Evaluation of Two Simultaneous Metabolomic and Proteomic Extraction Protocols Assessed by Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry.
Ruba A ZenatiAlexander D GiddeyHamza M Al-HroubYousra A HagyousifWaseem El-HuneidiYasser K BustanjiEman Abu-GharbiehMohammad A Y AlqudahMohd SharaAhmad Y AbuhelwaNelson Cruz SoaresMohammad Harb SemreenPublished in: International journal of molecular sciences (2023)
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.
Keyphrases
- tandem mass spectrometry
- ultra high performance liquid chromatography
- ms ms
- liquid chromatography
- high resolution mass spectrometry
- high performance liquid chromatography
- mass spectrometry
- simultaneous determination
- carbon dioxide
- gas chromatography
- solid phase extraction
- high resolution
- label free
- endothelial cells
- multiple sclerosis
- healthcare
- gas chromatography mass spectrometry
- emergency department
- electronic health record
- drug induced