15N isotope-selective infrared multiphoton dissociation of nitromethane by a free electron laser

Successful experiments on the isotope-selective infrared multiphoton dissociation (IR MPD) of nitromethane molecules in the region of stretching vibrations of the NO₂ group have been performed for the first time under IR free electron laser (FEL) irradiation in a mixture with the natural content of the¹⁵N isotope of 0.4%. The content of the¹⁵N isotope in the products of NO dissociation varies within 0.1–1.6% as a function of the laser radiation frequency. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 4, pp. 739–742, April, 1998.     

Study on the feasibility of obtaining highly enriched Carbon-13 in one- and two- stage modes of laser separation process

The feasibility of obtaining highly enriched (¹³C & 80%) carbon-13 by isotope-selective IR multiphoton dissociation (MPD) of Freon 22 (CF₂HCl) molecules in one- and two-stage modes of the process in an apparatus with the intracavity arrangement of the separation reactor, which resembled current technological facilities in its basic characteristics, was studied. The one-stage separation scheme was realized using the selective MPD of Freon 22 by which¹³C is concentrated in the dissociation product tetrafluoroethylene (C²F⁴). Carbon-13 concentrations in the product and MPD yields were measured depending on the laser radiation frequency and Freon 22 and bath gas (nitrogen) pressure. At the chosen irradiation geometry and a laser pulse repetition rate of 30 Hz,¹³C concentrations of 83 and 89% in the final product with the production rate of ∼36 and ∼7 mg¹³C/h were attained. The two-stage separation scheme was furnished on the basis of isotopically selective dissociation of the intermediate product CF²HI enriched in¹³C by its buildup process upon irradiation of a CF²HCl mixture with HI. The final product in this case was CF²H² having a¹³C concentration of 98 ± 1.5%. The production rate of the two-stage laser separation process was ∼22 mg¹³C/h.     

Collisionally assisted, highly selective laser isotope separation of carbon-13

We have further developed our recently reported two-laser technique for highly selective molecular isotope separation of carbon-13 [Boyarkin, Kowalczyk, and Rizzo, J. Chem. Phys. 118, 93 (2003)] with the objective of increasing the yield. An essential feature of this approach in its original conception is the significant increase of isotopic selectivity that occurs through collisions during the time between the overtone preexcitation laser pulse and the multiphoton dissociation pulse. We demonstrate here that under certain conditions, this collisional enhancement of the selectivity works equally well when the two pulses are overlapped in time, allowing the overall isotopic selectivity of the process to remain high while achieving a significant increase in the absolute dissociation yield. We also find that proper shaping of the CO2 laser dissociation pulse makes the fluence required for dissociation sufficiently low to allow irradiation of a large reaction volume by unfocused laser beams. Together, these factors may make this laser isotope separation scheme competitive with existing separation methods.     

Isotopically selective infrared multiphoton dissociation of 2,3-dihydropyran

Oxygen isotopic selectivity on infrared multiphoton dissociation of 2,3-dihydropyran has been studied by the examination of the effects of excitation frequency, laser fluence, and gas pressure on the dissociation probability of 2,3-dihydropyran and isotopic composition of products. Oxygen-18 was enriched in a dissociation product: 2-propenal. The enrichment factor of 18O and the dissociation probability were measured at a laser frequency between 1033.5 and 1057.3 cm-1, the laser fluence of 2.2-2.3 J/cm2, and the 2,3-dihydropyran pressure of 0.27 kPa. The dissociation probability decreases as the laser frequency being detuned from the absorption peak of 2,3-dihydropyran around 1081 cm-1. On the other hand, the enrichment factor increases with detuning the frequency. The enrichment factor of 18O increases with increasing the 2,3-dihydropyran pressure at the laser fluence of 2.7 J/cm2 or less and the laser frequency of 1033.5 cm-1, whereas the yield of 2-propenal decreases with increasing the pressure. A very high enrichment factor of 751 was obtained by the irradiation of 0.53 kPa of 2,3-dihydropyran at 2.1 J/cm2. Collisional effect of vibrationally excited molecules with ambient molecules on isotopic selectivity is discussed on the basis of a rate equation model including a collisional vibrational de-excitation process.     

