Orbitrap Explained

In mass spectrometry, Orbitrap is an ion trap mass analyzer consisting of an outer barrel-like electrode and a coaxial inner spindle-like electrode that traps ions in an orbital motion around the spindle.[1] [2] The image current from the trapped ions is detected and converted to a mass spectrum by first using the Fourier transform of time domain of the harmonic to create a frequency signal which is converted to mass.

History

The concept of electrostatically trapping ions in an orbit around a central spindle was developed by Kenneth Hay Kingdon in the early 1920s.[3] The Kingdon trap consists of a thin central wire and an outer cylindrical electrode. A static applied voltage results in a radial logarithmic potential between the electrodes. In 1981, Knight introduced a modified outer electrode that included an axial quadrupole term that confines the ions on the trap axis.[4] Neither the Kingdon nor the Knight configurations were reported to produce mass spectra.

In 1986, Professor Yuri Konstantinovich Golikov in USSR conceived the core principles and theory of Orbitrap analyzer, and filed for patents in USSR for Orbitrap analyzer. Golikov, leading a team at the St. Petersburg State Pedagogical University's Radiophysics Faculty, laid the theoretical groundwork for Orbitrap technology as its original inventor in certificate USSR No. 1247973 in 1986. As the original inventor of Orbitrap analyzer, Golikov remarked "Based on my ideas, analytical instruments with record parameters were built, but unfortunately not in Russia, but abroad."[5]

Contrary to popular belief, Alexander Makarov is not the original inventor of Orbitrap analyzer and he only reduced the Orbitrap analyzer to practice and further developed and commercialized the Orbitrap analyzer at Thermo Fisher Scientific. Reflecting on his early interaction with Golikov, Alexander Makarov recalled, "As a fifth-year student at MIPT, I entered one of the numerous rooms at the Polytechnic Institute, where I was met by Yuri Konstantinovich Golikov. I was holding excerpts (photocopies were not so accessible then) from the author's certificate USSR № 1247973 to which I have referred in all my works on the Orbitrap™ analyzer since then."[5] [6]

Alexander Makarov's effort in commercializing Orbitrap analyzer at the end of the 1990s started a sequence of technology improvements which resulted in the commercial introduction of this analyzer by Thermo Fisher Scientific as a part of the hybrid LTQ Orbitrap instrument in 2005.[7] [8]

Principle of operation

Trapping

In the Orbitrap, ions are trapped because their electrostatic attraction to the inner electrode is balanced by their inertia. Thus, ions cycle around the inner electrode on elliptical trajectories. In addition, the ions also move back and forth along the axis of the central electrode so that their trajectories in space resemble helices. Due to the properties of the quadro-logarithmic potential, their axial motion is harmonic, i.e. it is completely independent not only of motion around the inner electrode but also of all initial parameters of the ions except their mass-to-charge ratios m/z. Its angular frequency is: ω =, where k is the force constant of the potential, similar to the spring constant.

Injection

In order to inject ions from an external ion source, the field between the electrodes is first reduced. As ion packets are injected tangentially into the field, the electric field is increased by ramping the voltage on the inner electrode. Ions get squeezed towards the inner electrode until they reach the desired orbit inside the trap. At that moment ramping is stopped, the field becomes static, and detection can start. Each packet contains a multitude of ions of different velocities spread over a certain volume. These ions move with different rotational frequencies but with the same axial frequency. This means that ions of a specific mass-to-charge ratio spread into rings which oscillate along the inner spindle.

Proof-of-principle of the technology was carried out using the direct injection of ions from an external laser desorption and ionization ion source. This method of injection works well with pulsed sources such as MALDI but cannot be interfaced to continuous ion sources like electrospray.

All commercial Orbitrap mass spectrometers utilize a curved linear trap for ion injection (C-trap). By rapidly ramping down trapping RF voltages and applying DC gradients across the C-trap, ions can be bunched into short packets similar to those from the laser ion source. The C-trap is tightly integrated with the analyzer, injection optics and differential pumping.

Excitation

In principle, coherent axial oscillations of ion rings could be excited by applying RF waveforms to the outer electrode as demonstrated in[9] and references therein. However, if ion packets are injected away from the minimum of the axial potential (which corresponds to the thickest part of either electrode), this automatically initiates their axial oscillations, eliminating the need for any additional excitation. Furthermore, the absence of additional excitation allows the detection process to start as soon as the detection electronics recover from the voltage ramp needed for ion injection.

