTB4 analytes. The recovery ranges were from 80.54 - 104.96% for PGE2 and 72.34 - 105.61% for LTB4. The method was applied to 40 human plasma samples from patients participating in clinical nutritional intervention studies and suffering from inflammatory disorders.