Whatever the Wavelength: The Wide World of Spectroscopy

In 1995, the Australian Marc Wilkins characterized the concept of the proteome as the “full protein equivalent of a genome”. In relation to metabolic products, the emerging research field of “proteomics” soon found its equivalent in “metabolomics”. Both approaches provide much more information than “genomics” alone, from which everything originated. Through to today, the trend towards analyzing a wide range of bioanalytical or medical issues “holistically” in this way has continued unabated. Applications range from basic research in medicine and the natural sciences, through diagnostics, to applications in doping analytics or food control. The possibility of conducting both qualitative and quantitative investigations and its high resolution, mass accuracy, selectivity and sensitivity make mass spectrometry ideally suited for these new areas of application. At the Bremen site of Thermo Electron, the first step was taken towards Life Science mass spectrometry in 2003, with the introduction of the LTQ-FT Fourier Transform mass spectrometer.

Orbitrap: a milestone in analyzer technology

A true milestone followed in 2005, with a further FT-MS detector: the Orbitrap, in a system known as the LTQ Orbitrap. This new analyzer type achieved revolutionary data in terms of resolution and mass accuracy. How? To simplify things somewhat, by measuring frequencies with which ions migrate around a central, spindle-shaped electrode in an electrostatic field, depending on their mass-to-charge ratio. “Within physical properties, frequencies are the most accurate thing you can measure,” explains Dr. Thomas Moehring, Senior Director Hardware Management, Life Science Mass Spectrometry, at Thermo Fisher Scientific’s Bremen site. “The resolution is directly proportional to the time during which we are able to cause the ions to migrate in the analyzer. The longer they migrate, the more data points and therefore the higher the resolution.”

Milestone Spectroscopy

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The scientific community was impressed — and was quickly calling for more: more resolution in a shorter time, i.e. more speed. As an initial response to that, Thermo Fisher revised the geometry of the Orbitrap analyzer and introduced its second generation, known as the High-Field Orbitrap. From then on, various product families were continuously further developed, adapted to the respective requirements and markets — and all having the Orbitrap as analyzer in common. The benchtop systems of the Q Exactive MS family with Q Exactive, Q Exactive Plus, Q Exactive HF and Q Exactive HF X-Hybrid Quadrupole Orbitrap mass spectrometers became the workhorses for proteome analysis.

The high-end research market is serviced primarily by systems such as the Orbitrap Fusion Lumos from the Tribrid product group, and Q Exactive HF and HF-X. In the field of proteomics, they cover every experiment for protein analysis, starting from basic experiments such as peptide ID and protein ID through database searches in relation to tryptic digestion to quantifications and characterizations of intact antibodies, fragmentations of antibodies or characterizations of post-translational modifications (PTMs).

The latest system from the benchtop Q Exactive MS family, the Q Exactive-HF-X hybrid quadrupole Orbitrap mass spectrometer, combines a modern segmented quadrupole for high-performance precursor ion selection with the high-field Orbitrap. Its low detection limits cover a range from small molecules to the native antibody. Compared to the Q Exactive and Q Exactive HF models, the analysis time in this system was again reduced by a factor of 2, whilst maintaining high robustness and reproducibility, thus permitting higher throughputs.

Needs-matched development of hardware & software

Nowadays, Orbitrap technology has also penetrated a wide range of applied markets, such as food analysis or environmental analysis. These entail very different requirements in some cases — for both hardware and software. Thomas Moehring comments: “Instrument Control Software has advanced massively in recent years. From “research-like”, where customers were able to adjust all buttons, all lenses and all voltages, to more market-oriented method templates and a simplification of the parameters still needing to be played around with. User-friendliness and the user experience are key buzzwords here.

“In 2013, we introduced the first Tribrid device, featuring a triple and an Orbitrap on an Instrument Control Software platform for the first time ever. This was the first step towards a harmonized platform, and that is also set to be the strategy we will be pursuing in future. Depending on the market being targeted, with different device types, specific pre-prepared methods and a certain degree of flexibility, or equally non-flexibility”.

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