Study on the Capabilities of One- and Two-Stage Separation Schemes for Producing Highly Enriched Carbon-13 by the Laser Method

The feasibility of obtaining highly enriched (up to 99.5%) carbon-13 by the method of isotopically selective IR multiple-photon dissociation (MPD) of Freon 22 (CF₂HCl) molecules was studied for one- and two-stage schemes of the process in a laboratory apparatus with the intracavity location of a separation reactor. The concentration of ¹³C in the dissociation product C₂F₄ (tetrafluoroethylene) and the MPD yields were measured depending on the frequency and fluence of laser radiation, as well as on the Freon 22 pressure. At the irradiation geometry used, the concentrations of ¹³C up to 88% were obtained in the first stage of enrichment. The second enrichment stage was based on the isotope-selective dissociation of Freon 22 preliminarily enriched with ¹³C (up to 32.6%). After the second stage, the final ¹³C concentration of 99.5 ± 0.5% was attained in the product C₂F₄.     

Infrared multiphoton dissociation of small-interfering RNA anions and cations

Infrared multiphoton dissociation (IRMPD) on a linear ion trap mass spectrometer is applied for the sequencing of small interfering RNA (siRNA). Both single-strand siRNAs and duplex siRNA were characterized by IRMPD, and the results were compared with that obtained by traditional ion trap-based collision induced dissociation (CID). The single-strand siRNA anions were observed to dissociate via cleavage of the 5′ P—O bonds yielding c- and y-type product ions as well as undergo neutral base loss. Full sequence coverage of the siRNA anions was obtained by both IRMPD and CID. While the CID mass spectra were dominated by base loss ions, accounting for ∼25% to 40% of the product ion current, these ions were eliminated through secondary dissociation by increasing the irradiation time in the IRMPD mass spectra to produce higher abundances of informative sequence ions. With longer irradiation times, however, internal ions corresponding to cleavage of two 5′ P—O bonds began to populate the product ion mass spectra as well as higher abundances of [a − Base] and w-type ions. IRMPD of siRNA cations predominantly produced c- and y-type ions with minimal contributions of [a − Base] and w-type ions to the product ion current; the presence of only two complementary series of product ions in the IRMPD mass spectra simplified spectral interpretation. In addition, IRMPD produced high abundances of protonated nucleobases, [G + H]⁺, [A + H]⁺, and [C + H]⁺, which were not detected in the CID mass spectra due to the low-mass cut-off associated with conventional CID in ion traps. CID and IRMPD using short irradiation times of duplex siRNA resulted in strand separation, similar to the dissociation trends observed for duplex DNA. With longer irradiation times, however, the individual single-strands underwent secondary dissociation to yield informative sequence ions not obtained by CID.     

13C separation by IRMPD of CF2HCL molecules with a powerful CO2-TEA laser

The single pulse separation parameters in IRMPD of CF₂HCL molecules have been obtained in relation with the laser fluence, the number of laser pulses, and the gas pressure. The measurements have been carried out using a fixed laser line (9P22) and an ordinary shape of the laser pulses.     

Infrared absorption features of gaseous isopropyl carbocations.

C3H7+ ions were formed in the cell of a Fourier transform ion cyclotron resonance mass spectrometer and assayed by their multi-photon dissociation (MPD) behavior, triggered by the absorption of tunable IR radiation from a free-electron laser source providing a high fluence. The derived experimental IRMPD spectrum, which reflects the active vibrational modes of the ion, was compared with the IR spectra calculated for the optimized structures of the most-stable species on the C3H7+ potential energy surface, namely, a chiral iC3H7+ ion of C2 symmetry and an asymmetric corner-protonated cyclopropane, cC3H7+. The significant features in the IRMPD spectra of both the unlabeled and the perdeuterated ions obtained by ionization and fragmentation of isobutane or 2-chloro[D7]propane confirm the presence of the isopropyl cation, the ground-state isomer, whose IR spectroscopic features can thus be comparatively checked in the gas phase and in condensed superacid media. Details of the IRMPD features are suggested to result from the nearly barrierless interconversion of the two C2 enantiomers.     

