FT-ICR MS inherently offers the highest broadband mass resolution and mass accuracy, partly because FT-ICR mass resolving power is equal to the measured frequency resolving power, whereas orbitrap or time-of-flight resolving power is the square root of the measured frequency or time-of-flight. FT-ICR mass resolving power varies directly with applied magnetic field strength (NHMFL’s 21 tesla offers the world’s highest field for ICR [1]), and may be tripled by segmented electrode detection. FT-ICR MS is uniquely advantageous for resolution and assignment of elemental compositions for complex mixtures, ranging from petroleum crude oil (>125,000 assigned peaks in a single mass spectrum) to top-down proteomics. MS/MS techniques include UV, multiphoton IR, collision-induced, and electron transfer dissociation [2], with additional enhancements by proton transfer, product ion “parking”, and ultrahigh-resolution precursor isolation by stored waveform inverse FT excitation [3]. Coupling with on-line HPLC can increase dynamic range by an order of magnitude. Benchmark performance will be illustrated for applications ranging from environmental oil spills to diagnostic identification of proteoforms in multiple myeloma and hemoglobinopathies. This work was performed at the National High Magnetic Field Laboratory, which is supported by NSF Division of Chemistry through DMR-1157490 and DMR-1644779 and the State of Florida.

FT-ICR MS inherently offers the highest broadband mass resolution and mass accuracy, partly because FT-ICR mass resolving power is equal to the measured frequency resolving power, whereas orbitrap or time-of-flight resolving power is the square root of the measured frequency or time-of-flight. FT-ICR mass resolving power varies directly with applied magnetic field strength (NHMFL’s 21 tesla offers the world’s highest field for ICR [1]), and may be tripled by segmented electrode detection. FT-ICR MS is uniquely advantageous for resolution and assignment of elemental compositions for complex mixtures, ranging from petroleum crude oil (>125,000 assigned peaks in a single mass spectrum) to top-down proteomics. MS/MS techniques include UV, multiphoton IR, collision-induced, and electron transfer dissociation [2], with additional enhancements by proton transfer, product ion “parking”, and ultrahigh-resolution precursor isolation by stored waveform inverse FT excitation [3]. Coupling with on-line HPLC can increase dynamic range by an order of magnitude. Benchmark performance will be illustrated for applications ranging from environmental oil spills to diagnostic identification of proteoforms in multiple myeloma and hemoglobinopathies. This work was performed at the National High Magnetic Field Laboratory, which is supported by NSF Division of Chemistry through DMR-1157490 and DMR-1644779 and the State of Florida.

FT-ICR MS inherently offers the highest broadband mass resolution and mass accuracy, partly because FT-ICR mass resolving power is equal to the measured frequency resolving power, whereas orbitrap or time-of-flight resolving power is the square root of the measured frequency or time-of-flight. FT-ICR mass resolving power varies directly with applied magnetic field strength (NHMFL’s 21 tesla offers the world’s highest field for ICR [1]), and may be tripled by segmented electrode detection. FT-ICR MS is uniquely advantageous for resolution and assignment of elemental compositions for complex mixtures, ranging from petroleum crude oil (>125,000 assigned peaks in a single mass spectrum) to top-down proteomics. MS/MS techniques include UV, multiphoton IR, collision-induced, and electron transfer dissociation [2], with additional enhancements by proton transfer, product ion “parking”, and ultrahigh-resolution precursor isolation by stored waveform inverse FT excitation [3]. Coupling with on-line HPLC can increase dynamic range by an order of magnitude. Benchmark performance will be illustrated for applications ranging from environmental oil spills to diagnostic identification of proteoforms in multiple myeloma and hemoglobinopathies. This work was performed at the National High Magnetic Field Laboratory, which is supported by NSF Division of Chemistry through DMR-1157490 and DMR-1644779 and the State of Florida.

