Evaluation of Two Simultaneous Metabolomic and Proteomic Extraction Protocols Assessed by Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry

Untargeted multi-omics analysis of plasma is an emerging tool for the identification of novel biomarkers for evaluating disease prognosis, and for developing a better understanding of molecular mechanisms underlying human disease. The successful application of metabolomic and proteomic approaches re...

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Published in:International journal of molecular sciences 2023-01, Vol.24 (2), p.1354
Main Authors: Zenati, Ruba A, Giddey, Alexander D, Al-Hroub, Hamza M, Hagyousif, Yousra A, El-Huneidi, Waseem, Bustanji, Yasser, Abu-Gharbieh, Eman, Alqudah, Mohammad A Y, Shara, Mohd, Abuhelwa, Ahmad Y, Soares, Nelson C, Semreen, Mohammad H
Format: Article
Language:English
Subjects:
Biological variation
Blood plasma
Chemical precipitation
Chlorine compounds
Chloroform
Chromatography, High Pressure Liquid - methods
Disease
Glycerol
High performance liquid chromatography
Humans
Mass spectrometry
Mass spectroscopy
Metabolism
Metabolites
Metabolomics
Methanol - chemistry
Methods
Molecular modelling
multi-omics
Peptides
Plasma
Proteins
Proteomics
Quadrupoles
Reproducibility
Reproducibility of Results
Solvents
Spectroscopy
Tandem Mass Spectrometry - methods
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Summary:Untargeted multi-omics analysis of plasma is an emerging tool for the identification of novel biomarkers for evaluating disease prognosis, and for developing a better understanding of molecular mechanisms underlying human disease. The successful application of metabolomic and proteomic approaches relies on reproducibly quantifying a wide range of metabolites and proteins. Herein, we report the results of untargeted metabolomic and proteomic analyses from blood plasma samples following analyte extraction by two frequently-used solvent systems: chloroform/methanol and methanol-only. Whole blood samples were collected from participants (n = 6) at University Hospital Sharjah (UHS) hospital, then plasma was separated and extracted by two methods: (i) methanol precipitation and (ii) 4:3 methanol:chloroform extraction. The coverage and reproducibility of the two methods were assessed by ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). The study revealed that metabolite extraction by methanol-only showed greater reproducibility for both metabolomic and proteomic quantifications than did methanol/chloroform, while yielding similar peptide coverage. However, coverage of extracted metabolites was higher with the methanol/chloroform precipitation.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms24021354