Detection

Axial oscillations of ion rings are detected by their image current induced on the outer electrode which is split into two symmetrical pick-up sensors connected to a differential amplifier. By processing data in a manner similar to that used in Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS), the trap can be used as a mass analyzer. Like in FTICR-MS, all the ions are detected simultaneously over some given period of time and resolution can be improved by increasing the strength of the field or by increasing the detection period. The Orbitrap differs from FTICR-MS by the absence of a magnetic field and hence has a significantly slower decrease of resolving power with increasing m/z.

Variants

Currently the Orbitrap analyzer exists in two variants: a standard trap and a compact high-field trap. In practical traps, the outer electrode is sustained at virtual ground and a voltage of 3.5 or 5 kV is applied to the inner electrode only. As a result, the resolving power at m/z 400 and 768 ms detection time can range from 60,000 for a standard trap at 3.5 kV to 280,000 for a high-field trap at 5 kV and with enhanced FT processing. Like in FTICR-MS the Orbitrap resolving power is proportional to the number of harmonic oscillations of the ions; as a result, the resolving power is inversely proportional to the square root of m/z and proportional to acquisition time. For example, the values above would double for m/z 100 and halve for m/z 1600. For the shortest transient of 96 ms these values would be reduced by 8 times, whereas a resolving power in excess of 1,000,000 has been demonstrated in 3-second transients.[10]

The Orbitrap analyzer can be interfaced to a linear ion trap (LTQ Orbitrap family of instruments), quadrupole mass filter (Q Exactive family) or directly to an ion source (Exactive instrument, all marketed by Thermo Fisher Scientific). In addition, a higher-energy collision cell can be appended to the C-trap, with the further addition of electron-transfer dissociation at its back.[11] Most of these instruments have atmospheric pressure ion sources though an intermediate-pressure MALDI source can also be used (MALDI LTQ Orbitrap). All of these instruments provide a high mass accuracy (<2–3 ppm with external calibrant and <1–2 ppm with internal), a high resolving power (up to 240,000 at m/z 400), a high dynamic range and high sensitivity.

Applications

Orbitrap-based mass spectrometers are used in proteomics[12] and are also used in life science mass spectrometry such as metabolism, metabolomics,[13] environmental,[14] food and safety analysis.[15] Most of them are interfaced to liquid chromatography separations,[14] though they are also used with gas chromatography,[16] secondary ion[17] and ambient ionization methods. They have also been used to determine molecular structures of isotopically substituted molecular species.[18]

