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A metabolomics platform by capillary electrophoresis coupled with a high-resolution mass spectrometry for plasma analysis Kazunori Sasaki, Hitoshi Sagawa, Makoto Suzuki, Hiroyuki Yamamoto, Masaru Tomita, Tomoyoshi Soga, and Yoshiaki Ohashi Anal. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.analchem.8b02994 • Publication Date (Web): 30 Nov 2018 Downloaded from http://pubs.acs.org on November 30, 2018
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Analytical Chemistry
A metabolomics platform by capillary electrophoresis coupled with a high-resolution mass spectrometry for plasma analysis Kazunori Sasaki1,2‡, Hitoshi Sagawa1‡, Makoto Suzuki1‡, Hiroyuki Yamamoto1, Masaru Tomita2, Tomoyoshi Soga2, Yoshiaki Ohashi1* 1Human
Metabolome Technologies Inc., 246-2 Mizukami, Kakuganji, Tsuruoka, Yamagata 997-0052, Japan for Advanced Biosciences, Keio University, 246-2 Mizukami, Kakuganji, Tsuruoka, Yamagata 997-0052, Japan
2Institute
ABSTRACT: Metabolome analysis using capillary electrophoresis (CE) coupled with high-resolution mass spectrometry (HRMS) has the potential to improve coverage of metabolite detection due to its high selectivity and sensitivity. Configuration of the interface between CE and HRMS to meet the ground connection is essential to enable independent regulation of the electrical currents in the CE and electrospray field. In the present study, we applied an electrospray ionization adapter equipped with a grounded nebulizer to CE-HRMS and tested the analytical performance for 34 charged compounds. The extracted ion electropherograms, consisting of seven sets of isomers, showed reasonable peak shapes and separation for the annotation of each metabolite. The levels of 34 target analytes in a standard mixture were determined with a dynamic range of at least 102, maintaining linearity with r2 >0.9. The repeatability and intermediate precision above the lower limit of quantification showed the relative standard deviation to be lower than 20%. In the spike recovery experiment, 27 of the 34 metabolites in plasma extract were recovered at a rate of 80 to 120%, suggesting high accuracy. Furthermore, we assessed the feasibility of our platform to metabolome analysis using human plasma extract. The results showed successful detection of 270 metabolites, indicating the potential of our platform to yield higher coverage of the metabolome. In addition, analysis of dilution integrity demonstrated the quantitative ability of metabolome analysis with CEHRMS, although the existence of saturation or matrix effects were seen in the case of 33 of the metabolites. This study indicates that our platform has great potential for large scale metabolome analysis of plasma for biological studies and clinical biomarker screening.
Biomarker discovery is driven by advanced techniques with the ability to detect changes in the presence or relative levels of various molecular species associated with disease. Metabolomics is a large-scale approach to obtain comprehensive biological information about low molecular weight metabolites (0.9. The boundaries of the linear range varied among the analytes investigated due to differences in the ionization efficiency. The obtained LLOD values were in the nanomolar range. Although the use of HRMS can improve signal-to-noise ratios, the detection sensitivity might be compromised due to dilution of the capillary effluent by the sheath liquid. The accuracy of the measured concentration of the standards indicated that the peak signals of highly abundant analytes may become saturated, resulting in decreased accuracy in quantification. Linearity of the calibration curves was maintained for analyte concentrations up to 100 µM. The results of repeatability and intermediate precision measurements of analyte concentration showed acceptable performance above the LLOQ, with RSD lower than 20% for quantitative analysis. These measurements reflect the instrument variability, which can affect the quantification of analytes. The results of the assessment of metabolite quantitativity are shown in Table S4. The levels of most metabolites were measured with good precision, with RSD
