Ion funnel trap interface for orthogonal time-of-flight mass spectrometry

A combined electrodynamic ion funnel and ion trap coupled to an orthogonal acceleration (oa)-time-of-flight mass spectrometer was developed and characterized. The ion trap was incorporated through the use of added terminal electrodynamic ion funnel electrodes enabling control over the axial dc gradient in the trap section. The ion trap operates efficiently at a pressure of approximately 1 Torr, and measurements indicate a maximum charge capacity of approximately 3 x 10(7) charges. An order of magnitude increase in sensitivity was observed in the analysis of low concentration peptides mixtures with orthogonal acceleration (oa)-time-of-flight mass spectrometry (oa-TOF MS) in the trapping mode as compared to the continuous regime. A signal increase in the trapping mode was accompanied by reduction in the chemical background, due to more efficient desolvation of, for example, solvent related clusters. Controlling the ion trap ejection time was found to result in efficient removal of singly charged species and improving signal-to-noise ratio (S/N) for the multiply charged analytes.     

Ion trap/ion mobility/quadrupole/time-of-flight mass spectrometry for peptide mixture analysis

An ion trap/ion mobility/quadrupole/time-of-flight mass spectrometer has been developed for the analysis of peptide mixtures. In this approach, a mixture of peptides is electrosprayed into the gas phase. The mixture of ions that is created is accumulated in an ion trap and periodically injected into a drift tube where ions separate according to differences in gas-phase ion mobilities. Upon exiting the drift tube, ions enter a quadrupole mass filter where a specific mass-to-charge (m/z) ratio can be selected prior to collisional activation in an octopole collision cell. Parent and fragment ions that exit the collision cell are analyzed using a reflectron geometry time-of-flight mass spectrometer. The overall configuration allows different species to be selected according to their mobilities and m/z ratios prior to collision-induced dissociation and final MS analysis. A key parameter in these studies is the pressure of the target gas in the collision cell. Above a critical pressure, the well-defined mobility separation degrades. The approach is demonstrated by examining a mixture of tryptic digest peptides of ubiquitin.     

Characterization of phosphoantigens by high-performance anion-exchange chromatography-electrospray ionization ion trap mass spectrometry and nanoelectrospray ionization ion trap mass spectrometry

New phosphorylated microbial metabolites referred to as phosphoantigens activate immune responses in humans. Although these molecules have leading applications in medical research, no direct method allows their rapid and unambiguous structural identification. Here, we interfaced online HPAEC (high performance anion-exchange chromatography) with ESI-ITMS (electrospray ionization ion trap mass spectrometry) to identify such pyrophosphorylated molecules. A self-regenerating anion suppressor located upstream of electrospray ionization enabled the simultaneous detection of pyrophosphoester by conductimetry, UV and MS. By HPAEC-ITMS and HPAEC-ITMS2, a single run permitted characterization of reference phosphoantigens and of related structures. Although all compounds were resolved by HPAEC, MS enabled their detection and identification by [M-H]- and fragment ions. Isobaric phosphoantigen analogues were also separated by HPAEC and distinguished by MS2. The relevance of this device was demonstrated for phosphoantigens analysis in human urine and plasma. Furthermore, identification of natural phosphoantigens by automatically generated 2D mass spectra from nano-ESI-ITMS is presented. This last technique permits the simultaneous performance of molecular screening of natural phosphoantigen extracts and their identification.     

