AcquireX Intelligent Data Acquisition Workflow

Choose from a range of pre-defined, single-click templates arranged by application and specific to each MS, or rapidly customize your own with drag-and-drop capabilities.

Regardless of how you choose to acquire your data, the method set-up is quick and simple, even with advanced workflows for profiling, identifying and characterizing your samples.

The AcquireX data acquisition workflow enhances intelligence-driven mass spectrometry to better determine in real-time which precursors to select for enhanced MS/MS or MSⁿ (MS², MS³, and/or MS⁴) interrogation (depending upon which mass spectrometer is being used), how to fragment the precursor to generate structurally diagnostic product ions, and how to connect one experiment to another for effective study evaluation.

With four data acquisition workflows, experimental flexibility to match study objectives ensures successful non-targeted small-molecule sample MSⁿ analyses. Depending upon the specific workflow being selected (see the below table), the AcquireX data acquisition workflow can automate:

• Acquisition of full scan, high-resolution, accurate-mass (HRAM) MS data to create inclusion and/or exclusion lists
• Intelligent precursor selection for tandem MS acquisition based upon exclusion and/or inclusion lists
• For iterative workflows, the dynamic management of exclusion lists

AcquireX Data Acquisition Workflow

Bypass the unwanted data acquisition of background matrix ions to preferentially drive acquisition of sample-specific precursors in one sample, or across a batch of samples, using the AcquireX background exclusion workflow. Incorporating comprehensive background exclusion increases the probability of low-level precursor selection within the chromatographic peak width. DDA methods are based on acquiring HRAM MS and tandem MS data within a user-defined cycle time; this cycle-time is based on the chromatographic peak width to ensure enough DDA cycles are acquired for accurate profiling, therefore balancing the amount of time dedicated for HRAM MS acquisition and that of the subsequent MSⁿ spectra preferentially acquired.

Like the AcquireX background exclusion workflow, the background exclusion and component inclusion workflow creates an exclusion list based upon the HRAM MS data acquisition of a representative blank or matrix sample, but also creates an inclusion list for each sample or a representative pooled sample.

The background extraction and component inclusion workflow leverages enhanced intelligence, enabling the selection of preferred ions (for example [M+H]+ adducts) from which to acquire tandem MS data. In real-time, the MS analysis can identify both [M+H]+ and [M+Na]+ ions and bypass adduct fragmentation regardless of measured ion intensity; this identification and grouping of precursors belonging to a specific compound further increases the intelligent selectivity and experimental efficiency, increasing the likelihood of increased sample coverage for compounds with high-quality fragmentation spectra.

As for the background exclusion workflow, the iterative precursor workflow creates an exclusion list based upon the HRAM MS analysis of a representative blank sample. AcquireX-DDA analysis of the sample is performed, with the exclusion list being automatically updated based upon the precursor ions being selected; additional iterative sample analyses can be performed with the exclusion list being continually updated and the process repeated until a user-defined number of replicate injections have been performed, or no more precursor ions exist for analysis.

The deep scan workflow manages replicate sample analysis and automated creation of exclusion and inclusion lists with dynamic modification of these inclusion and exclusion lists in between each replicate injection. The deep scan AcquireX workflow also has the capability to compare the lists in real-time and handle overlapping precursor m/z values to determine if it should be considered for tandem MS acquisition based on the relative ion intensity difference between the matrix blank and the sample; if the precursor ion intensity is greater in the sample by a user-defined parameter, acquisition of the tandem MS is performed.

Now that you have acquired comprehensive MSⁿ data for all your compounds, you need to identify them.

Combining the world’s largest mass spectral fragmentation library, mzCloud, with other online databases and the unknown identification capabilities of the mzLogic data analysis algorithm, means that you can be confident in assigning structures to your unknowns.

With both Compound Discoverer software and Thermo Scientific Mass Frontier software, you make the most of your data and automate the identification of your unknowns, even when there is no spectral match.

Whilst the acquisition of MS/MS data is usually sufficient for the confident structural assignment of many compounds when using mass spectral libraries, such as mzCloud, there can be certain studies or classes of compounds where additional levels of MSⁿ fragmentation can be critical in differentiating between structural isomers for example.

In the example below, the MS/MS of two structural isomer flavonoids is identical, meaning that accurate structural identification is impossible. However, the AcquireX data acquisition workflow automatically triggers neutral loss MS³ results in two different fragmentation spectra, allowing for the correct distinction between the two isomers.

For Mass Frontier software upgrades:

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