AP-ECD Mass Spectrometry for Parent-Fragment Ion Association

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Solution Overview

Problem

Existing atmospheric pressure Electron Capture Dissociation (AP-ECD) mass spectrometers face challenges in associating parent ions with their fragment ions due to the high-pressure environment, leading to complex spectra and limited analytical utility, especially when analyzing mixtures.

Innovation Solution

The method involves performing ECD and/or ETD reactions at atmospheric pressure, using an RF ion guide or ion trap to interact electrons or reagent anions with parent ions, and continuously associating fragment ions with their parent ions through simultaneous mass analysis, leveraging liquid chromatography or ion mobility separation to correlate elution or drift times.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional ion selection techniques are used in high pressure region, then parent ions can be selected for fragmentation, but the high pressure environment prevents effective ion selection and association

Engineering Contradiction:
Improveion association accuracyVSAvoidion selection capability
Core Design Contradiction:
Measurement precisionVSEase of operation

Solution Approach 1:

The invention segments the mass spectrometer into distinct pressure regions: a high pressure ion source region for ECD/ETD fragmentation and a low pressure analysis region for ion selection and detection. This spatial segmentation allows each region to operate under optimal conditions without interfering with the other, resolving the contradiction between maintaining high pressure for fragmentation and low pressure for precise ion selection.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention introduces an intermediary transfer region with pressure reduction mechanisms (such as skimmers or capillaries) that mediates the transition of ions from the high pressure fragmentation region to the low pressure analysis region. This intermediary structure allows ions to be transferred while maintaining the pressure differential, enabling both high pressure fragmentation and low pressure ion selection to coexist.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Ease of operation

If single parent ion selection is used, then parent-ion/fragment-ion association is simple, but duty cycle is low as other parent ions are discarded

Engineering Contradiction:
Improvedata processing simplicityVSAvoidanalytical throughput
Core Design Contradiction:
Ease of operationVSProductivity

Solution Approach 1:

The invention enables continuous useful action by allowing simultaneous fragmentation and analysis of multiple parent ions in parallel. The high pressure region continuously fragments all incoming parent ions via ECD/ETD, while the low pressure region continuously analyzes and selects specific ions. This continuous parallel processing eliminates the duty cycle losses inherent in sequential single-ion selection methods.

Inventive Principle:
Principle #20Continuity of useful action

Solution Approach 2:

The invention adds the dimension of pressure as a separating variable, allowing ions to be processed in three dimensions: mass-to-charge ratio (m/z), time, and pressure. By utilizing pressure differential as an additional dimension, the system can simultaneously perform fragmentation at high pressure and selective analysis at low pressure, effectively multiplying the analytical capacity beyond what single-pressure systems can achieve.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

3Adaptability or versatility

If AP-ECD is used for mixture analysis, then comprehensive fragmentation occurs, but spectra become complex with multiple ion types making association difficult

Engineering Contradiction:
Improvemixture analysis capabilityVSAvoidspectral complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The invention extracts specific ion types from the complex mixture by utilizing mass-selected ion transmission through the pressure differential system. The low pressure region allows for selective transmission of fragment ions while excluding other ion types (such as solvent ions, dopant ions, and unreacted parent ions) through precise mass filtering and mobility separation, effectively extracting the desired fragment ion information from the complex spectral mixture.

Inventive Principle:
Principle #2Taking out (Extraction)

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 effective fragmentation and association of parent ions with their fragment ions, simplifying data processing and improving the analytical utility of AP-ECD mass spectrometry by correlating ions based on their elution or drift times, enhancing the identification of biomolecules and disulphide linked species.

Implementation Method 1

a method of mass spectrometry comprising: (i) causing a beam of electrons or reagent anions to interact with parent ions within an RF ion guide or ion trap so as to fragment the parent ions

Methodology Applied
Scientific EffectElectron Capture Dissociation:

Implementation Method 2

causing a beam of electrons or reagent anions to interact with parent ions within an RF ion guide or ion trap so as to fragment the parent ions

Methodology Applied
Scientific EffectElectron Transfer Dissociation:

Implementation Method 3

a mass analysing device arranged and adapted to mass analyse the parent ions and the fragment or product ions

Methodology Applied
Scientific EffectMass Spectrometry:

Implementation Method 4

leveraging liquid chromatography or ion mobility separation to correlate elution or drift times

Methodology Applied
Scientific EffectLiquid Chromatography: Chromatography

Implementation Method 5

leveraging liquid chromatography or ion mobility separation to correlate elution or drift times

Methodology Applied
Scientific EffectIon Mobility Separation:

Data Source

PatentEP3598478B1Method of ms/ms mass spectrometry
Publication Date: 2025.03.26 MICROMASS UK LTD
  • EP3598478B1 patent drawingFigure 1~2

AI summary

A method of mass spectrometry and a mass spectrometer performing mass analysing parent ions; subjecting parent ions to ECD and/or ETD thereby producing fragment ions and mass analysing the resulting fragment ions; subjecting parent ions to ECD and/or ETD thereby producing intermediate ions, which are non-dissociated parent ions held together by covalent interactions and/or charge reduced parent ions, and subjecting these intermediate ions to a fragmentation technique other than ETC and/or ECD thereby producing fragment ions and analysing the resulting fragment ions; and associating parent ions with fragment ions.