Application of liquid chromatography/quadrupole-linear Ion trap mass spectrometry and time-of-flight mass spectrometry to the determination of pharmaceuticals and related contaminants in wastewater

This paper describes an enhanced liquid chromatography-mass spectrometry (LC-MS) strategy for the analysis of a selected group of 56 organic pollutants in wastewater. This group comprises 38 pharmaceuticals and 10 of their most frequent metabolites, 6 pesticides, and 2 disinfectants. The LC-MS methodology applied is based in the use of a hybrid triple-quadrupole linear ion trap mass spectrometer (QTRAP) in combination with time-of-flight mass spectrometry (TOF-MS). The join application of both techniques provided very good results in terms of accurate quantification and unequivocal confirmation. Quantification was performed by LC-QTRAP-MS operating under selected reaction monitoring (SRM) mode in both positive and negative electrospray ionization. Unequivocal identification was provided by the acquisition of three SRM transitions per compound in most of the cases and by LC-TOF-MS analysis, which allows obtaining accurate mass measurements of the identified compounds with errors lower than 2 ppm. Additionally, the use of TOF-MS permits retrospective analysis, since the full spectrum is recorded at all times with a high sensitivity. Thus, review of recorded chromatograms looking for new compounds or transformation products suspected to be present in the samples is feasible allowing one to increase the scope of the method along the monitoring program. The analytical performance of the quantitative LC-QTRAP-MS method was evaluated in effluent wastewater samples. Linearity of response over 3 orders of magnitude was demonstrated for most compounds (R(2) > 0.99). Method limits of detection were between 0.04 and 50 ng L(-1). Finally, the methodology was successfully applied to a monitoring study intended to characterize wastewater effluents of six sewage treatment plants in Spain. The presence of most of compounds was detected at concentrations ranging from 9 ng L(-1) (atrazine) to 15 microg L(-1) (paraxanthine).     

Ion probe mass spectrometry: Overview

The bombardment of a surface by 1–25 keV primary ions causes the upper atomic layers to be “sputtered” or stripped off. Most of the material leaves as neutral atoms or molecules, but a small fraction is ejected as positive or negative ions. These secondary ions can then be characterized by a mass spectrometer to provide an analysis of that portion of the material sampled by the primary ion beam. At any one instant, the excitation volume is bounded by the primary ion beam periphery (1–300 μm) and extends to a depth of 5–50 Å depending on experimental conditions. By mass spectrometrically monitoring one or several masses, the analyst can obtain isotopic and concentration in-depth profiles of the sample constituents. Alternatively he can obtain a complete mass spectrum as the primary beam erodes the sample.Control and localization of the primary ion beam erosion permits chemical analysis with lateral x, y resolutions of approximately 1 μm, the examination of fractional surface monolayers, an in-depth analysis with better than 100 Å resolution, and the acquisition of secondary ion images. The mass spectrometer provides elemental coverage from hydrogen to uranium, isotopic characterization and detection limits of 10^-15?10^-19g, depending on the element under consideration.This paper will discuss the production of secondary ions, the various types of instrumentation used, and applications of the ion probe technique.     

Atmospheric pressure MALDI/ion trap mass spectrometry

A new sample ionization technique, atmospheric pressure matrix-assisted laser desorption/ionization (AP MALDI), was coupled with a commercial ion trap mass spectrometer. This configuration enables the application-specific selection of external atmospheric ionization sources: the electrospray/APCI (commercially available) and AP MALDI (built in-house), which can be readily interchanged within minutes. The detection limit of the novel AP MALDI/ion trap is 10-50 fmol of analyte deposited on the target surface for a four-component mixture of peptides with 800-1700 molecular weight. The possibility of peptide structural analysis by MS/MS and MS3 experiments for AP MALDI-generated ions was demonstrated for the first time.     