Although mass information of all compounds is acquired at the same time leading to high throughput parallel acquisition, the structural information (MS/MS spectrum) is obtained sequentially leading to slow serial acquisition. We demonstrated that two-dimensional (2D) FT-ICR solve problem of obtaining all structural information as it records the correlations, in only one experiment, regardless of the complexity of the sample. 2D Fourier transform techniques have revolutionized nuclear magnetic resonance (NMR) since their introduction by R. R. Ernst in 1974. They paved the way for the analysis of complex samples such as purified proteins but also of complex mixtures such as plasma or urine. In the same way the principle of two-dimensional 2D FT-ICR MS was established in the late eighties inspired by the NOESY sequence. Unfortunately, 2D FT-ICR MS did not follow the rapid development of 2D NMR as its initial version suffered from three main drawbacks: (i) loss of resolution caused by poor vacuum induced by Collision Induced Dissociation in-cell fragmentation; (ii) difficulty in data treatment at full FT-ICR resolution and (iii) intense scintillation noise. During the last years, we revisited 2D FT-ICR MS with modern instrumentation using gas-free fragmentation methods such as IRMPD and ECD, introduced optimized pulse sequence and advanced data processing. With these developments, 2D FT-ICR MS has been able to provide structural information on samples of increasing complexity on all fields of analytical chemistry. Nevertheless, until now 2D FT-ICR was hampered by the limited resolution for precursor ion selection of the order of several mass units. An overnight acquisition is already required to reach a quadrupole mass filter-like unit mass resolution. We have recently reported that 2D FT-ICR MS using non-uniform sampling (NUS) obtained by randomly skipping points in the first dimension corresponding to the precursor selection gives access, after data processing, to the same structural information contained in a complex mixture. The resolution increases roughly as the inverse of the NUS ratio, up to roughly 32 times at NUS 1/32, leading to an acquisition with resolution of 10,000 for the precursor ions at m/z 200 in less than one hour. Examples will be presented in different fields (proteomics, lipidomics, metabolomics) applied to heritage and archaeological samples of 1D and 2D FT-ICR MS.

Although mass information of all compounds is acquired at the same time leading to high throughput parallel acquisition, the structural information (MS/MS spectrum) is obtained sequentially leading to slow serial acquisition. We demonstrated that two-dimensional (2D) FT-ICR solve problem of obtaining all structural information as it records the correlations, in only one experiment, regardless of the complexity of the sample. 2D Fourier transform techniques have revolutionized nuclear magnetic resonance (NMR) since their introduction by R. R. Ernst in 1974. They paved the way for the analysis of complex samples such as purified proteins but also of complex mixtures such as plasma or urine. In the same way the principle of two-dimensional 2D FT-ICR MS was established in the late eighties inspired by the NOESY sequence. Unfortunately, 2D FT-ICR MS did not follow the rapid development of 2D NMR as its initial version suffered from three main drawbacks: (i) loss of resolution caused by poor vacuum induced by Collision Induced Dissociation in-cell fragmentation; (ii) difficulty in data treatment at full FT-ICR resolution and (iii) intense scintillation noise. During the last years, we revisited 2D FT-ICR MS with modern instrumentation using gas-free fragmentation methods such as IRMPD and ECD, introduced optimized pulse sequence and advanced data processing. With these developments, 2D FT-ICR MS has been able to provide structural information on samples of increasing complexity on all fields of analytical chemistry. Nevertheless, until now 2D FT-ICR was hampered by the limited resolution for precursor ion selection of the order of several mass units. An overnight acquisition is already required to reach a quadrupole mass filter-like unit mass resolution. We have recently reported that 2D FT-ICR MS using non-uniform sampling (NUS) obtained by randomly skipping points in the first dimension corresponding to the precursor selection gives access, after data processing, to the same structural information contained in a complex mixture. The resolution increases roughly as the inverse of the NUS ratio, up to roughly 32 times at NUS 1/32, leading to an acquisition with resolution of 10,000 for the precursor ions at m/z 200 in less than one hour. Examples will be presented in different fields (proteomics, lipidomics, metabolomics) applied to heritage and archaeological samples of 1D and 2D FT-ICR MS.