Improving IRMPD in a quadrupole ion trap

A focused laser is used to make infrared multiphoton photodissociation (IRMPD) more efficient in a quadrupole ion trap mass spectrometer. Efficient (up to 100%) dissociation at the standard operating pressure of 1 × 10⁻³ Torr can be achieved without any supplemental ion activation and with shorter irradiation times. The axial amplitudes of trapped ion clouds are measured using laser tomography. Laser flux on the ion cloud is increased six times by focusing the laser so that the beam waist approximates the ion cloud size. Unmodified peptide ions from 200 Da to 3 kDa are completely dissociated in 2.5–10 ms at a bath gas pressure of 3.3 × 10⁻⁴ Torr and in 3–25 ms at 1.0 × 10⁻³ Torr. Sequential dissociation of product ions is increased by focusing the laser and by operating at an increased bath gas pressure to minimize the size of the ion cloud.     

Yield and enrichment studies of C-13 isotope by multi-photon dissociation of Freon-22 at low temperatures

Multi-photon dissociation of Freon-22 (CF₂HC1) at low temperatures has been carried out to separate the C-13 isotope using a TEA CO₂ laser. Yield and enrichment of C-13 isotope in the product C₂F₄ are studied at 9P(22) laser line as a function of temperature (−50°C to 30°C). It is observed that at a given fluence when the temperature is lowered the yield decreases and the enrichment factor of C-13 increases. Room temperature irradiation of CF₂HC1 towards the blue edge of C-13 absorption (i.e. at 9P(20) laser line) gives low yield of the product (C₂F₄) at a fluence, which produces the desired enrichment factor of 100. An increase in fluence gives very high yield of C₂F₄ but the enrichment factor is very low. Irradiating CF₂HC1 at a temperature of −10°C enhances the enrichment factor to 100 and the yield obtained is comparable to that towards the red edge of C-13 absorption (i.e. at 9P(26) laser line). At a given enrichment factor higher enrichment efficiency is achieved when CF₂HC1 is irradiated at lower temperature.     

CF2HCl分子在红外多光子解离时分子间的能量传递过程

The infrared multiphoton dissociation (IR MPD) of CF_2HCl by measuring the C_2F_4 product after laser irradiation is investigated, The dissociation fraction of CF_2HCl as a function of the CF_2HCl pressure, laser energy fluence and number of laser pulses is measured. Under the low pressure condition the bottleneck effect is observed. Under the collision condition the dissociation fraction of CF_2HCl is affected by the competition between rotaional hole filling (collisional repoputation of pumped rotational states) and collisional deactivation of excited CF_2HCl moleculae.     

Maximization of yield of C-13 isotope by multiphoton dissociation of Freon-22 using high average power TEA CO2 laser

Selective multi-photon dissociation (MPD) of Freon-22 (CF₂HC1) molecules has been carried out using a TEA CO₂ laser at various CO₂ laser lines (9P(20)-9P(26)) in order to maximize the yield of C-13 isotope in the product (C₂F₄) at an enrichment factor of 100. The effects of laser pulse tail due to the presence of N₂ in the laser mixture on the enrichment factor and yield of C-13 are investigated. It is found that the addition of a small amount of N₂ is possible in the laser mixture without a significant drop in the yield at desired enrichment factor. Addition of a small amount of N₂ improves the laser efficiency considerably. At a given pulse energy, a slight change in the near field intensity distribution of a laser severely affects the selectivity of C-13 isotope. The computed far-field intensity distributions of the measured near-field intensities show marked spatial variation in the focal spots that leads to a drop in selectivity. For macroscopic production of C-13 isotope a simple and novel multi-pass cavity has been designed and tested to focus the energy repeatedly keeping the optimum fluence constant at each focal spot.     

Combined infrared multiphoton dissociation and electron capture dissociation with a hollow electron beam in Fourier transform ion cyclotron resonance mass spectrometry