Broad-band fourier transform quadrupole ion trap mass spectrometry

Broad-band nondestructive ion detection is achieved in a quadrupole ion trap mass spectrometer by impulsive excitation of a collection of trapped ions of different masses and recording of ion image currents induced on a small detector electrode embedded in but isolated from the adjacent end cap electrode. The image currents are directly measured using a simple differential preamplifier, filter, and amplifier combination and then Fourier analyzed to obtain broad-band frequency domain spectra characteristic of the sample ions. The use of the detector electrode provides a significant reduction in capacitive coupling with the ring electrode. This minimizes coupling of the rf drive signal, which can saturate the front-end stage of the detection circuit and prevent measurement of the relatively weaker ion image currents. Although impulsive excitation is preferred due to its broad-band characteristics and simplicity of use, results are also given for narrow-band ac and broad-band SWIFT (stored wave-form inverse Fourier transform) excitation. Data using argon, acetophenone, and n-butylbenzene show that a resolution of better than 1000 is obtained with a detection bandwidth of 400 kHz. An advantage of nondestructive ion detection is the ability to measure a single-ion population multiple times. This is demonstrated using argon as the sample gas with an average remeasurement efficiency of >90%. Tandem mass spectrometry experiments using a population of acetophenone ions are also shown.     

ESI离子阱质谱仪真空系统设计

自主研发质谱仪并能够将其产业化生产是一项十分有意义的工作.质谱仪的工作原理决定了其核心部件(质量分析器和检测器)需要在真空下工作,因此质谱仪的真空系统设计是质谱仪研发工作的基础.本文针对ESI离子阱质谱仪科研样机进行真空系统设计.     

Ion soft landing using a rectilinear ion trap mass spectrometer

A new ion soft landing instrument has been built for the controlled deposition of mass selected polyatomic ions. The instrument has been operated with an electrospray ionization source; its major components are an electrodynamic ion funnel to reduce ion loss, a 90-degree bent square quadrupole that prevents deposition of fast neutral molecules onto the landing surface, and a novel rectilinear ion trap (RIT) mass analyzer. The ion trap is elongated (inner dimensions: 8 mm x 10 mm x 10 cm). Three methods of mass analysis have been implemented. (i) A conventional mass-selective instability scan with radial resonance ejection can provide a complete mass spectrum. (ii) The RIT can also be operated as a continuous rf/dc mass filter for isolation and subsequent soft landing of ions of the desired m/ z value. (iii) The 90-degree bent square quadrupole can also be used as a continuous rf/dc mass filter. The mass resolution (50% definition) of the RIT in the trapping mode (radial ion ejection) is approximately 550. Ions from various test mixtures have been mass selected and collected on fluorinated self-assembled monolayers on gold substrates, as verified by analysis of the surface rinses. Desorption electrospray ionization (DESI) has been used to confirm intact deposition of [Val (5)]-Angiotensin I on a surface. Nonmass selective currents up to 1.1 nA and mass-selected currents of up to 500 pA have been collected at the landing surface using continuous rf/dc filtering with the RIT. A quantitative analysis of rinsed surfaces showed that the overall solution-to-solution soft landing yields are between 0.2 and 0.4%. Similar experiments were performed with rf/dc isolation of both arginine and lysine from a mixture using the bent square quadrupole in the rf/dc mode. The unconventional continuous mass selection methods maximize soft landing yields, while still allowing the simple acquisition of full mass spectra.     

Ultraviolet photodissociation at 355 nm of fluorescently labeled oligosaccharides

Ultraviolet photodissociation (UVPD) produces complementary fragmentation to collision-induced dissociation (CID) when implemented for activation of fluorescently labeled oligosaccharide and glycan ions. Reductive amination of oligosaccharides with fluorophore reagents results in efficient photon absorption at 355 nm, producing fragment ions from the nonreducing end that do not contain the appended fluorophore. In contrast to the fragment ions observed upon UVPD (A- and C-type ions), CID produces mainly reducing end fragments retaining the fluorophore (Y-type ions). UVPD affords better isomeric differentiation of both the lacto-N-fucopentaoses series and the lacto-N-difucohexaoses series, but in general, the combination of UVPD and CID offers the most diagnostic elucidation of complex branched oligosaccharides. Four fluorophores yielded similar MS/MS results; however, 6-aminoquinoline (6-AQ), 2-amino-9(10 H)-acridone (AMAC) and 7-aminomethylcoumarin (AMC) afforded more efficient photon absorption and subsequent dissociation than 2-aminobenzamide (2-AB). UVPD also was useful for characterization of glycans released from ribonuclease B and derivatized with 6-AQ. Lastly, electron photodetachment dissociation of oligosaccharides derivatized with 7-amino-1,3-naphthalenedisulfonic acid (AGA) yielded unique cross-ring cleavages similar to those obtained by electron detachment dissociation.     

