In this study, protein precipitation and bioactive lipid extraction from rat skin tissues is reported. The combination Solid Phase Extraction and liquid chromatography tandem mass spectrometry (LCMS/ MS) approach was used to measure concentrations of lipid mediators in these tissues. Rat skin tissues were homogenized using CKMix50_7mL Precellys® Lysing Kit and Precellys® Evolution tissue homogenizer combined with Cryolys® cooling unit. In addition, temperatures of homogenates were measured to investigate if temperature of complete homogenate of rat skin tissues remained below 40°C.
Fig. 1 Images of a whole rat hindpaw tissue sample (a),
and homogenized tissue (b) using Precellys® Evolution.
Fig. 2 Bioactive lipid mediators of the Arachidonic acid
cascade were successfully measured, by LC-MS/MS, after
lipids were extracted from the homogenate by solid phase
extraction. Concentrations of 5-HETE, 8-HETE and 9-HETE
were measured to be 1.3 ±0.3, 7.9±0.6 and 1.3 ± 0.4
ng/mg of tissue, respectively.