See also

External links

Notes and References

  1. Makarov A . Electrostatic axially harmonic orbital trapping: a high-performance technique of mass analysis . Analytical Chemistry . 72 . 6 . 1156–1162 . March 2000 . 10740853 . 10.1021/ac991131p .
  2. Hu Q, Noll RJ, Li H, Makarov A, Hardman M, Graham Cooks R . The Orbitrap: a new mass spectrometer . Journal of Mass Spectrometry . 40 . 4 . 430–443 . April 2005 . 15838939 . 10.1002/jms.856 . 2005JMSp...40..430H .
  3. Kingdon KH . A Method for the Neutralization of Electron Space Charge by Positive Ionization at Very Low Gas Pressures. . 21 . 4 . 408–418 . 10.1103/PhysRev.21.408 . 1923 . 1923PhRv...21..408K .
  4. Storage of ions from laser-produced plasmas. Applied Physics Letters. 1981. Knight RD . 38. 4. 221–223. 10.1063/1.92315. 1981ApPhL..38..221K. live. https://web.archive.org/web/20151222223625/http://scitation.aip.org/content/aip/journal/apl/38/4/10.1063/1.92315. 22 December 2015. 2007-11-30.
  5. Web site: Памяти профессора Голикова Юрия Константиновича . In memory of Professor Golikov Yuri Konstantinovich . ru . Makarov A . 2005 . iairas.ru . 2023-10-13.
  6. Web site: Makarov A, Scigelova M . Orbitrap Mass Analyser: Overview and Applications in Proteomics . December 2014 . SlideServe .
  7. Makarov A, Denisov E, Kholomeev A, Balschun W, Lange O, Strupat K, Horning S . Performance evaluation of a hybrid linear ion trap/orbitrap mass spectrometer . Analytical Chemistry . 78 . 7 . 2113–2120 . April 2006 . 16579588 . 10.1021/ac0518811 .
  8. Makarov A, Denisov E, Lange O, Horning S . Dynamic range of mass accuracy in LTQ Orbitrap hybrid mass spectrometer . Journal of the American Society for Mass Spectrometry . 17 . 7 . 977–982 . July 2006 . 16750636 . 10.1016/j.jasms.2006.03.006 . free .
  9. Perry RH, Cooks RG, Noll RJ . Orbitrap mass spectrometry: instrumentation, ion motion and applications . Mass Spectrometry Reviews . 27 . 6 . 661–699 . 2008 . 18683895 . 10.1002/mas.20186 . 2008MSRv...27..661P .
  10. Web site: Orbitrap Mass Spectrometry With Resolving Powers Above 500,000 and 1,000,000 on a Chromatographic Time Scale. Denisov E, Damoc E, Makarov A, Lange O . Thermo Fisher Scientific. Bremen, Germany. 3 October 2020.
  11. McAlister GC, Berggren WT, Griep-Raming J, Horning S, Makarov A, Phanstiel D, Stafford G, Swaney DL, Syka JE, Zabrouskov V, Coon JJ . A proteomics grade electron transfer dissociation-enabled hybrid linear ion trap-orbitrap mass spectrometer . Journal of Proteome Research . 7 . 8 . 3127–3136 . August 2008 . 18613715 . 2601597 . 10.1021/pr800264t .
  12. Scigelova M, Makarov A . Orbitrap mass analyzer--overview and applications in proteomics . Proteomics . 6 . 16–21 . September 2006 . Suppl 2 . 17031791 . 10.1002/pmic.200600528 . 12774202 .
  13. Lu W, Clasquin MF, Melamud E, Amador-Noguez D, Caudy AA, Rabinowitz JD . Metabolomic analysis via reversed-phase ion-pairing liquid chromatography coupled to a stand alone orbitrap mass spectrometer . Analytical Chemistry . 82 . 8 . 3212–3221 . April 2010 . 20349993 . 2863137 . 10.1021/ac902837x .
  14. Wang J, Gardinali PR . Identification of phase II pharmaceutical metabolites in reclaimed water using high resolution benchtop Orbitrap mass spectrometry . Chemosphere . 107 . 65–73 . July 2014 . 24875872 . 10.1016/j.chemosphere.2014.03.021 . 2014Chmsp.107...65W .
  15. Makarov A, Scigelova M . Coupling liquid chromatography to Orbitrap mass spectrometry . Journal of Chromatography A . 1217 . 25 . 3938–3945 . June 2010 . 20299023 . 10.1016/j.chroma.2010.02.022 .
  16. Peterson AC, McAlister GC, Quarmby ST, Griep-Raming J, Coon JJ . Development and characterization of a GC-enabled QLT-Orbitrap for high-resolution and high-mass accuracy GC/MS . Analytical Chemistry . 82 . 20 . 8618–8628 . October 2010 . 20815337 . 10.1021/ac101757m .
  17. Passarelli MK, Pirkl A, Moellers R, Grinfeld D, Kollmer F, Havelund R, Newman CF, Marshall PS, Arlinghaus H, Alexander MR, West A, Horning S, Niehuis E, Makarov A, Dollery CT, Gilmore IS . The 3D OrbiSIMS-label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power . Nature Methods . 14 . 12 . 1175–1183 . December 2017 . 29131162 . 10.1038/nmeth.4504 . 54550356 .
  18. Eiler J, Cesar J, Chimiak L, Dallas B, Grice K, Griep-Raming J, Juchelka D, Kitchen N, Lloyd M, Makarov A, Robins R . Analysis of molecular isotopic structures at high precision and accuracy by Orbitrap mass spectrometry . International Journal of Mass Spectrometry . 2017 . 422 . 126–142 . 10.1016/j.ijms.2017.10.002. free . 2017IJMSp.422..126E .