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A Protocol for Untargeted Metabolomic Analysis: From Sample Preparation to Data Processing.

Amanda L Souza¹, Gary J Patti^2,3

¹Life Science Mass Spectrometry Division, Thermo Fisher Scientific, San Jose, CA, USA. amanda.souza@thermofisher.com.

Methods in Molecular Biology (Clifton, N.J.)

|June 1, 2021

Summary

This summary is machine-generated.

This study presents a detailed protocol for untargeted metabolomics using hydrophilic interaction liquid chromatography (HILIC) and Orbitrap mass spectrometry. The method enables comprehensive analysis of biofluids for mitochondrial research.

Keywords:

Accurate mass Data-dependent acquisition HILIC High-resolution Liquid chromatography Mass spectrometry Metabolites Metabolomics Profiling Quality assurance Quality control

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A Strategy for Sensitive, Large Scale Quantitative Metabolomics

A Strategy for Sensitive, Large Scale Quantitative Metabolomics

Published on: May 27, 2014

Area of Science:

Biochemistry
Analytical Chemistry
Mitochondrial Biology

Background:

Untargeted metabolomics is crucial for profiling diverse molecules in research.
Liquid chromatography/mass spectrometry is commonly used for its broad measurement capabilities.
Hydrophilic interaction liquid chromatography (HILIC) is vital for analyzing energy pathways in mitochondrial metabolism.

Purpose of the Study:

To present a detailed protocol for untargeted metabolomics.
To enable analysis of biofluids like plasma, urine, and cerebrospinal fluid.
To facilitate the study of mitochondrial metabolism.

Main Methods:

Utilizing hydrophilic interaction liquid chromatography (HILIC) for separation.
Employing an Orbitrap high-resolution accurate mass spectrometer for analysis.
Developing a comprehensive workflow from solvent preparation to data processing parameters.

Main Results:

A robust protocol for untargeted metabolomics on biofluids.
Detailed steps for HILIC separation and Orbitrap mass spectrometry analysis.
A method to handle large, complex datasets from metabolomic profiling.

Conclusions:

The presented protocol provides a standardized approach for untargeted metabolomics.
This method enhances the analysis of mitochondrial function through biofluid profiling.
The protocol addresses the challenges of manual data analysis in complex metabolomic studies.