By upgrading your SYNAPT^TM MS series instrument to full HDMS^TM capability you can take advantage of high efficiency ion mobility separations (IMS) to extend your research to a new dimension. The HDMS upgrade will enable you to differentiate samples based on size, shape, and charge, as well as mass. This provides a range of new or improved analytical benefits such as conformational analysis, the ability to separate different compound classes, remove background and interferences, or carry out comprehensive structural characterization.

Ion mobility/HDMS mode expands your capabilities beyond those of your present SYNAPT MS series instrument by incorporating:

• Triwave^TM Technology: Provides the ability to perform ion mobility separations at the limits of MS detection and also combine with a dual collision cell arrangement for a range of analytical benefits.
  • IMS - Separate isobaric components, derive molecular conformation values (collision cross section), remove background and reduce complexity (increase information content by separating and removing interferences), and separate different compound classes.
  • CID/IMS and IMS/CID - Reduce complexity of MS/MS spectra for complex analytes (e.g. in top-down protein sequencing) or produce independent MS/MS spectra of separated or partially (IMS) separated species for more conclusive structural elucidation.
  • Time aligned parallel (TAP) fragmentation (CID/IMS/CID): Provides comprehensive structural (MS/MS/MS) information in a single experiment for characterization studies.
• High duty cycle (HDC) mode: Enables significant sensitivity enhancements over a wide m/z range
• DriftScope^TM Software mobility environment: For powerful and intuitive interrogation of 3D and 4D datasets that provide greater levels of productivity for users of all experience levels. The software incorporates a range of features for the enhanced interactive visualization, manipulation, interpretation and exporting of multidimensional data.

MALDI source users can also benefit significantly from the HDMS upgrade particularly with imaging applications using the Waters High Definition Imaging (HDI^TM) MALDI solution. HDI MALDI helps generate images without interference from background ions of similar mass, leading to more precise localization of the drug or metabolite of interest.

Triwave Technology

Triwave™ Technology is the enabling system behind the SYNAPT™ MS series HDMS™ field upgrade. It uses three T-Wave™* ion guides that trap ions, separate them by their ion mobility and transfer them to the orthogonal acceleration time-of-flight analyzer for high-resolution mass analysis. The trap and transfer regions can also be used for fragmentation analyses.

The Triwave Technology’s innovative configuration overcomes the major performance drawbacks of conventional ion mobility spectrometry (IMS) by ensuring that IMS does not compromise the base sensitivity of the instrument.

*The Traveling wave device described here is similar to that described by Kirchner in US Patent 5,206,506; 1993.

The combination of MALDI and the ion mobility field upgrade allows MALDI users to isolate analyte ions from multiply charged background ions & matrix ions, enabling the separation of molecular species by structural class. This is particularly important for MALDI imaging applications as ion mobility separation has been shown to separate isobaric interferences from imaging data obtained from highly complex biological samples, so increasing confidence in the resulting image.

This example illustrates how singly charged ions can be separated from multiply charged components using MassLynx™ or DriftScope™ Software, thereby simplifying data interpretation. Further investigation, (shown in the detailed image), reveals the separation of isobaric peptides by drift time. A subsequent database search confirms the presence of isobaric peptides, resulting in increased protein coverage for the experiment.

DriftScope Sofware shows data of different charge states separated by drift time.  Detail shows isobaric precursors separated by drift time.

Unlock the full power of the SYNAPT MS series HDMS field upgrade

DriftScope^TM Mobility Environment Software incorporates new technologies to enable comprehensive retrieval of information contained within HDMS^TM (ion mobility) datasets while enabling easier data interpretation (for users of all experience levels) through a series of data visualization and data export enhancements:

Powerful and automated peak detection (novel APEX 3D and 4D technology)

• Comprehensively reveals all components within any HDMS data (e.g. IMS/MS, LC/IMS/MS and TAP fragmentation), and increases apparent specificity of each analysis
• Eliminates subjectivity from data interpretation, providing highly reproducible results across analyses and users

Visualization enhancements (e.g. 3D peak profile display, x/y axis switching, peak/raw data overlay, lock-axis function, ability to turn raw data on/off)

• Enables easier and more intuitive data interpretation
• Provides convenient comparison of peaks and supporting raw data

Data export functionality (raw data or peak list export and peak copy/paste function)