An electron injection system based on an indirectly heated ring-shaped dispenser cathode has been developed and installed in a 7 Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. This new hardware design allows high-rate electron capture dissociation (ECD) to be carried out by a hollow electron beam coaxial with the ion cyclotron resonance (ICR) trap. Infrared multiphoton dissociation (IRMPD) can also be performed with an on-axis IR-laser beam passing through a hole at the centre of the dispenser cathode. Electron and photon irradiation times of the order of 100 ms are required for efficient ECD and IRMPD, respectively. As ECD and IRMPD generate fragments of different types (mostly c, z and b, y, respectively), complementary structural information that improves the characterization of peptides and proteins by FTICR mass spectrometry can be obtained. The developed technique enables the consecutive or simultaneous use of the ECD and IRMPD methods within a single FTICR experimental sequence and on the same ensemble of trapped ions in multistage tandem (MS/MS/MS or MS(n)) mass spectrometry. Flexible changing between ECD and IRMPD should present advantages for the analysis of protein digests separated by liquid chromatography prior to FTICRMS. Furthermore, ion activation by either electron or laser irradiation prior to, as well as after, dissociation by IRMPD or ECD increases the efficiency of ion fragmentation, including the w-type fragment ion formation, and improves sequencing of peptides with multiple disulfide bridges. The developed instrumental configuration is essential for combined ECD and IRMPD on FTICR mass spectrometers with limited access into the ICR trap.     

Isotopically selective collisional vibrational energy transfer in CF3H

The authors investigate here the mechanism of collisionally enhanced isotopic selectivity observed in infrared multiple photon dissociation (IRMPD) of vibrationally preexcited CF3H by Boyarkin et al. [J. Chem. Phys. 118, 93 (2003)]. For both the carbon-12 and carbon-13 isotopic species they measure the dependence of the IRMPD yield on the time delay between the preexcitation and the dissociation pulses at different dissociation frequencies as well as its dependence on the initial isotopic composition of the sample. The results reveal that the collisional increase in isotopic selectivity originates not only from that of IRMPD itself but also from the isotopic selectivity of vibrational energy transfer, with the latter making the major contribution under their experimental conditions. They suggest that the observed isotopic selectivity in collisional energy transfer arises from the difference in overlap between the absorption spectra of the nu5 mode in the 12CF3H acceptor molecule with emission spectra of the same mode in the two isotopically different donors. Understanding the origin of this collisional effect has important implications for optimization of laser isotope separation processes.     

Photoinduced dissociation of electrospray-generated ions in an ion trap/time-of-flight mass spectrometer using a pulsed CO2 laser

An ion trap/time-of-flight (IT/TOF) mass spectrometer was developed and applied to infrared multiphoton dissociation (IRMPD) studies of ions generated by electrospray ionization. A pulsed 10.6- micro m laser beam from a CO(2) laser was used for excitation of trapped ions. Results from IRMPD of peptide ions show that this method provides useful information related to the amino acid sequence of analyzed peptides. Comparative studies show that IRMPD spectra are similar to those obtained using a 266-nm UV laser beam for excitation. However, in contrast to multiple-pulse excitation required at 266 nm, the energy of a single laser pulse in IRMPD is sufficient to induce dissociation of peptide ions. The laser power is practically an exclusive parameter that must be controlled in order to obtain IRMPD spectra that will provide the optimal structural information. It is further demonstrated that the IRMPD IT/TOF technique has the potential to probe the structural features of larger ions that cannot be readily fragmented by collision-induced dissociation (CID). A multiply charged ion of equine cytochrome c is successfully fragmented in a single laser pulse experiment. The IRMPD IT/TOF technique is also shown to be a promising tool for studying dissociation kinetics of peptide and protein ions. Unlike other methods that usually monitor the dissociation ion kinetics in a dissociation time frame of greater than milliseconds, the IT/TOF can promptly detect all product ions generated by the dissociation process, and thus monitor the dissociation process of peptides and proteins in a sub-millisecond time frame. This instrument allows us to determine the dissociation rates of cytochrome c ions using high-energy photoexcitation. It is found that the charge state of the protein ion has a significant effect on dissociation kinetics, which is consistent with that found under low-energy excitation experiments. It is shown that the increase in energy of a laser pulse from 130 to 180 mJ changes the dissociation rate constant for the +12 ion from k = 2.4 x 10(3) x s(-1) to k = 7.3 x 10(4) x s(-1). The +8 ion following excitation at 130 mJ dissociates slower with a rate constant of k = 2.6 x 10(2) x s(-1). The rate difference observed is attributed to conformational differences among the ions with different charge states.     

