Lecture
Expanding metabolome coverage in complex matrices on the Orbitrap Excedion Pro
- at -
- ICM Saal 2
- Type: Lecture
Lecture description
Untargeted metabolomics in complex matrices is often limited by (i) insufficient sensitivity and intra-scan dynamic range that masks low-abundance metabolites in the presence of highly abundant background, and (ii) unintended in-source fragmentation that inflates feature lists and complicates annotation. Thermo Fisher Scientific™ Orbitrap™ Excedion™ Pro mass spectrometer offers new hardware and software developments designed to tackle these limitations as well as an end-to-end LC–HRAMMS workflow for expanding metabolome coverage and increasing annotation confidence.
The platform incorporates an electrodynamic ion funnel engineered for high ion transmission under mild transfer conditions, enabling reduced unintended MS1 fragmentation of labile metabolites. In a representative plant extract example (phenylalanine in tea), the fragment-to-precursor ratio decreased from 51% to 35% when comparing an Orbitrap Exploris 480 MS to the Orbitrap Excedion Pro, illustrating the potential to reduce in-source fragment interference and support cleaner precursorbased annotation.
Segmenting the mass range into narrower windows, known as the BoxCar or spectral stitching approach, has been shown to improve spectral signal-to-noise ratios and increase the detection quality of low-abundance molecules. For deeper coverage, we advanced these concepts by implementing intelligent MS1 multiplexing strategies with optimized injection times. This resulted in the Orbitrap enhanced Dynamic Range (eDR) scanning mode, that divides the mass range into two separate Orbitrap MS1 subscans, each encompassing alternating mass range windows.
In chamomile tea extracts, eDR increased the number of reproducibly detected features (CV ≤ 20%) and the number of ChemSpider-annotated compounds 1.8-fold. When coupled with AcquireX DeepScan data dependent acquisition, the workflow increased the number of MS2 spectra and fragment-based mzCloud annotations. Similar magnitude of benefits was observed for other biological matrices like human plasma, consistent with an expansion of detected peak-area dynamic range from ~5 to ~6 orders of magnitude.
Data processing is supported by Compound Discoverer 3.5 with automated in-source fragment grouping and mzCloud 2.0 library matching.
The platform incorporates an electrodynamic ion funnel engineered for high ion transmission under mild transfer conditions, enabling reduced unintended MS1 fragmentation of labile metabolites. In a representative plant extract example (phenylalanine in tea), the fragment-to-precursor ratio decreased from 51% to 35% when comparing an Orbitrap Exploris 480 MS to the Orbitrap Excedion Pro, illustrating the potential to reduce in-source fragment interference and support cleaner precursorbased annotation.
Segmenting the mass range into narrower windows, known as the BoxCar or spectral stitching approach, has been shown to improve spectral signal-to-noise ratios and increase the detection quality of low-abundance molecules. For deeper coverage, we advanced these concepts by implementing intelligent MS1 multiplexing strategies with optimized injection times. This resulted in the Orbitrap enhanced Dynamic Range (eDR) scanning mode, that divides the mass range into two separate Orbitrap MS1 subscans, each encompassing alternating mass range windows.
In chamomile tea extracts, eDR increased the number of reproducibly detected features (CV ≤ 20%) and the number of ChemSpider-annotated compounds 1.8-fold. When coupled with AcquireX DeepScan data dependent acquisition, the workflow increased the number of MS2 spectra and fragment-based mzCloud annotations. Similar magnitude of benefits was observed for other biological matrices like human plasma, consistent with an expansion of detected peak-area dynamic range from ~5 to ~6 orders of magnitude.
Data processing is supported by Compound Discoverer 3.5 with automated in-source fragment grouping and mzCloud 2.0 library matching.
