Analysis of cancer cell lipids using matrix-assisted laser desorption/ionization 15-T Fourier transform ion cyclotron resonance mass spectrometry

A combination of methodologies using the extremely high mass accuracy and resolution of 15-T Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry (MS) was introduced for the identification of intact cancer cell phospholipids. Lipids from a malignant glioma cell line were initially analyzed at a resolution of >200,000 and identified by setting the mass tolerance to ±1 mDa using matrix-assisted laser desorption/ionization (MALDI) 15-T FT-ICR MS in positive ion mode. In most cases, a database search of potential lipid candidates using the exact masses of the lipids yielded only one possible chemical composition. Extremely high mass accuracy (<0.1?ppm) was then attained by using previously identified lipids as internal standards. This, combined with an extremely high resolution (>800,000), yielded well-resolved isotopic fine structures allowing for the identification of lipids by MALDI 15-T FT-ICR MS without using tandem mass spectrometric (MS/MS) analysis. Using this method, a total of 38 unique lipids were successfully identified.     

Dipolar and quadrupolar detection using an FT-ICR MS setup at the MPIK Heidelberg

Dipolar and single-phase two-electrode quadrupolar detection schemes have been investigated at a Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) setup built for the KATRIN experiment at the Max-Planck-Institute for Nuclear Physics (MPIK) in Heidelberg. We present first experimental results of ⁷Li^?+? signals from a cylindrical Penning trap configuration for both detection schemes. While the prominent signal of the conventional dipolar detection scheme marks the reduced cyclotron frequency, the main signal for the quadrupolar detection appears at the sum of the reduced cyclotron frequency and the magnetron frequency. For ideal trapping fields, this sum frequency equals the ion cyclotron frequency ?? _c ?=?qB/(2??m). Sidebands due to the combined motions of the cyclotron mode and magnetron mode are observed by quadrupolar detection which allows the determination of the respective combinations of eigenfrequencies.     

Fundamental aspects of FT-ICR and applications to chemistry

Fourier transform ion cyclotron resonance (FT-ICR) spectroscopy, a modern form of mass spectrometry whose advantages were first demonstrated in our laboratory in 1974, is characterized by ultrahigh mass resolution, wide mass range, high speed and automatic mass calibration. Together with the FT-ICR double resonance experiment, these advantages make FT-ICR a powerful technique for studying complex ion/molecule reaction pathways and for general problems in analytical mass spectrometry. In addition, the high resolution principles of FT-ICR have been widely adopted around the world for precise mass measurements of nuclides.     

Depth distribution of SiD and SiD2 complexes in silicon bombarded by 10 keV D+ 2 ion

Abstract

A secondary ion mass spectrometry (SIMS) and a neutral molecule mass spectrometry (NMMS) study of deuterium trapping in a single crystalline silicon as a result of ion irradiation at fluences 10^16--10¹⁸ cm-² are presented. An attempt has been made to observe the formation and evolution of defect profiles containing one or two deuterium atoms (SiD- and SiD₂-complexes). The proposed model of radiation-induced sequential reactions describes satisfactorily the accumulation of SiD- and SiD₂-complexes and the reemission of D₂-molecules.     

Corroding the chromium cation

Two [Cr,O₂]⁺ isomers have been selectively produced and studied by FT-ICR mass spectrometry. The Cr(O₂)⁺ complex was formed by supersonically expanding a plasma produced by laser vaporization of chromium metal with the helium carrier gas, which was seeded with traces of oxygen, while the chromyl cation is formed in an expansion with N₂O. The complex is stable against thermal collisions, but in a bimolecular reaction with water it is rapidly converted to the chromyl cation, with ligand exchange being only a minor side reaction. Isotopic studies suggest a side-on geometry for Cr(O₂)⁺, in accordance with density functional (B3LYP) calculations. The present work indicates that an investigation of the selected isomers can indeed be carried out, if appropriate chemical methods for the ion generation are applied.     

