Signal Processing Workflow for Suspect Screening in LC × LC-HRMS: Efficient Extraction of Pure Mass Spectra for
Paul-Albert Schneide1,2, Oskar Munk Kronik3
1Department of Food Science, University of Copenhagen, 1958 Frederiksberg, Denmark.
Analytical Chemistry
|January 8, 2025
Summary
A new MF + MCR workflow enhances data processing for comprehensive two-dimensional liquid chromatography-high-resolution mass spectrometry (LC × LC-HRMS) in data-independent acquisition (DIA). This method improves compound identification, especially for trace analytes.
Area of Science:
- Analytical Chemistry
- Chromatography
- Mass Spectrometry
Background:
- Data processing workflows for LC × LC-HRMS in DIA are limited.
- Extracting pure mass spectra is crucial for reliable compound identification.
- Existing methods struggle with complex matrices and trace analytes.
Purpose of the Study:
- To develop and evaluate a novel two-step workflow (MF + MCR) for extracting pure mass spectra from LC × LC-HRMS DIA data.
- To benchmark the proposed workflow against existing methods: peak apex (PAM), mass filtering (MF) alone, and multivariate curve resolution (MCR) alone.
- To assess the workflow's effectiveness in identifying suspect compounds and extracting purer mass spectra, particularly for trace-level analysis.
Main Methods:
- A two-step data processing workflow termed MF + MCR was developed.
- Mass filtering (MF) algorithm groups ions based on elution profile similarity across both chromatographic dimensions.
- Multivariate curve resolution (MCR) deconvolutes filtered data to address coelution.
Main Results:
- The MF + MCR workflow identified 25 suspect compounds, outperforming MF (23), MCR (16), and PAM (10).
- MF + MCR extracted statistically significantly purer mass spectra compared to PAM (p=0.003) and MCR (p=0.04).
- Adequate preprocessing via MF is essential for MCR's effectiveness in trace-level analysis.
Conclusions:
- The MF + MCR workflow effectively extracts pure mass spectra from LC × LC-HRMS DIA data.
- This approach enables more reliable compound identification, especially for trace analytes in complex samples like wastewater effluent.
- Utilizing elution profiles in both dimensions is key to improving spectral purity and analytical performance.
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