Orbitrap Exploris 480

Hybrid quadrupole-Orbitrap mass spectrometer

The Thermo Scientific Orbitrap Exploris 480 is an advanced, intelligence-driven instrument, built for ultimate performance and ease of use. The hardware has been designed to address laboratory requirements for maximum uptime and easy serviceability. The instrument control software is shared with nextgeneration Thermo Scientific™ Tribrid™ and Thermo Scientific™ TSQ triple quadrupole mass spectrometers for simplicity of operation, reduced training needs, and easy transfer of methods and results from one research stage to the next. The Orbitrap Exploris 480 mass spectrometer has unique capabilities which impart extra speed, depth, and certainty to enable you to run more experiments or to gain deeper insights into your research.

Visit the Planet Orbitrap website for more information

12/1/2020 Exploris480 system 1 (SN MA10302C, UW tag 2033415) Installed
3/23/2023 Exploris480 system 2 (SN MA10852C, UW tag 2033420) Installed

Specifications

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    Scan Rate
  • Up to 40 Hz at resolution setting 7500 at m/z 200
  • Multiplexity: up to 20 precursors/scan
    Resolution
  • up to 480 000 at m/z 200
    Mass accuracy
  • < 3 ppm RMS drift over 24 hours using external calibration
  • < 1 ppm RMS drift over 24 hours using internal calibration
    Sensitivity
  • Dynamic range > 5000 within a single Orbitrap mass analyzer spectrum
  • MS/MS: 50 fg reserpine on column S/N 100:1
  • SIM: 50 fg reserpine on column S/N 150:1
    Features
  • Higher Energy Collisional Dissociation (HCD)
  • EASY-IC™ (internal calibration) source
  • Quadrupole mass filter
  • Ion Routing Multipole (IRM) trapping, focuses and routes ions more effectively
  • Orbitrap mass analyzer
  • Broader range of experiments with multiple scan modes: data-dependent acquisition (DDA), dataindependent acquisition (DIA), spectral multiplexing (MSX), TopN and new TopSpeed (TopS), BoxCar DIA and DDA experiments, tSIM, tSIM MSX, ddMS/MS, targeted MS/MS

Instrument control software

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Instrument calibrations

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Resolving Power and Transient Length

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Here is a full table of the available resolution settings. Note: higher resolution does not always result in better mass accuracy. The higher the resolution increases your ability to distinguish between m/z ions. As you can see in this table, there is always a trade of higher resolution with time. The Exploris480 allows acquisition of transients at resolution settings ranging from 7500 to 480,000 at m/z 200 (16 and 1024 ms transients, respectively).
To fully take advantage of the parallel fill and detect capabilities of the Exploris480, we need to balance the max fill times with the transient length. For resolving power 15,000, detection time is about 32 ms. There is always a small amount of inter-scan delay. Up to 22 ms can be used to fill the C-trap with ions- and without increasing cycle time.

Res. at m/z 200 Transient length [ms]Approx. scan speed [Hz]"Free" fill time [ms]
7500 16 40
15,000 32 22 22
30,000 64 12 54
45,000 10
60,000 128 7 118
120,000 256 3 246
180,000 2
240,000 512 1.5 502
480,000 1024 0.7 1014

TMT instrument Orbitrap Exploris480 (Tune1.1)

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The table below lists instrument method settings recomended by Thermo for the Lumos with Tune 3.3.
with APD (Advanced Peak Detection) on
based on this document: TMT/TMTpro Instrument Acquisition Parameter Settings

Properties MS2 120 min TMT 11plex MS2 120 min TMT 16plex
Resolution Full MS 120,000 120,000
AGC target Full MS 100% (4e5) 100% (4e5)
MS max IT, ms 50 50
Scan range, m/z 400 - 1400 400 - 1400
Top Speed, s 3 3
MS2 max IT, ms 105 120
MS2 Isolation window, Th 0.7(2-3)-0.5(4+) 0.7
MS2 NCE, % 38-40 32
MS2 Intensity threashold 5e4 5e4
Dynamic exclusion, s 60, single charge 60, single charge
MS2 Resolution 50,000 50,000
MS2 AGC target 200% ( 1e5 ) 200% ( 1e5 )
First mass m/z 110 m/z 110

HPLC setup

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the following documents are guidelines to setup the capillary trap/column on our LC-MS systems, this is intended as a supplement to the hands on training required to use the UWPR instruments.

Instrument Settings

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The table below lists some of the instrument method settings we tested on our Exploris480.
We use a EASYnLC UPLC with home made columns (35cm x 75μm, Reprosil Pur C18AQ 120Å 5μm) and trap column (3cm x 100μm, Reprosil Purc C18AQ 120Å 5μm), with 6-36% B (80% ACN, 0.1%FA) in 90min gradient. The sample is a Hela cell whole cell tryptic digest (ThermoFisher), 100 ng on column injection. We typically get ~22000 unique peptide ID's and 30000 PSM's with Comet search and peptide prophet p>0.9,
The results below are based on the number of unique peptide ID's with Comet search and peptide prophet p>0.9.

Tested SettingBest Result (100%)
Software Version Exploris 4.2 SP1
MS2 res unique ID's MS2 7.5k (89%), 11.5k (96%), 15k (98%), 22.5k (100%), 30k (89%), 45k (71%)
MS2 res PSM's MS2 7.5k (88%), 11.5k (95%), 15k (98%), 22.5k (100%), 30k (89%), 45k (69%)
. .
Software Version Exploris 4.0
10 plex TMT unique ID's MS2 res 45k (100%), turbo 15k (60%), turbo 30k (98%)
10 plex TMT PSM's MS2 res 45k (99%), turbo 15k (64%), turbo 30k (100%)
% PSM's with signal in all TMT channels 45k (99.5%), turbo 15k (92.9%), turbo 30k (99.6%)
MS2 res unique ID's MS2 7.5k (89%), 15k (100%), 30k (92%), 45k (72%), 60k (59%)
MS2 res PSM's MS2 7.5k (87%), 15k (100%), 30k (92%), 45k (70%), 60k (57%)
. .
Software Version Exploris 2.0
S-lens setting, unique ID's 30 (93%), 40 (97%), 50(100%), 60 (92%)
S-lens setting, PSM's 30 (87%), 40 (94%), 50(100%), 60 (91%)
Gradient type, unique ID's 3-19-29%B (100%), 6-35%B (99.9%)
Gradient type, PSM's 3-19-29%B (94%), 6-35%B (100%)