Infrared multiple photon dissociation spectrum of protonated bis(2-methoxyethyl) ether obtained with a tunable CO2 laser.

A moderate-resolution infrared multiple photon dissociation (IRMPD) spectrum of protonated bis(2-methoxyethyl) ether (diglyme) was obtained using a grating-tuned CO2 laser. The experimental spectrum compares well with one calculated theoretically at the MP2 level and exhibits defined peaks over the span of the CO2 laser output lines as opposed to a relatively featureless spectrum over this wavelength range obtained using free electron laser infrared radiation. The lowest energy structure corresponding to the calculated vibrational spectrum is consistent with structures previously calculated at the same level of theory. Alternative structures were calculated at lower levels of theory for comparison and investigation of the energetics of proton-heteroatom interactions. Broadening of the IRMPD action spectrum due to energetic phenomena characteristic of proton bridges was not observed and thus did not obscure the correlation between theoretical calculations and experimentally determined spectra as it may have in previous studies.

Bioconjugation of ribonuclease A: a detailed chromatographic and mass spectrometric analysis of chemical modification by a cross-linking reagent.

The modification of ribonuclease A with the heterobifunctional cross-linker, 4-succinimdyloxycarbonyl-methyl-alpha-[2-pyridyldithio]-toluene (SMPT) is described. RNase A has 11 potential sites of modification by the SMPT reagent. Tracking the two-dimensional separation and proteolytic digestion of SMPT-modified RNase A with ESI/FTICR-MS and HPLC/ESI/QIT-MS demonstrates the detailed information about number of SMPT modifications and sites of modification that can be obtained by application of these techniques. Analysis of native and modified RNase A tryptic digests by ESI/FTICR-MS resulted in the identification of the sites of modification. Semiquantitative results of the reactivity of certain lysine residues toward the coupling reagent SMPT are presented. Two sites (lysines 1 and 37) are highly reactive, while three sites (lysines 41, 61, and 104) appear to be unreactive toward SMPT under the conditions used. Experimental results demonstrate that quantitative comparison of relative intensities of peptide sequences of different charge states is not possible. No correlation was found between number of basic residues and sensitivity to detection. Digestion of the modified and unmodified RNase A by subtilisin followed by examination by HPLC/ESI/QIT-MS and MS(n) enabled further investigation of modification on lysines 1 and 7, including modification at the epsilon- and alpha-amino positions on lysine 1.

Assessing enzyme substrate specificity using combinatorial libraries and electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry.

A model experiment for the 'on-line' screening of substrate libraries by enzymes using combinatorial libraries in combination with electrospray ionization-Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry has been performed. The reaction between the electrophilic substrate 1-chloro-2,4-dinitrobenzene and component of a H-gamma-Glu-Cys-Xxx-OH library, catalyzed by glutathione-S-transferase, has been monitored. It shows the feasibility of 'two-dimensional' screening of substrate libraries by ESI-FTICR mass spectrometry.

Analysis of combinatorial libraries using electrospray Fourier transform ion cyclotron resonance mass spectrometry.

Electrospray ionization coupled with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry has been used to provide information about complete combinatorial libraries of small peptides containing 10(3)-10(4) components. The fidelity of attempted synthesis steps can be ascertained rapidly, and, when the extremely high resolution FTICR mass spectra are combined with appropriate computer simulation, both diversity and degeneracy of the libraries as synthesized can be assessed.

Pulsed-gas glow discharge for ultrahigh mass resolution measurements with fourier transform ion cyclotron resonance mass spectrometry.

A new pulsed-gas glow discharge (GD) source has been developed for use with an external ion source Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. With pulsed argon gas introduction into the GD source, the gas load and pressure in the mass analyzer region were greatly reduced; this resulted in improved mass resolution. Mass resolution of greater than 1 450 000 (fwhm) has been achieved for Cu(+) ions from a brass sample, the highest reported for any type of GD mass spectrometer. The pulsed-gas GD source promises analytical usefulness for ultrahigh resolution measurements in GD mass spectrometry.

Internal glow discharge-fourier transform ion cyclotron resonance mass spectrometry.

A glow discharge (GD) ion source has been developed to work within the high magnetic field of a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. Characterization of this source revealed that the optimum operating voltage, pressure, and current are significantly lower than those for normal glow discharges. The sputter rate was lowered to 1/30th of that found with a normal glow discharge source operated external to the high magnetic field region. Operation of the GD source closer to the FTICR analyzer cell than with previous experimental designs resulted in improved ion transport efficiency. Preliminary results from this internal GD source have established detection limits in the low parts per million range for selected elemental species.

The mass-to-charge ratio scale.

