High-sensitivity capillary electrophoresis-electrospray ionization quadrupole ion trap time-of-flight mass spectrometry (CE-ESI-QIT-TOF MS) was developed to structurally characterize four kinds of pyridylaminated (PA) oligosaccharides, i.e., lactose (Lac)-PA, globotriose (Gb3)-PA, globotetraose (Gb4)-PA, and IV(3) αGalNAc-Gb4 (Forssman antigen)-PA, derived from neutral glycosphingolipids. The CE-MS system included the head-column field-amplified sample stacking (HC-FASS) method for effective sample injection into a capillary column in CE, a sheathless interface between CE and a mass spectrometer, and MS and tandem MS (MS(2)) measurements with narrow mass range repeated high-speed switching. The total sensitivity of the developed CE-MS system was about 20,000 times higher than that of the conventional CE-MS system consisting of pressure injection, a sheath-flow interface, and a wide mass range measurement. The MS and MS(2) spectra of the four PA-oligosaccharides at a concentration of 25 amol/μL in mixtures (each 250 amol/10 μL in a tube) clearly showed protonated molecular ions ([M + H](+)) and the fragment ions responsible for the sequential elimination of saccharides. The developed CE-MS system is a powerful method for the structural characterization of glycosphingolipids extracted from very small amounts of biological materials and could be extended to the structural characterization of oligosaccharides derived from glycoproteins.
2784 Letter to the Editor range, which may not be acceptable for quantitative analysis. Compared with PDA detection, a 10-fold decrease in LOQs was obtained by APPI-MS or APCI-MS.APPI-MS was tested for analysis of fullerenes. The preliminary experiment for trace analysis of fullerenes by LC/APPI-MS was examined. The overall performance of APPI-MS was comparable to that of APCI-MS for determination of C 60 and C 70 . The method would be applicable to quan-titative analysis of fullerenes in environmental or biological samples.
The atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) is a quite convenient soft ionization for biomolecules, keeping analytes atmospheric conditions instead of high vacuum conditions. In this study, an AP-MALDI ion source has been coupled to a quadrupole ion trap time-of-flight (QIT-TOF) mass spectrometer, which is able to perform MS(n) analysis. We applied this system to the structural characterization of monosialogangliosides, GM1 (NeuAc) and GM2 (NeuAc), disialogangliosides, GD2 (NeuAc, NeuAc), GD1a (NeuAc, NeuAc) and GD1b (NeuAc, NeuAc) and trisialoganglioside GT1a (NeuAc, NeuAc, NeuAc). In this system, the negative ion mass spectra of MS, MS(2) and MS(3), a set of three mass spectra, were able to measure within 2 s per cycle. Thus, obtained results demonstrate that the negative ion mode MS, MS(2) and MS(3) spectra provided sufficient information for the determination of molecular weights, oligosaccharide sequences and ceramide structures, and indicate that the AP-MALDI-QIT-TOF mass spectrometry keeping analytes atmospheric conditions with MS(n) switching is quite useful and convenient for structural analyses of various types of sialic acid-containing GSLs, gangliosides.
An ion optical design of a high resolution multi-turn time-of-flight mass analyzer (MT-TOF MA) is presented. The analyzer has rotationally symmetric main electrodes with additional mirror symmetry about a mid-plane orthogonal to the axis of symmetry. Rotational symmetry allows a higher density of turns in the azimuthal (drift) direction compared to MT-TOF MAs that are linearly extended in the drift direction. Mirror symmetry about a mid-plane helps to achieve a high spatial isochronicity of the ions’ motion. The analyzer comprises a pair of polar-toroidal sectors S1 and S3, a pair of polar (trans-axial) lenses, and a pair of conical lenses for longitudinal and lateral focusing. A toroidal sector S2 located at the mid-plane of the analyzer has a set of embedded drift focusing segments providing focusing and spatial isochronicity in the drift direction. The ions’ drift in the azimuthal direction can be reversed by using dedicated reversing deflectors. This gives the possibility of several operational modes with different numbers of turns and passes in the drift direction. According to numerical simulations, the mass resolving power of the analyzer ranges from [Formula: see text]40 k (fwhm) at small (typically below ten) numbers of turns to [Formula: see text]450 k (fwhm) at 96 turns.
Four types of neutral glycosphingolipids (LacCer, Gb3Cer, Gb4Cer, and IV3αGalNAc-Gb4Cer; 10 pmol each) were analyzed using high-performance liquid chromatography (HPLC)-electrospray ionization quadrupole ion trap time-of-flight (ESI-QIT-TOF) mass spectrometry (MS) with a repeated high-speed polarity and MSn switching system. This system can provide six types of mass spectra, including positive and negative ion MS, MS2, and MS3 spectra, within 1 s per cycle. Using HPLC with a normal-phase column, information on the molecular weights of major molecular species of four neutral glycosphingolipids was obtained by detecting [M+Na]+ in the positive ion mode mass spectra and [M−H]− in the negative ion mode mass spectra. Sequences of glycosphingolipid oligosaccharide were obtained in the negative ion MS2 spectra. In addition, information on the ceramide structures was clearly obtained in the negative ion MS3 mass spectra. GlcCer molecular species were analyzed by HPLC-ESI-QIT-TOF MS with a reversed-phase column using 1 pmole of GlcCer. The structures of the seven molecular species of GlcCer, namely, d18:1-C16:0, d18:1-C18:0, d18:1-C20:0, d18:1-C22:0, d18:1-C23:0, d18:1-C24:1, and d18:1-C24:0, were characterized using positive ion MS and negative ion MS, MS2, and MS3. The established HPLC-ESI-QIT-TOF MS with MSn switching and a normal phase column has been successfully applied to the structural characterization of LacCer and Gb4Cer in a crude mixture prepared from human erythrocytes.
Orbital frequency
analyzer (OFA) is one of the electrostatic ion
trap mass analysers for FTMS, which offers ultrahigh mass resolution
and high ion charge capacity. In order to analyze multiply charged
proteins and other large biological particles by means of charge detection
mass spectrometry, a data processing algorithm was created to suit
the image charge signal of nonharmonic waveform nature. The algorithm
is capable of detecting collisions between ions and residual gas molecules,
to determine lifetime of ions, and to evaluate the charge and mass
values for ions having lifetime above a threshold. With the filtering
of the lifetime and charge value, the chemical noise from small molecules
and protein fragments can be eliminated in the reconstructed spectrum,
facilitating measurement of protein content at a very low concentration,
down to tens of nanomolars. The standard deviation of charge measurement
is between 1.1 to 1.8 e for ions with oscillation lifetimes from 1
to 0.4 s, and this in turn determines the CDMS spectrum mass peak
width. It has been found that the lower voltage setting of the OFA
results in a larger population of ions surviving for longer times
and thus produces narrower mass peak width. While OFA is able to run
multiplexed CDMS without ion motion interference, coexisting ions
of the same or very close m/z can
cause interference between their image charge signals, which increases
the error in charge determination.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.