IntroductionPreviously, we reported a method to profile regulatory lipid mediators, which play important roles in various biological functions such as angiogenesis, proliferation, apoptosis and etc., using scheduled MRM LC-ESI MS/MS method. In 21.5 minutes, 87 regulatory lipid mediators were measured simultaneously.  However, the sample preparation method-SPE method is labor extensive and time consuming. Here, we report a method for profile the regulatory lipid mediators using 96 well plate format SPE and Phree plate to provide the higher throughput than the original method.Methods:The regulatory lipid mediators' standards (including 87 lipid mediator and 9 deterated internal standards) spiked into human plasma at various concentrations (3 nM, 30 nM and 300 nM). The samples were extracted through 96 well plate format of Oasis HLB uElute 96 well plates (Waters) or Phree Phospholipid Removal 96-well plate (Phenomenex). The extraction eluents were mixed with loading internal standards in a 96-well storage plate before loading on the LC/MS/MS for the separation and detection.Preliminary data:There is significant improvement for the throughput compared to original method (4 plates x 96 than 40 SPE cartridge per day). There is no significant difference regarding on the recovery rate of the extraction between the 96 well plates and cartridge format. The reproducibility of the new methods are 80-120%.Novel Aspects:Application of 96-well plate extraction method allows increasing the throughput of regulatory lipid mediators profiling dramatically.

Application of 96 well plate solid phase extraction to increase the throughput of regulatory lipid mediators metabolomic profiling in plasma

248th National Meeting (2014)

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