Although mass information of all compounds is acquired at the same time leading to high throughput parallel acquisition, the structural information (MS/MS spectrum) is obtained sequentially leading to slow serial acquisition. We demonstrated that two-dimensional (2D) FT-ICR solve problem of obtaining all structural information as it records the correlations, in only one experiment, regardless of the complexity of the sample. 2D Fourier transform techniques have revolutionized nuclear magnetic resonance (NMR) since their introduction by R. R. Ernst in 1974. They paved the way for the analysis of complex samples such as purified proteins but also of complex mixtures such as plasma or urine. In the same way the principle of two-dimensional 2D FT-ICR MS was established in the late eighties inspired by the NOESY sequence. Unfortunately, 2D FT-ICR MS did not follow the rapid development of 2D NMR as its initial version suffered from three main drawbacks: (i) loss of resolution caused by poor vacuum induced by Collision Induced Dissociation in-cell fragmentation; (ii) difficulty in data treatment at full FT-ICR resolution and (iii) intense scintillation noise. During the last years, we revisited 2D FT-ICR MS with modern instrumentation using gas-free fragmentation methods such as IRMPD and ECD, introduced optimized pulse sequence and advanced data processing. With these developments, 2D FT-ICR MS has been able to provide structural information on samples of increasing complexity on all fields of analytical chemistry. Nevertheless, until now 2D FT-ICR was hampered by the limited resolution for precursor ion selection of the order of several mass units. An overnight acquisition is already required to reach a quadrupole mass filter-like unit mass resolution. We have recently reported that 2D FT-ICR MS using non-uniform sampling (NUS) obtained by randomly skipping points in the first dimension corresponding to the precursor selection gives access, after data processing, to the same structural information contained in a complex mixture. The resolution increases roughly as the inverse of the NUS ratio, up to roughly 32 times at NUS 1/32, leading to an acquisition with resolution of 10,000 for the precursor ions at m/z 200 in less than one hour. Examples will be presented in different fields (proteomics, lipidomics, metabolomics) applied to heritage and archaeological samples of 1D and 2D FT-ICR MS.

The molecular description of heavy petroleum fractions, such as vacuum gas oils or bitumen, remains to be one of the most challenging analytical tasks. These fractions are extremely complex mixtures, exhibiting various chemical functionalities, with low volatility and solubility. Commonly, direct liquid infusion techniques are deployed, such as electrospray or atmospheric pressure photoionization (APPI). The hyphenation of thermal analysis techniques, by means of evolved gas analysis, is one complementary approach for deciphering these materials. The controlled heating of the sample material allows for the specification of the desorbed material at lower temperatures, whereas the induced pyrolysis at elevated temperatures allows for conclusions on building blocks. Two thermal analysis strategies hyphenated to high-resolution mass spectrometry are shown: 1) Atmospheric pressure (AP) thermogravimetry coupled to APPI and atmospheric pressure chemical ionization (APCI) ultra-high resolution Fourier transform mass spectrometry (FTICR MS), and 2) AP direct inlet probe (DIP) coupled to APCI/APPI FTICR MS. Apart from various crude oil samples and their distillation fractions, such as vacuum gas oil, bitumen samples (aged and polymeric mixtures) were in focus of the work. Data fusion strategies to combine the results from the individual techniques are presented utilizing self-written Matlab scripts and processing algorithms to allow for a comprehensive data treatment of the time/temperature-resolved high-resolution mass spectra. Each of the deployed thermal analysis strategies offers unique possibilities. The thermogravimetric coupling is able to detect the mass loss during the defined heating procedure very sensitively and is able to reach the highest temperatures (> 1000 °C), whereas direct inlet probe coupling with a maximum temperature of 450 °C enables to conduct measurements very rapidly (5-10 min). Atmospheric pressure DIP turned out to be the most simple of the presented techniques and was operated in two modes: direct 400 °C evaporation and fast-temperature ramp (20-50 K/min) analysis. The later allows acquiring more defined temperature-profiles. In particular, for the bitumen-polymer mixtures, a slow ramp revealed to be beneficial and significant changes between the evolved gas patterns of the different formulations were observed. Finally, none of the individual techniques gave a comprehensive picture of the petroleomic materials enforcing data fusion strategies. Preliminary attempts revealed a significant enlargement of covered chemical space when combining the data.

