日本質量分析学会 第69回質量分析総合討論会会

演題概要

オーラルセッション

第1日 5月19日(水) 15:15~15:30 E会場(Zoom)

高感度nano-uHILIC/AEX/MS/MSによる少数細胞集団試料のメタボローム分析

(1九大生医研2阪大院医)
o中谷航太1和泉自泰1秦康祐1高橋政友1内田穣2石井優2馬場健史1

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.