Validation of the Mass-Extraction-Window for Quantitative Methods Using Liquid Chromatography High Resolution Mass...

Area of Science:

• Analytical Chemistry
• Mass Spectrometry
• Chromatography

Background:

• Recent high-resolution mass spectrometers (HRMS) are shifting quantitative liquid chromatography-mass spectrometry (LC-MS) analysis.
• HRMS in full scan mode allows sensitive quantification and qualitative analysis, enabling retrospective investigations.
• A need exists for HRMS-specific guidance in validated quantitative analyses to avoid false positives/negatives.

Purpose of the Study:

• To investigate key parameters for validating quantitative LC-HRMS analyses: mass accuracy (MA) and mass-extraction-window (MEW).
• To propose a procedure for calculating rational MEW width for validated quantitative LC-HRMS methods.

Main Methods:

• Investigated MA and MEW parameters crucial for quantitative LC-HRMS validation.
• Performed MA measurements across four different LC-HRMS platforms.
• Developed MA-parameters, graphs, and equations to determine MEW width.

Main Results:

• Experimentally determined MEW values varied from 5.6 to 16.5 ppm.
• MEW values were influenced by the HRMS platform, working environment, calibration, and analyte.
• The proposed procedure ensures fit-for-purpose MEW determination.

Conclusions:

• The proposed method provides a validated approach for determining MEW in quantitative LC-HRMS.
• This procedure helps prevent false detections in quantitative LC-HRMS analyses.
• This work addresses a critical need for HRMS-specific validation guidance.