The molecular description of heavy petroleum fractions, such as vacuum gas oils or bitumen, remains to be one of the most challenging analytical tasks. These fractions are extremely complex mixtures, exhibiting various chemical functionalities, with low volatility and solubility. Commonly, direct liquid infusion techniques are deployed, such as electrospray or atmospheric pressure photoionization (APPI). The hyphenation of thermal analysis techniques, by means of evolved gas analysis, is one complementary approach for deciphering these materials. The controlled heating of the sample material allows for the specification of the desorbed material at lower temperatures, whereas the induced pyrolysis at elevated temperatures allows for conclusions on building blocks. Two thermal analysis strategies hyphenated to high-resolution mass spectrometry are shown: 1) Atmospheric pressure (AP) thermogravimetry coupled to APPI and atmospheric pressure chemical ionization (APCI) ultra-high resolution Fourier transform mass spectrometry (FTICR MS), and 2) AP direct inlet probe (DIP) coupled to APCI/APPI FTICR MS. Apart from various crude oil samples and their distillation fractions, such as vacuum gas oil, bitumen samples (aged and polymeric mixtures) were in focus of the work. Data fusion strategies to combine the results from the individual techniques are presented utilizing self-written Matlab scripts and processing algorithms to allow for a comprehensive data treatment of the time/temperature-resolved high-resolution mass spectra. Each of the deployed thermal analysis strategies offers unique possibilities. The thermogravimetric coupling is able to detect the mass loss during the defined heating procedure very sensitively and is able to reach the highest temperatures (> 1000 °C), whereas direct inlet probe coupling with a maximum temperature of 450 °C enables to conduct measurements very rapidly (5-10 min). Atmospheric pressure DIP turned out to be the most simple of the presented techniques and was operated in two modes: direct 400 °C evaporation and fast-temperature ramp (20-50 K/min) analysis. The later allows acquiring more defined temperature-profiles. In particular, for the bitumen-polymer mixtures, a slow ramp revealed to be beneficial and significant changes between the evolved gas patterns of the different formulations were observed. Finally, none of the individual techniques gave a comprehensive picture of the petroleomic materials enforcing data fusion strategies. Preliminary attempts revealed a significant enlargement of covered chemical space when combining the data.

The molecular description of heavy petroleum fractions, such as vacuum gas oils or bitumen, remains to be one of the most challenging analytical tasks. These fractions are extremely complex mixtures, exhibiting various chemical functionalities, with low volatility and solubility. Commonly, direct liquid infusion techniques are deployed, such as electrospray or atmospheric pressure photoionization (APPI). The hyphenation of thermal analysis techniques, by means of evolved gas analysis, is one complementary approach for deciphering these materials. The controlled heating of the sample material allows for the specification of the desorbed material at lower temperatures, whereas the induced pyrolysis at elevated temperatures allows for conclusions on building blocks. Two thermal analysis strategies hyphenated to high-resolution mass spectrometry are shown: 1) Atmospheric pressure (AP) thermogravimetry coupled to APPI and atmospheric pressure chemical ionization (APCI) ultra-high resolution Fourier transform mass spectrometry (FTICR MS), and 2) AP direct inlet probe (DIP) coupled to APCI/APPI FTICR MS. Apart from various crude oil samples and their distillation fractions, such as vacuum gas oil, bitumen samples (aged and polymeric mixtures) were in focus of the work. Data fusion strategies to combine the results from the individual techniques are presented utilizing self-written Matlab scripts and processing algorithms to allow for a comprehensive data treatment of the time/temperature-resolved high-resolution mass spectra. Each of the deployed thermal analysis strategies offers unique possibilities. The thermogravimetric coupling is able to detect the mass loss during the defined heating procedure very sensitively and is able to reach the highest temperatures (> 1000 °C), whereas direct inlet probe coupling with a maximum temperature of 450 °C enables to conduct measurements very rapidly (5-10 min). Atmospheric pressure DIP turned out to be the most simple of the presented techniques and was operated in two modes: direct 400 °C evaporation and fast-temperature ramp (20-50 K/min) analysis. The later allows acquiring more defined temperature-profiles. In particular, for the bitumen-polymer mixtures, a slow ramp revealed to be beneficial and significant changes between the evolved gas patterns of the different formulations were observed. Finally, none of the individual techniques gave a comprehensive picture of the petroleomic materials enforcing data fusion strategies. Preliminary attempts revealed a significant enlargement of covered chemical space when combining the data.

