On the first day of June 2025, at the ASMS 2025 conference held in Baltimore, Thermo Fisher Scientific unveiled its new product line for the year, featuring Thermo Scientific ™ Orbitrap ™ Excedion ™ The Pro, named after Thermo Fisher's next-generation 2-in-1 ultra-high resolution mass spectrometer, has become the successor to Thermo Fisher's glorious production line over the past fifteen years. It is the star product of the third generation "Ex" family after the Q Exactive (QE series) and Orbitrap Explores (OE series). We hope that it can inherit the "workhorse" family style of the "Ex" family and work tirelessly to bring users an extraordinary experience of "Exceed".

So in this article, let's follow the Baltimore wind and experience the brand new Thermo Scientific ™ Orbitrap ™ Excedion ™ Pro it~

At the front end of the instrument, in order to achieve higher transmission efficiency and better performance for unstable compounds, Orbitrap ™ Excedion ™ The front end of Pro has replaced the OptaMax Plus ion source, which is the high-end product Stellar from our new generation Thermo Fisher quantitative product line released last year ™ The ion source released together with the mass spectrometer (also makes us eagerly look forward to Orbitrap) ™ Excedion ™ Pro's future performance in the quantitative field), it can provide better spray stability in a higher temperature range. At the same time, it is equipped with a new generation of electric ion funnels similar to Orbitrap Ascend, designed specifically for unstable compounds or modified peptide segments that are prone to change, providing gentler front-end transport and reducing in source losses. The combination of the two swords complements each other perfectly.

In addition, for proteomics users, we will also release a completely new nanoliter ion source OptiSpray this year, bringing users better performance while providing a simpler, more convenient, and stable user experience. And the new nano source region can directly support multiple modes such as calibration and direct injection.

Orbitrap ™ Excedion ™ Pro brings innovative eDR (enhanced dynamic range) technology. Simply put, we split the original Full MS scan into two sub scans, allowing for better ion transport and accumulation, and achieving an effect equivalent to gas phase horizontal separation. Finally, the two sub scans will be automatically concatenated and output through software. EDR technology will significantly improve the signal-to-noise ratio of low abundance parent ions, bring better result confidence, and enhance the performance of LOQs/LDs instruments. The signal amplification effect of such a first-order spectrum also enhances the sensitivity of the primary parent ions in proteomics and improves the interference caused by high abundance proteins on identification.

In order to further improve the qualitative and quantitative performance of proteomics, we hope to provide users with the ability to quantify target peptides simultaneously without significantly affecting the DIA acquisition speed. We named it Hybrid DIA technology and combined it with Adaptive RT technology. As shown in the experiment below, within a 30 minute gradient time, 450 target peptides can be quantified without sacrificing DIA performance. The linear range of peptide quantification is 62.5 amol to 1000 fmol.

In the new Orbitrap ™ Excedion ™ On Pro, our Orbitrap detector has undergone further evolution, coupled with Bent Trap pre accumulation technology, providing longer effective ion implantation time. At the same time, the scanning speed of Orbitrap reached 70Hz, which enabled Orbitrap to ™ Excedion ™ Pro has shown better performance in short gradients and small injection volumes. As shown in the figure, on a 9-minute ultra short gradient, Bent Trap pre accumulation technology can bring corresponding improvements from 6ng to 200ng injection volume, and the improvement is more significant at lower sample volumes.

In addition, for the TMT-32plex and TMT-35plex samples, the new Orbitrap ™ Excedion ™ The Phi SDM algorithm on Pro will also bring TurboTMT 45k and TurboTMT 60k algorithms to ensure users can quantitatively report the baseline separation of ions.

Different fragmentation methods play a crucial role in practical applications. In the traditional first two generations of instruments, the 2-in-1 product line only provided HCD fragmentation (as well as in source CID) mode, while in the new Orbitrap ™ Excedion ™ On Pro, we will provide the fragmentation method of EASY Electron Transfer Dissection (ETD) and also enable EThcD mode. Simply put, in the all-new Orbitrap ™ Excedion ™ On Pro, the parent ion is captured in IRM and reacts with fluoranthene reagent here, producing ETD fragments that are returned to C-Trap. At this point, we can choose to continue accelerating into IRM for EThcD or directly enter Orbitrap for detection.

In practical applications, the fields of biopharmaceuticals and proteomics, ETD and EThcD modes have a wide range of application scenarios. When we need to perform more precise characterization of protein sequences or peptide modifications, we have a more rigorous requirement for sequence coverage and confirmation information of modification sites. In this case, ETD and EThcD patterns will become our powerful helpers.

Firstly, when faced with the identification of modifications such as intact glycopeptides, the fragmentation mode of EThcD can break peptide bonds while preserving the connection between the sugar chain and amino acids, providing us with both modification site information and peptide sequence information.

Another case comes from top-down or middle-level experiments. Compared to a single HCD mode, multiple fragmentation modes such as ETD and EThcD addition can provide us with higher sequence coverage, which is beneficial for our fine characterization of protein sequences. As shown in the figure below, especially in the EThcD mode, we can achieve higher sequence coverage than the pure ETD mode (53% vs 40%). Meanwhile, the all-new Orbitrap ™ Excedion ™ The mass range of Pro can be extended to m/z 12000, which benefits both complete protein and top-down detection.

In terms of coverage, in addition to protein sequence coverage experiments such as Top down or Middle down, peptide level identification such as immunopeptidomics can also benefit.

In addition to assisting with sequence coverage, in experiments such as denovo or peptide mapping, we often need to confirm certain amino acid sequences, such as the resolution of I and L and the resolution of D and isoD, which can be confirmed through the fragmentation mode of ETD.

Overall, Thermo Fisher's new Orbitrap this year ™ Excedion ™ The Pro mass spectrometer brings us faster scanning speed, better sensitivity, new eDR and Hybrid DIA technologies, improvements in ETD, EThcD fragmentation modes, and m/z range expansion. We look forward to the new Orbitrap ™ Excedion ™ The Pro mass spectrometer can bring us more efficient, fast, and accurate protein qualitative and quantitative analysis, as well as better experimental results for biopharmaceutical characterization, top-down and other macromolecular experiments.