Ion mobility mass spectrometry analysis of human glycourinome

Complex carbohydrates are macromolecules biosynthesized in nontemplate-type processes, bearing specific glycoepitopes involved in crucial recognition processes such as cell differentiation and cell-cell interactions. Chemical structure of single components in complex mixtures can be analyzed by mass spectrometry for determination of the size and sequence of monosaccharides involved, branching patterns, and substitution by fucose and sialic acids. For de novo identification of glycoforms in human urinome containing N- and O-free and amino acid-linked oligosaccharides, a novel method of ion mobility tandem mass spectrometry followed by computer-assisted assignment is described. Distinct patterns of ions nested specifically by their m/z values and their drift time are observed by IMS-MS. An additional peak capacity for identification of time-separated m/z values in the IMS TOF MS mode for differentiation of singly, doubly, and triply charged molecular ion species by ion mobility separation contributes to significant reduction of carbohydrate complexity in a given mass window. Profiling of glycoforms from human urinome represents a highly efficient approach for biomarker discovery and differential glycotarget identification, demonstrating potential for diagnosis of human diseases, as for congenital disorders of glycosylation.     

High-speed mass analysis of whole erythrocytes by charge-detection quadrupole ion trap mass spectrometry

Herein, we report an application of charge-detection quadrupole ion trap mass spectrometry to the measurement of total dry masses of mammalian and poultry erythrocytes evaporated/ionized by laser-induced acoustic desorption. The method is rapid and widely applicable. Eight different types of red blood cells (RBCs) have been successfully analyzed, including those of human, goat, cow, mouse, pig, and chicken. The measured mean masses (weights per corpuscle) range from 0.58 x 10(13) Da (9.6 pg) of goat RBCs to 2.80 x 10(13) Da (46.5 pg) of chicken RBCs. The total dry weights determined for human RBCs from a healthy male adult, a patient with iron-deficiency anemia, and a patient with thalassemia are 34.8, 28.8, and 20.6 pg, respectively. These weights, except that of thalassemia, are all approximately 10% higher than their corresponding mean corpuscular hemoglobin values determined by a commercial automated hematology analyzer. The mass distribution profiles of the cells are all near-Gaussian, with a standard deviation of 15% for the normal human RBCs. The deviation increases significantly to 20% for RBCs with thalassemia characteristics and 27% for RBCs with iron-deficiency anemia characteristics. All the observations are in accord with their corresponding mean corpuscular volume measurements, indicating an increase in anisocytosis (variation in RBC size) in the anemic samples. Our results suggest a broad and promising application of this new technology to high-speed mass analysis of RBCs and other biological whole cells as well.     

Ultra-stable nanoparticles of CdSe revealed from mass spectrometry

Nanoparticles under a few nanometres in size have structures and material functions that differ from the bulk because of their distinct geometrical shapes and strong quantum confinement. These qualities could lead to unique device applications. Our mass spectral analysis of CdSe nanoparticles reveals that (CdSe)(33) and (CdSe)(34) are extremely stable: with a simple solution method, they grow in preference to any other chemical compositions to produce macroscopic quantities. First-principles calculations predict that these are puckered (CdSe)(28)-cages, with four- and six-membered rings based on the highly symmetric octahedral analogues of fullerenes, accommodating either (CdSe)(5) or (CdSe)(6) inside to form a three-dimensional network with essentially heteropolar sp(3)-bonding. This is in accordance with our X-ray and optical analyses. We have found similar mass spectra and atomic structures in CdS, CdTe, ZnS and ZnSe, demonstrating that mass-specified and macroscopically produced nanoparticles, which have been practically limited so far to elemental carbon, can now be extended to a vast variety of compound systems.     