Mass Spectrometry,Review of the Basics: Electrospray,MALDI, and Commonly Used Mass Analyzers

Abstract

Mass spectrometry (MS) has progressed to become a powerful analytical tool for both quantitative and qualitative applications. The first mass spectrometer was constructed in 1912 and since then it has developed from only analyzing small inorganic molecules to biological macromolecules, practically with no mass limitations. Proteomics research, in particular, increasingly depends on MS technologies. The ability of mass spectrometry analyzing proteins and other biological extracts is due to the advances gained through the development of soft ionization techniques such as electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) that can transform biomolecules into ions. ESI can efficiently be interfaced with separation techniques enhancing its role in the life and health sciences. MALDI, however, has the advantage of producing singly charges ions of peptides and proteins, minimizing spectral complexity. Regardless of the ionization source, the sensitivity of a mass spectrometer is related to the mass analyzer where ion separation occurs. Both quadrupole and time of flight (ToF) mass analyzers are commonly used and they can be configured together as QToF tandem mass spectrometric instruments. Tandem mass spectrometry (MS/MS), as the name indicates, is the result of performing two or more sequential separations of ions usually coupling two or more mass analyzers. Coupling a quadrupole and time of flight resulted in the production of high-resolution mass spectrometers (i.e., Q-ToF). This article will historically introduce mass spectrometry and summarizes the advantages and disadvantages of ESI and MALDI along with quadrupole and ToF mass analyzers, including the technical marriage between the two analyzers. This article is educational in nature and intended for graduate students and senior biochemistry students as well as chemists and biochemists who are not familiar with mass spectrometry and would like to learn the basics; it is not intended for mass spectrometry experts.     

Hydrogenated carbon clusters produced by highly charged ion impact on solid

The emission of small (hydrogenated) carbon cluster ions C_nHm ⁺ (n =2-22) upon highly charged Xe^q+ (q =20-44) impact on C₈₄ surfaces is studied by means of time-of-flight secondary ion mass spectrometry. The respective stage of hydrogenation/protonation of a certain carbon cluster ion Cn ⁺ is a strong indication for its geometrical structure. From the cluster ion yield as a function of cluster size it can be concluded, that the hydrogenation takes place after the initial carbon cluster formation. The carbon clusters seem to be emitted as an entity in agreement with “equilibrium” and “shock wave” models. Received 4 February 2000     

Gas‐phase studies of copper catalyzed aerobic cross coupling of thiol esters and arylboronic acids

Electrospray ionization/mass spectrometry (ESI‐MS) is used to monitor a Cu‐catalyzed aerobic cross‐coupling reaction between thiol esters and arylboronic acids. The ESI spectra show the formation of Cu‐complexes with the starting thiol ester 1 and the coupling product. The formation of an ionic complex at m/z 305 is observed, most likely occurring upon the elimination of a mixed anhydride from [(1)CuOAc]⁺. An online monitoring of the reaction using ESI‐MS was carried out allowing calculation of rate constants and thermodynamic parameters (ΔH^?, ΔS^?, and ?G^?) for the title reaction.     

Synthesis, characterization, DNA binding and cleavage studies of mixed-ligand Cu(II) complexes of 2,6-bis(benzimidazol-2-yl)pyridine

