Metabolomics Platform with Capillary Electrophoresis Coupled with High-Resolution Mass Spectrometry for Plasma Analysis

Abstract

Metabolome analysis using capillary electrophoresis (CE) coupled with high-resolution mass spectrometry (HRMS) has the potential to improve coverage of metabolite detection because of its high selectivity and sensitivity. Configuration of the interface between CE and HRMS to meet the ground connection is essential for enabling 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 10 2 , maintaining linearity with r 2 > 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 in 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 CE-HRMS, 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 largescale metabolome analysis of plasma for biological studies and clinical biomarker screening.

Dysregulation of multiple metabolic networks related to brain transmethylation and polyamine pathways in Alzheimer disease: A targeted metabolomic and transcriptomic study

“…Quantification is performed using the (M+H)+ or (M-H)− parent ion peak area for each metabolite compared to the same parent peak in the standard solution. Concentration is only reported if the measured area is above a signal-to-noise ratio (S/N) of 5 and the peak area is within the linear range of the standard curve [39]. Coefficients of variation (CVs) for repeat technical samples using this methodology were less than 10%, as reported previously by Soga and colleagues [40].…”

“…HMT's in-house metabolite library contains 1,363 compounds, as described recently by Sasaki and colleagues [39]. A full listing of available compounds in the library is accessible through their recent publication at https://pubs.acs.org/doi/abs/10.1021/acs.analchem.…”

Blood Metabolite Signature of Metabolic Syndrome Implicates Alterations in Amino Acid Metabolism: Findings from the Baltimore Longitudinal Study of Aging (BLSA) and the Tsuruoka Metabolomics Cohort Study (TMCS)

“…Quantification was performed using the (M+H) + or (M-H) − parent ion peak area for each metabolite compared to the same parent peak in the standard solution. Concentration is only reported if the measured area is above the signal:noise ratio (S/N) of 5 and peak area is within linear range of the standard curve [87]. As reported previously [88], coefficients of variation (CVs) for repeat technical was less than 10%.…”

Emerging Applications of Metabolomics in Clinical Pharmacology

“…In particular, the method is ultrasensitive and can detect ~60 metabolites from only 10,000 hematopoietic stem cells, which enables metabolomics profiling of rare cell populations . Recent studies have also utilized high resolution MS instruments such as Orbitrap and time‐of‐flight (TOF) for targeted metabolite quantification . For example, by using a quadrupole‐Orbitrap LC‐MS (Q‐Exactive, Thermo Scientific, Bremen, Germany), Zhou et al .…”

“…28 Recent studies have also utilized high resolution MS instruments such as Orbitrap and time-of-flight (TOF) for targeted metabolite quantification. 31 For example, by using a quadrupole-Orbitrap LC-MS (Q-Exactive, Thermo Scientific, Bremen, Germany), Zhou et al 32 developed an efficient largescale quantitative method targeting 237 metabolites by using the scheduled parallel reaction monitoring mode. In untargeted metabolomics, high resolution MS instruments such as quadrupole TOF and Orbitrap mass analyzers are extensively used because of their high mass resolution, high mass accuracy, and high scan speed.…”