Highly Sensitive Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry for High-Throughput Metabolic Profiling

摘要

In the present study, a high-throughput and nontargetednmetabolomic technique using matrix-assisted laser de-nsorption ionization-mass spectrometry (MALDI-MS) wasndeveloped for the rapid analysis of cellular metabolites.nEither the detection limit or the linearity between con-ncentrations of several standards and peak intensities wasnexamined, indicating a detection limit lower than 10 fmolwell with a high linearity at low concentrations. To verifynthe validity of this method, metabolites from human acutenlymphoblastic leukemia Jurkat cells were analyzed. Onlyn2 500 cells suspended in PBS were directly dropped ontona stainless MALDI sample plate, followed by mixing withnmatrix on the sample plate. Up to 150 metabolite peaksnwere detected from a single analysis within 90 s. Fornmultivariate analysis of Jurkat cells against drug-treat-nment, three anticancer drugs were utilized. Principalncomponent analysis of metabolites showed clear indepen-ndent clusters for cells treated with these anticancer drugs.nFurthermore, several metabolites involved in nucleotidensynthesis were found to contribute to the separation ofneach cluster. These data suggest that the high-throughputnMALDI-MS-based metabolomic technique proposed in thenpresent study can be utilized for drug screening andnvalidation of drug efficacy and safety.