• Facilitates data transfer to downstream processing routines
• Streamlines data processing workflows

Enhanced analytical power

• Confident detection of partially (IM) separated compounds (e.g. Isomers or Conformers)
• Visualizing new molecular trends in complex mixtures (e.g. Crude Oil, PEG, MALDI Imaging)
• More efficient characterization with IMS/CID and TAP fragmentation (export of detected peaks directly to Massfragment software)

DriftScope mobility environment software provides automated peak detection from 2D (Chromatograms and Mass Spectra), or 3D graphical plots. The axes are interchangeable, each pixel representing an ion, and its color, the intensity. The peaks are represented as colored (green) dots and sticks. The data shown is from the Electrospray (infusion-based) IMS-MS analysis of a Polyethylene Glycol (PEG) sample.

Conformation (IMS) Studies

Measuring the mobility (or drift time) of an ion can yield information about its structure, which you can then use to investigate the size and shape of small and large molecules.

Molecular conformation is critical to the integrity of many biological systems, which means that increased knowledge of the shapes of biologically relevant components (such as proteins, peptides, carbohydrates, and nucleotides) can benefit many research areas.

Analysis of the native and reduced forms of the intact protein Hen Egg Lysosyme using Mobility-TOF (HDMS™) mode on the SYNAPT™ HDMS System. The DriftScope™ Software plot shows m/z versus drift time. Upon acidification and reduction of the native form (single drift time distributions), multiple drift time distributions are observed for the two charge-states shown. This indicates a change in the overall size and shape (topology) of the protein has taken place. Average rotational cross-sections can be determined for the individual conformers from these data.

Reduce Sample Complexity

The SYNAPT™ MS series instrument HDMS™ upgrade provides a unique way to increase the information content for a variety of applications. DriftScope™ Software eliminates common interferences from an analysis, providing access to lower-level components:

• Separation and removal of interferences; PEG contaminants, chemical noise etc.
• Enhanced detection limits; charge state separation of peptides, PEG, glycans etc.
• Increased information content; structural characterization e.g. Top-Down Proteomics

UPLC®/IMS-TOF sample analysis of a rat bile sample containing the dosing vehicle (PEG), parent drug and its metabolites. The circled regions can be easily removed from the data set using DriftScope Software (e.g. PEG dosage vehicle). This provides a simple, unique route to enhanced MS detection of low-level components.

Analysis of released glycans from IgG using a MALDI SYNAPT HDMS System showing the effect of using ion mobility to remove background ions.

Enhanced Fragmentation

The SYNAPT™ MS series instrument HDMS™ upgrade provides an expanded range of fragmentation protocols for structural characterization studies through use of the trap and transfer areas of the Triwave™ to induce fragmentation.

Time Aligned Parallel (TAP) Fragmentation Triwave Technology allows the combination of high-efficiency ion mobility spectrometry with a dual-stage fragmentation process for detailed structural characterization.

Features

• Provides first- and second-generation product ions from a precursor ion - in one experiment
• Maximizes the information content from each sample
• Can be used in a wide range of applications, such as protein PTM, lipid, and small molecule analysis

TAP Fragmentation: All first generation product ions are fragmented in the Transfer T-Wave – a Parallel process; the second generation product ions are Time Aligned to original first generation ions.

Equipment and Installation

The SYNAPT^™ MS series HDMS^™ system upgrade is available for the following instruments: 

• SYNAPT MS, MALDI SYNAPT MS
• SYNAPT G2 MS, MALDI SYNAPT G2 MS
• SYNAPT G2-S MS, MALDI SYNAPT G2-S MS
• SYNAPT G2-Si MS, MALDI SYNAPT G2-Si MS

The upgrade kit includes:

• Provision of full SYNAPT HDMS capability (firmware and control software update)
• DriftScope^™ Mobility Environment Software
• External gas cell regulator (SYNAPT MS only)
• Replacement instrument badge and illumination pcb, where applicable

The upgrade requires installation of the above hardware and new system software and firmware to reconfigure the present Triwave gas cell technology to full High Definition MS functionality. DriftScope Mobility Environment software will also be installed. The work is carried out by a certified Waters Field Service Engineer who also provides initial operator training.

On completion of the HDMS upgrade, the total MS instrument including MALDI source where applicable, is warranted against component failure for a further 90 days.