Pulsed helium introduction into a quadrupole ion trap for reduced collisional quenching during infrared multiphoton dissociation of electrosprayed ions

Previous infrared multiphoton dissociation (IRMPD) experiments utilizing a quadrupole ion trap mass spectrometer yielded limited photodissociation efficiencies. Helium buffer gas continuously infused into the analyzer region at pressures of typically 1 x 10(-3) Torr to improve ion trap performance can collisionally quench photoexcited ions during the IRMPD process. Photodissociation experiments have indicated that uncorrected pressures below 2 x 10(-5) Torr are necessary to avoid collisional deactivation of photoexcited ions. This paper describes IRMPD in the quadrupole ion trap at reduced pressures utilizing a dual-pulsed introduction of helium buffer gas incorporated into the ion trap scan function. The pulsed introduction of helium buffer gas before ion injection allows the efficient trapping of ions injected from an electrospray source and the removal of helium before laser irradiation. A second pulse of helium directly before ion detection improves the intensity of the ion signal. The use of this dual-pulsed inlet of helium for improved IRMPD is demonstrated with the carbohydrate antibiotics neomycin and erythromycin. Copyright 2000 John Wiley & Sons, Ltd.     

Infrared multiple photon dissociation in the quadrupole ion trap via a multipass optical arrangement

The design of a novel multipass optical arrangement for use with infrared multiple photon dissociation (IRMPD) in the quadrupole ion trap is presented. This design circumvents previous problems of limited IR laser power, small IR absorption cross sections for many molecules, and the limited ion statistics of trapping and detection of ions for IRMPD in the quadrupole ion trap. In contrast to previous designs that utilized the quadrupole ion store, the quadrupole ion trap was operated in the mass selective instability mode with concurrent resonance ejection. The instrumental design consisted of a modified ring electrode with three spherical concave mirrors mounted on the inner surface of the ring. This modified design allowed for eight laser passes across the radial plane of the ring electrode. IRMPD of protonated bis(2-methoxyethyl)ether (diglyme) was used to characterize the performance of the multipass ring electrode. Two consecutive reactions for the IRMPD of protonated diglyme were observed with a lower energy channel predominant at less than 0.6 J (irradiation times from 1 to 30 ms) and a second channel predominant at energies greater than 0.6 J (irradiation times > 30 ms). Other studies presented include a discussion of the dissociation kinetics of protonated diglyme, the use of a pulsed valve for increased trapping efficiency of parent ion populations, and the effects of laser wavelength and of ion residence time in the radial plane of the ring electrode on photodissociation efficiency.     

Multiphoton infrared laser photochemistry of polyatomic molecules: (CF3)3 Cl

Both the primary and resulting yields of the multiphoton dissociation (MPD) of (CF₃)₃Cl upon CO₂-laser irradiation are found to depend strongly on the CO₂-laser radiation frequency. The consecutive multiphoton fragmentation of (CF₃₃C radicals produced in the MPD of (CE₃)₃Cl is shown to occur at several laser frequencies during the same CO₂-laser pulse. The increase in the isotopic selectivity of the overall MPD process due to multiphoton fragmentation and hence the possibility of selective MPD at higher pressures are discussed.     

Application of sustained off-resonance irradiation infrared multiphoton dissociation tandem mass spectrometry to the analysis of biomolecules

A modified pulse sequence for infrared multiphoton dissociation (IRMPD) experiments on a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer in conjunction with sidekick trapping is presented. For IRMPD tandem mass spectrometry experiments gated trapping is normally applied. It ensures that the ions remain on-axis and, thus, cross the laser beam which is aligned on-axis in commercially available instruments. Sidekick trapping is used to capture more ions in the ICR cell in order to increase the signal intensity. However, it may lead to off-axis ion motion, which reduces or even excludes interaction with the laser beam. In this contribution sustained off-resonance irradiation (SORI) was applied to overcome this disadvantage of sidekick trapping. SORI is normally used in conjunction with collision-induced dissociation (CID) experiments to increase the kinetic energy of the ions. Here, SORI is used to influence the cyclotron motion during the laser irradiation time, which leads to temporary intersection of the ion trajectory with the laser beam. With this easy-to-handle experimental setup, IRMPD of ions captured with sidekick trapping leads again to the generation of fragment ions as is demonstrated with several biologically relevant samples like peptides, lipids and glycolipids.