Increases in the capacity for accurately measuring the mass-to-charge ratio of specific gas-phase ions justify the reconsideration and standard definition of the gas-phase mass-to-charge ratio scale and the clearly denned connection of that scale to condensed phases. We propose that the chemical mass standard for solids and the gas phase be based upon the mass of carbon-12 buckminsterfulierene ((12)C60). The mass-to-charge ratio scale in the gas phase would be based upon the mass of gas-phase (12)C60, the mass of the electron, and the electron charge in atomic units. As mass measurement accuracy improves, corrections to this mass-to-charge ratio standard are anticipated for the vaporization energy of the 12C60 molecule and its ionization potential or electron affinity. We propose that the positive ion scale be set by the mass-to-charge ratio of (12)C 60 (+) as (+)719.9994514±0.0000004 u per electron charge. We propose that the negative ion mass scale be set by the mass-to-charge ratio of (12)C 60 (-) as (-)720.0005484±0.0000004 u per electron charge.

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.

An improved ion guide for external ion injection in glow discharge—fourier transform ion cyclotron resonance mass spectrometry.

To improve the existing ion transport optics of our glow discharge (GD)-Fourier transformion cyclotron resonance (FT-ICR) mass spectrometer, we simulated several ion trajectories between the GD source region and the ICR analyzer cell. These calculations suggested that a number of simple improvements, including the use of an ion flight tube and an electrically isolated conductance limit, would increase the efficiency of ion transfer through the fringing fields of the FT-ICR superconducting magnet and into the ICR analyzer cell. Ion beam intensity was monitored as a function of the distance between the GD source and the analyzer cell before and after implementing these improvements. A twentyfold improvement in the transport efficiency, as well as a fifteenfold enhancement in detected ET-ICR signals, was observed.

Probing trapped ion energies via ion-molecule reaction kinetics: Quadrupole ion trap mass spectrometry.

We present a detailed study of the energies of the ions stored in a quadrupole ion trap mass spectrometer (QITMS). Previous studies have shown that the rate constant, k, for the charge exchange reaction Ar(+) N(+) 2 →, N(+) 2+Ar increases with increasing ion-molecule center-of-mass kinetic energy (K.E.cm). Thus, we have determined k for this chemical "thermometer" reaction at a variety of Ar and N2 pressures and have assigned K.E.cm values as a function of the q2 of the Ar(+) ion both with and without He buffer gas present in the trap. The K.E.cm energies are found to lie within the range 0.11-0.34 eV over the variety of experimental conditions investigated. Quantitative "cooling" effects due to the presence of He buffer gas are reported, as are increases in K.E.cm due to an increase in the q2 of the Ar(+) ion. "Effective" temperatures of the Ar(+) ions in He buffer are determined based on a Maxwell-Boltzmann distribution of ion energies. The resulting temperatures are found to lie within the range ≈ 1700-3300 K. We have also examined the K.E.cm, values arising from the chemical thermometer reaction of O(+) 2 with CH4, as previous assignments of effective ion temperatures based on this reaction have been called into question.

Probing trapped ion energies via ion-molecule reaction kinetics: Fourier transform ion cyclotron resonance mass spectrometry.

The kinetic energy-dependent Ar(+)+ N2 ion-molecule reaction has been used as a chemical "thermometer" to determine the kinetic energy of ions produced by electron ionization and trapped by using a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. The rate constant for this reaction obtained on the FTICR mass spectrometer was compared to previous work, which allowed a kinetic energy estimate to be made. In addition, the effects of varying parameters such as trapping voltage and pressure on ion kinetic energy were investigated. No evidence of the differing reactivity of higher energy electronic states of Ar(+), such as (2)P1/2, was observed and the results of a model of this system are presented that support this observation. Pressure studies revealed that with an average of as few as 13 ion-molecule collisions, Ar(+) ions are collisionally relaxed to an extent unaffected by additional collisions. Based on recent variable temperature selected ion flow drift tube measurements, FTICR ion energies are estimated to be slightly above thermal.

A glow discharge ion source with fourier transform ion cyclotron resonance mass spectrometric detection.

A glow discharge (CD) ion source has been coupled to a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer using a four-element electrostatic lens to accelerate and focus ions generated external to the instrument's high magnetic field into its analyzer cell. Like other CD mass spectrometers, GD-FT-ICR can provide a quantitative measure of bulk analyte concentration with good precision and accuracy. Although detection limits currently attainable are several orders of magnitude higher than the commercially available magnetic sector-based instrument, CD-FT-ICR holds promise for ultrahigh resolving power elemental mass analysis. Several schemes are proposed to lower the detection limits of the technique while still providing high enough resolution to resolve isobaric interferences.

Digital, synchronously timed control system for pulsed ion cyclotron resonance mass spectrometry.

An integrated control system for a pulsed ion cyclotron resonance (ICR) mass spectrometer has been designed, constructed, and put into operation. It utilizes a crystal clock oscillator to accurately control the delay and width of the various pulses necessary for pulsed ICR operation. A switched integrator, sample/hold postdetection system is also described which improves aquisition, display, and signal-to-noise characteristics of the detected ion signal obtained from a gated marginal oscillator detector.