3A-O1-0940 PDF
Development of an ultra-high sensitive method for targeted lipidomics by LC-MS/MS in biological samples
Metabolomics is one of “Omics” approaches for researching a clinical biomarker of disease prediction and progression. Recently, lipid mediators, such as lysophospholipids (LPLs) have been focused in human metabolism. Recently growth of LPLs analysis has greatly been depended on the improvement of mass spectrometry (MS), especially for “untargeted” lipidomics. However, the reproducibility and sensitivity of analysis are still developing. Therefore, we have focused on the LC separation and developed a“targeted”method of 50 LPLs using LC-MS/MS. The LC separation was performed using a Capcell Pak ACR (150 mm x 1.5 mm i.d., 3 μm particle size) with a gradient elution of solvent A (5 mmol/L ammonium formate in water, pH 4.0) and solvent B (5 mmol/L ammonium formate in 95% acetonitrile, pH 4.0) at 150 μL/min. The lower limit of quantification is under 10 pmol/L for LPLs. We evaluated the method and obtained good values of accuracy (~15.0%) and precision (~10.0%). Finally, we also have developed an ultra-high sensitive method coupled with a column switching, and LPLs were detected in micro-volume of plasma (~0.1 μL) or micro-tissue (~100 cell). The present method of“targeted”lipid analysis is effective for validating lipid biomarkers.