Studying chemical transformations within the carbon space in complex mixtures is a growing research field. In different areas such as metabolomics, environmental studies or medical areas, the complexity of chemical problems is increasing. To be able to understand chemical or biological transformations on a molecular level high performance analytical systems are necessary, with mass spectrometry leading the way. While the best possible performance regarding key factors mass resolution, mass accuracy and sensitivity is always desirable, the combination of all three parameters becomes crucial when dealing with highly complex samples. High-field Orbitrap mass spectrometer show high resolution and accuracy with a much improved sensitivity compared to FT-ICR MS while reducing analysis time. Here, the faster transients and shorter ion transfer are key points that favor the Orbitrap. For a detailed study of complex systems using ultrahigh resolution MS, a heavy crude oil was used to characterize the capabilities of highfield Orbitrap using an APPI source. Analyses where performed on a research-type Orbitrap Elite (Thermo Fisher, Bremen, Germany) at resolving powers of 240k, 480k and 960k (@ m/z 400) by employing both full range scanning (200-1200 Da) and spectral stitching (30 Da windows, 5 Da overlap). The detailed MS analysis of complex mixtures suffers from the need to resolve isobaric signals. In crude oil one of the most crucial mass splits to cover is the 3.4 mDa difference between C3 and SH4. The instrument used here allows a resolving power of 960k at 400 Da (500k at 1000 Da). An approximately 1.5-fold increase of assigned signals is observed for each doubling of the resolving power set. Mass spectral stitching leads to an increase of assignable signals by a factor of 6 to 9. Considering the data achieved by FTOrbitrap MS essentially all potential assignments are found that are theoretical possible in a certain mass range for different compound classes, thus covering a large part of the carbon space.

Studying chemical transformations within the carbon space in complex mixtures is a growing research field. In different areas such as metabolomics, environmental studies or medical areas, the complexity of chemical problems is increasing. To be able to understand chemical or biological transformations on a molecular level high performance analytical systems are necessary, with mass spectrometry leading the way. While the best possible performance regarding key factors mass resolution, mass accuracy and sensitivity is always desirable, the combination of all three parameters becomes crucial when dealing with highly complex samples. High-field Orbitrap mass spectrometer show high resolution and accuracy with a much improved sensitivity compared to FT-ICR MS while reducing analysis time. Here, the faster transients and shorter ion transfer are key points that favor the Orbitrap. For a detailed study of complex systems using ultrahigh resolution MS, a heavy crude oil was used to characterize the capabilities of highfield Orbitrap using an APPI source. Analyses where performed on a research-type Orbitrap Elite (Thermo Fisher, Bremen, Germany) at resolving powers of 240k, 480k and 960k (@ m/z 400) by employing both full range scanning (200-1200 Da) and spectral stitching (30 Da windows, 5 Da overlap). The detailed MS analysis of complex mixtures suffers from the need to resolve isobaric signals. In crude oil one of the most crucial mass splits to cover is the 3.4 mDa difference between C3 and SH4. The instrument used here allows a resolving power of 960k at 400 Da (500k at 1000 Da). An approximately 1.5-fold increase of assigned signals is observed for each doubling of the resolving power set. Mass spectral stitching leads to an increase of assignable signals by a factor of 6 to 9. Considering the data achieved by FTOrbitrap MS essentially all potential assignments are found that are theoretical possible in a certain mass range for different compound classes, thus covering a large part of the carbon space.

Studying chemical transformations within the carbon space in complex mixtures is a growing research field. In different areas such as metabolomics, environmental studies or medical areas, the complexity of chemical problems is increasing. To be able to understand chemical or biological transformations on a molecular level high performance analytical systems are necessary, with mass spectrometry leading the way. While the best possible performance regarding key factors mass resolution, mass accuracy and sensitivity is always desirable, the combination of all three parameters becomes crucial when dealing with highly complex samples. High-field Orbitrap mass spectrometer show high resolution and accuracy with a much improved sensitivity compared to FT-ICR MS while reducing analysis time. Here, the faster transients and shorter ion transfer are key points that favor the Orbitrap. For a detailed study of complex systems using ultrahigh resolution MS, a heavy crude oil was used to characterize the capabilities of highfield Orbitrap using an APPI source. Analyses where performed on a research-type Orbitrap Elite (Thermo Fisher, Bremen, Germany) at resolving powers of 240k, 480k and 960k (@ m/z 400) by employing both full range scanning (200-1200 Da) and spectral stitching (30 Da windows, 5 Da overlap). The detailed MS analysis of complex mixtures suffers from the need to resolve isobaric signals. In crude oil one of the most crucial mass splits to cover is the 3.4 mDa difference between C3 and SH4. The instrument used here allows a resolving power of 960k at 400 Da (500k at 1000 Da). An approximately 1.5-fold increase of assigned signals is observed for each doubling of the resolving power set. Mass spectral stitching leads to an increase of assignable signals by a factor of 6 to 9. Considering the data achieved by FTOrbitrap MS essentially all potential assignments are found that are theoretical possible in a certain mass range for different compound classes, thus covering a large part of the carbon space.