Oral Sessions (Day1, Day2, Day3)
Oral Sessions
- Day 1, May 19（Wed.） 15:15-15:30 Room E (Zoom)
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1E-O5-1515 PDF
Metabolomics of Small Cell Populations by High-Sensitive Nano-uHILIC/AEX/MS/MS
The rapid development of next-generation sequencing techniques has enabled genomic and transcriptomic analyses at single-cell resolution, which have revealed the importance of minority cell populations in biological systems. However, analytical methods to accurately identify and quantify comprehensive metabolites from small cell population samples are still in the process of development. The aim of this study was to develop an analytical system of trace cell samples (< 100 cells) based on highly sensitive nano-unified hydrophilic-interaction/anion-exchange liquid chromatography quadrupole Orbitrap mass spectrometry (nano-uHILIC/AEX/MS/MS). A packed nano-LC column (3.5-µm particle size GL-HilicAex of dimensions 100 µm i.d. × 180 mm) was prepared using a slurry technique. The optimized nano-uHILIC/AEX/MS/MS method showed about 40-fold greater sensitivity in average than those in conventional semimicro-uHILIC/AEX/MS/MS. The acquisition was achieved in full scan and parallel reaction monitoring (PRM) mode. We successfully quantified 16 targeted hydrophilic metabolites and detected 373 compound features from two types of rare immune cells (6~50 cells, n = 13) using nano-uHILIC/AEX/MS/MS. We identified significant components by a volcano plot that suggest differences in mouse immune cell function. The developed analytical system represents a potentially useful tool for in-depth studies focused on small cell populations.