Highly sensitive and selective analysis of widely targeted metabolomics using gas chromatography/triple-quadrupole mass spectrometry

Abstract

In metabolomics studies, gas chromatography coupled with time-of-flight or quadrupole mass spectrometry has frequently been used for the non-targeted analysis of hydrophilic metabolites. However, because the analytical platform employs the deconvolution method to extract single-metabolite information from co-eluted peaks and background noise, the extracted peak is artificial product depending on the mathematical parameters and is not completely compatible with the pure component obtained by analyzing a standard compound. Moreover, it has insufficient ability for quantitative metabolomics. Therefore, highly sensitive and selective methods capable of pure peak extraction without any complicated mathematical techniques are needed. For this purpose, we have developed a novel analytical method using gas chromatography coupled with triple-quadrupole mass spectrometry (GC-QqQ/MS). We developed a selected reaction monitoring (SRM) method to analyze the trimethylsilyl derivatives of 110 metabolites, using electron ionization. This methodology enables us to utilize two complementary techniques-non-targeted and widely targeted metabolomics in the same sample preparation protocol, which would facilitate the formulation or verification of novel hypotheses in biological sciences. The GC-QqQ/MS analysis can accurately identify a metabolite using multichannel SRM transitions and intensity ratios in the analysis of living organisms. In addition, our methodology offers a wide dynamic range, high sensitivity, and highly reproducible metabolite profiles, which will contribute to the biomarker discoveries and quality evaluations in biology, medicine, and food sciences.

Investigations in the possibility of early detection of colorectal cancer by gas chromatography/triple-quadrupole mass spectrometry

“…QqQMS-based MRM analysis can be used to easily distinguish single metabolite-derived peaks from co-eluted peaks and background noise, and it exhibits high sensitivity and a wide dynamic range compared with single-QMS [9], although gas chromatography/single-quadrupole mass spectrometry (GC/QMS) is very useful for comprehensive metabolite analysis [10]. In our previous study [4], GC/QMS was used to discover biomarker candidates for colorectal cancer.…”

Molar-Based Targeted Metabolic Profiling of Cyanobacterial Strains with Potential for Biological Production

“…To perform widely targeted metabolic profiling in the multiple reaction monitoring (MRM) mode, gas chromatography–triple quadrupole mass spectrometry (GC/QqQ-MS) was used for analyzing amino acids and organic acids, while analysis of intermediates of central metabolism such as sugar phosphates and cofactors was performed using reserved phase ion-pair liquid chromatography (RP-IP-LC)/QqQ-MS. Triple quadrupole mass spectrometer is suitable for metabolic profiling because of its high sensitivity and selectivity as well as its wide dynamic range that makes it possible to give accurate quantitative information [22,23]. Moreover, to obtain the absolute concentration of each metabolite, we used 13 C-labeled cell extracts as internal standards, a technique known to effectively reduce errors due to variations occurring in the analysis and sample processing [24].…”

Technical Challenges in Mass Spectrometry-Based Metabolomics

“…37) Sawada et al reported widely targeted metabolome analysis using liquid chromatographtriple quadrupole-mass spectrometers (TQ-MS) by developing selected ion monitoring series of 378 metabolites. 38) Similar approaches are employed for the comprehensive analysis of lipids using liquid chromatograph-TQ-MS (lipidomics) 39) and analysis of primary metabolites using gas chromatograph-TQ-MS. 40) Although the number of measurable metabolites depends on the samples of interests, it has been reported that approximately 100-200 metabolites were usually analyzed by the widely targeted analysis. e widely targeted approach is now a productive and realistic strategy in metabolomics because the analysis could be performed using the more popular TQ-MS and default peak picking so ware developed by manufacturers.…”