Mass Spectrometer Collision Energy Control via Precursor Ion Metrics

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Solution Overview

Problem

Current mass spectrometry techniques face challenges in optimally fragmenting precursor ions during tandem mass spectrometry of complex protein mixtures, as the optimal collision energy varies significantly with molecular weight and charge state, leading to sub-optimal or non-acceptable fragmentation, especially in automated analysis where diverse analytes are present.

Innovation Solution

The introduction of relative precursor decay (Dp) and spectral Entropy (D E ) metrics to determine collision energy based solely on molecular weight and charge state, allowing for real-time adjustment of collision energy to achieve a desired extent of dissociation, thereby eliminating the need for manual tuning and optimizing collision energy settings across a wide range of analyte characteristics.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If a fixed collision energy setting is used for all precursor ions, then the device complexity is reduced and operation is simplified, but the fragmentation quality becomes sub-optimal for analytes with different molecular weights and charge states

Engineering Contradiction:
Improveoperation simplicityVSAvoidfragmentation quality
Core Design Contradiction:
Ease of operationVSManufacturing precision

Solution Approach 1:

The system dynamically adjusts collision energy based on the molecular weight and charge state of each precursor ion. The collision energy is calculated using the formula CE = k × (MW^α) × (z^β), where k, α, and β are instrument-specific parameters. This dynamic adjustment ensures optimal fragmentation for each analyte while maintaining automated operation without manual tuning.

Inventive Principle:
Principle #15Dynamics

Solution Approach 2:

The invention changes the collision energy parameter according to the molecular weight and charge state of the precursor ion. By implementing a collision energy calculation that incorporates these variables, the system adapts the fragmentation conditions to match the specific characteristics of each analyte, thereby improving fragmentation quality across diverse protein mixtures.

Inventive Principle:
Principle #35Parameter changes

2Manufacturing precision

If manual tuning of collision energy is performed for each analyte, then the fragmentation quality is optimized, but the analysis time increases and productivity decreases

Engineering Contradiction:
Improvefragmentation qualityVSAvoidanalysis throughput
Core Design Contradiction:
Manufacturing precisionVSProductivity

Solution Approach 1:

The system performs self-service by automatically calculating the optimal collision energy for each precursor ion based on its molecular weight and charge state. The automated calculation using the formula CE = k × (MW^α) × (z^β) eliminates the need for manual tuning, thereby maintaining high fragmentation quality while preserving analysis throughput and productivity.

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The collision energy is determined in advance based on the molecular weight and charge state of the precursor ion before the fragmentation step. This preliminary calculation allows the system to prepare the optimal fragmentation conditions ahead of time, ensuring high-quality fragmentation without adding analysis time during the actual measurement process.

Inventive Principle:
Principle #10Preliminary action

3Quantity of substance

If high collision energy is applied to all precursor ions, then dissociation is enhanced, but excessive fragmentation occurs leading to loss of structural information

Engineering Contradiction:
Improvefragmentation extentVSAvoidstructural information loss
Core Design Contradiction:
Quantity of substanceVSLoss of information

Solution Approach 1:

The invention precisely controls the collision energy parameter based on the molecular weight and charge state of each precursor ion. By using the calculated collision energy CE = k × (MW^α) × (z^β), the system achieves adequate dissociation while avoiding excessive fragmentation that would lead to loss of structural information. This precise parameter control ensures that fragmentation remains within the optimal range for structural analysis.

Inventive Principle:
Principle #35Parameter changes

4Loss of information

If low collision energy is applied to all precursor ions, then structural information is preserved, but insufficient fragmentation occurs reducing analysis quality

Engineering Contradiction:
Improvestructural information retentionVSAvoidfragmentation quality
Core Design Contradiction:
Loss of informationVSManufacturing precision

Solution Approach 1:

The system adjusts the collision energy parameter upward when needed, based on the molecular weight and charge state of the precursor ion. The calculated collision energy ensures that sufficient fragmentation occurs to provide high-quality structural information, while the precise control based on analyte characteristics prevents excessive fragmentation that would lead to information loss.

Inventive Principle:
Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach enables precise control over the extent of dissociation, improving the quality of structural information obtained from mass spectra and enhancing protein identification capabilities in complex protein mixtures by ensuring optimal fragmentation across varying analyte characteristics.

Implementation Method 1

A sample or sample fraction comprising multiple proteins and/or polypeptides is input into a mass spectrometer and ionized

Methodology Applied
Scientific EffectIonization: Ionisation

Implementation Method 2

mass analyzed so as to generate a mass spectrum

Methodology Applied
Scientific EffectMass spectrometry:

Implementation Method 3

employ collision-induced dissociation to fragment precursor ions

Methodology Applied
Scientific EffectCollision-induced dissociation:

Data Source

PatentEP3631838B1Automated determination of mass spectrometer collision energy
Publication Date: 2021.09.15 THERMO FINNIGAN LLC
  • EP3631838B1 patent drawingFigure 1A~1C
  • EP3631838B1 patent drawingFigure 2~4B
  • EP3631838B1 patent drawingFigure 3A

AI summary

The present disclosure establishes new dissociation parameters that may be used to determine the collision energy (CE) needed to achieve a desired extent of dissociation for a given analyte precursor ion using collision cell type collision-induced dissociation. This selection is based solely on the analyte precursor ion's molecular weight, MW, and charge state, z. Metrics are proposed that may be used as a parameter for the "extent of dissociation", and then predictive models are developed of the CEs required to achieve a range of values for each metric. Each model is a simple smooth function of only MW and z of the precursor ion. Coupled with a real-time spectral deconvolution (m/z to mass) algorithm, methods in accordance with the invention enable control over the extent of dissociation through automated, real-time selection of collision energy in a precursor- dependent manner.