Evaluation of a Prototype Orbitrap Astral Zoom Mass Spectrometer for Quantitative Proteomics─Beyond Identification...

Area of Science:

• Proteomics
• Analytical Chemistry
• Mass Spectrometry

Background:

• Quantifying advances in mass spectrometry (MS) instrumentation beyond standard peptide and protein detection is challenging.
• Evaluating new MS platforms requires robust performance metrics for cross-platform comparisons.

Purpose of the Study:

• To evaluate a modified Orbitrap Astral Zoom mass spectrometer (MS) prototype.
• To compare its performance against the standard Orbitrap Astral MS.
• To establish a generalizable framework for evaluating MS performance based on ion detection.

Main Methods:

• Implemented an ion calibration framework to convert signal intensity to ions per second.
• Compared precursor and protein identifications, ion beam utilization, and quantitative precision between the prototype and standard MS.
• Introduced new metrics in Skyline for reporting ion counts in spectra.

Main Results:

• The Orbitrap Astral Zoom prototype outperformed the standard Orbitrap Astral MS in precursor and protein identifications, ion beam utilization, and quantitative precision.
• The prototype sampled 23.1% more ions per peptide than the standard MS.
• Increased ion beam utilization led to improved sensitivity and quantitative precision.

Conclusions:

• The Orbitrap Astral Zoom prototype is a high-performance platform for data-independent acquisition proteomics.
• A novel framework for MS performance evaluation based on ion detection was established.
• The findings provide a generalizable method for assessing MS performance.