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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
Published on: May 20, 2013
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The SIMPLIFY Protocol: A Monophasic Extraction System Suitable for Exposomics, Metabolomics, Lipidomics, and
Anh Hoang Nguyen1, Victor Castro-Alves1, Emilia Holmström Flores1
1Man-Technology-Environment (MTM) Research Centre, School of Science and Technology, Örebro University, Örebro SE-701 82, Sweden.
Analytical Chemistry
|November 19, 2025
Summary
The SIMPLIFY Protocol offers a unified method for simultaneous extraction of exogenous and endogenous molecules, advancing exposomics and metabolomics research. This efficient approach supports high-throughput clinical studies by simplifying workflows and reducing resource consumption.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Environmental Health
Background:
- Understanding human health and disease necessitates analyzing complex interactions between metabolism and environmental factors.
- Current clinical research faces challenges in acquiring multidimensional biochemical data due to resource, time, and budget constraints.
Purpose of the Study:
- Introduce the SIMPLIFY Protocol, a novel unified monophasic extraction method.
- Enable simultaneous extraction of both exogenous and endogenous molecules for comprehensive analysis.
Main Methods:
- Evaluated the SIMPLIFY Protocol against established methods like methanol (MeOH) and acetonitrile (ACN) protein precipitation, and the Folch method.
- Utilized certified reference materials and diverse sample types for validation.
- Assessed analyte abundance and reproducibility across different sample matrices.
Main Results:
- The SIMPLIFY Protocol demonstrated comparable performance to in-house extraction techniques.
- The method showed enhanced versatility for applications including derivatization and proteomics.
- Successfully profiled approximately 800 compounds, including polar, semipolar, and nonpolar molecules, with minor limitations for triacylglycerols.
Conclusions:
- The SIMPLIFY Protocol provides a versatile and efficient solution for simultaneous multi-class metabolite and xenobiotic extraction.
- Its streamlined, automatable design is ideal for high-throughput exposomics, metabolomics, and lipidomics in large-scale clinical studies.
- This method addresses critical needs for efficiency, cost-effectiveness, and sample preservation in clinical research.