Collisional activation with random noise in ion trap mass spectrometry.

Random noise applied to the end caps of a quadrupole ion trap is shown to be an effective means for the collisional activation of trapped ions independent of mass/charge ratio and number of ions. This technique is compared and contrasted with conventional single-frequency collisional activation for the molecular ion of N,N-dimethylaniline, protonated cocaine, the molecular anion of 2,4,6-trinitrotoluene, and doubly pronated neuromedin U-8. Collisional activation with noise tends to produce more extensive fragmentation than the conventional approach due to the fact that product ions are also kinetically excited in the noise experiment. The efficiency of the noise experiment in producing detectable product ions relative to the conventional approach ranges from being equivalent to being a factor of 3 less efficient. Furthermore, discrimination against low mass/charge product ions is apparent in the data from multiply charged biomolecules. Nevertheless, collisional activation with random noise provides a very simple means for overcoming problems associated with the dependence of single-frequency collisional activation on mass/charge ratio and the number of ions in the ion trap.     

Two ions in one trap: ultra-high precision mass spectrometry?

By trapping two different ions in the same Penning trap at the same time, we simultaneously measured the ratio of their cyclotron frequencies (from which we obtain their atomic mass ratio) with a precision of about 10/sup -11/ in only a few hours. In order to perform these comparisons, we must be able to measure and control all three normal modes of motion of each ion - cyclotron, axial, and magnetron. We have developed novel techniques to do, so and we are currently using these tools to carefully investigate the important question of systematic errors in those measurements. This new technique shows promise to expand the precision of mass spectrometry an order of magnitude beyond the current state-of-the-art.     

Ion source with magnetic field for time-of-flight mass spectrometry

A new ion source for the time-of-flight mass spectrometer (TOFMS) with electron-impact ionization and built-in permanent magnet for electron-beam focusing is described. The focusing of electrons in a homogeneous field leads to an increase in the resolving power and sensitivity of the TOFMS. A spectrometer with the proposed source can operate at electron energies within the 7–100-eV range. The method of suppressing spurious lines in the mass spectrum is described, which is based on a decrease in the energy of ionizing electrons.     

Fourier transform time-of-flight mass spectrometry in an electrostatic ion beam trap

We report on the application of an electrostatic ion beam trap as a mass spectrometer. The instrument is analogous to an optical resonator; ions are trapped between focusing mirrors. The storage time is limited by the residual gas pressure and reaches up to several seconds, resulting in long ion flight paths. The oscillation of ion bunches between the mirrors is monitored by nondestructive image charge detection in a field-free region and mass spectra are obtained via Fourier transform. The principle of operation is demonstrated by measuring the mass spectrum of trapped Ar+ and Xe+ particles, produced by a standard electron impact ion source. Also, mass spectra of heavier PEGnNa+ and bradykinin ions from a pulsed MALDI ion source were obtained. The long ion flight path, combined with mass-independent charge detection, makes this system particularly interesting for the investigation of large molecules.     

Automated gain control ion funnel trap for orthogonal time-of-flight mass spectrometry

Time-of-flight mass spectrometry (TOF MS) is increasingly used in proteomics research. Herein, we report on the development and characterization of a TOF MS instrument with improved sensitivity equipped with an electrodynamic ion funnel trap (IFT) that employs an automated gain control (AGC) capability. The IFT-TOF MS was coupled to a reversed-phase capillary liquid chromatography (RPLC) separation and evaluated in experiments with complex proteolytic digests. When applied to a global tryptic digest of Shewanella oneidensis proteins, an order-of-magnitude increase in sensitivity compared to that of the conventional continuous mode of operation was achieved due to efficient ion accumulation prior to TOF MS analysis. As a result of this sensitivity improvement and related improvement in mass measurement accuracy, the number of unique peptides identified in the AGC-IFT mode was 5-fold greater than that obtained in the continuous mode.