Four novel copper(II) complexes of the composition [CuLX] where L = 2,6-bis(benzimidazole-2yl)pyridine, X = dipyridophenazine (L₁), 1,10-phenanthroline (L₂), hydroxyproline (L₃) and 2,6-pyridine dicarboxylic acid (L₄) were synthesized and characterized by using elemental analysis, FT-IR, UV–vis, ESI-MS, molar conductance and magnetic susceptibility measurements. The complexes [CuLL₁](NO₃)₂ [1], [CuLL₂](NO₃)₂ [2], [CuLL₃](NO₃) [3] and [CuLL₄] (NO₃) [4] are stable at room temperature. In DMSO the complexes [1] and [2] are 1:2 electrolytes, [3] and [4] are 1:1 electrolytes. Based on elemental and spectral studies five coordinated geometry is assigned to all the four complexes. The interaction of four copper ion complexes with calf thymus DNA were carried out by UV–vis titrations, fluorescence spectroscopy, thermal melting and viscosity measurements .The binding constant (K_b) of the above four metal complexes were determined as 5.43 × 10⁴ M_,⁻¹ 2.56 × 10⁴ M⁻¹, 1.21 × 10⁴ M⁻¹ and 1.57 × 10⁴ M⁻¹ respectively. Quenching studies of the four complexes indicates that these complexes strongly bind to DNA, out of all complex 1 is binding more strongly. Viscosity measurements indicate the binding mode of complexes with CT DNA by intercalation through groove. Thermal melting studies also support intercalative binding. The nuclease activity of the above metal complexes shows that 1, 2 and 3 complexes cleave DNA through redox chemistry.     

Trace Analysis Using Position-Sensitive Detection in Magnetic Sector Mass Spectrometry

Abstract

A position-sensitive ion detection system for trace analysis with magnetic sector mass spectrometers is described in detail, with particular application to high temperature mass spectrometry. The detection system consists of two stacked microchannel plates (Chevron assembly) backed by a resistive anode encoder and associated electronics. The range of masses simultaneously detectable is m to 1.2m. For electron impact ionization of silver at an electron energy of 10.5 eV, the sensitivity is 1.6 × 10^?7 Pa, and the mass resolution is 260 at mass 80 (valley 10% of the peak height definition). Additional applications for the detection system are discussed.     

Isotope shifts and hyperfine structure in the transitions of gadolinium

High-resolution resonance ionization mass spectrometry has been used to measure isotope shifts and hyperfine structure in all (J = 2-6) and the transitions of gadolinium (Gd I). Gadolinium atoms in an atomic beam were excited with a tunable single-frequency laser in the wavelength range of 422-429 nm. Resonant excitation was followed by photoionization with the 363.8 nm line of an argon ion laser and resulting ions were mass separated and detected with a quadrupole mass spectrometer. Isotope shifts for all stable gadolinium isotopes in these transitions have been measured for the first time. Additionally, the hyperfine structure constants of the upper states have been derived for the isotopes ^{155, 157} Gd and are compared with previous work. Using prior experimental values for the mean nuclear charge radii, derived from the combination of muonic atoms and electron scattering data, field shift and specific mass shift coefficients for the investigated transitions have been determined and nuclear charge parameters for the minor isotopes ^{152, 154} Gd have been calculated. Received 18 November 1999     

Maternal obesity does not influence human milk protein 15N natural isotope abundance

ABSTRACT

Obesity increases protein metabolism with a potential effect on nitrogen isotope fractionation. The aim of this study was to test the influence of obesity on human milk extracted protein ¹⁵N natural isotope abundance (NIA) at one month post-partum and to compare human milk extracted protein ¹⁵N NIA and bulk infant hair ¹⁵N NIA. This cross-sectional observational study involved 16 obese mothers (body mass index (BMI)?≥?30?kg?m^?2 before pregnancy) matched with 16 normal-weight mothers (18.5?kg?m^?2?≤?BMI?<?25?kg?m^?2) for age and pregnancy characteristics. Human milk extracted protein and bulk infant hair ¹⁵N NIA were determined by isotope ratio monitoring by mass spectrometry interfaced to an elemental analyser (IRM-EA/MS). No significant difference was found in human milk protein ¹⁵N NIA values between obese and normal-weight mothers (8.93?±?0.48?‰ vs. 8.95?±?0.27?‰). However, human milk protein ¹⁵N NIA was significantly lower than bulk infant hair ¹⁵N NIA: 8.94?±?0.38?‰ vs. 9.66?±?0.69?‰, respectively. On the basis of these results, it is concluded that human milk protein ¹⁵N NIA measured at one month post-partum is not influenced by maternal obesity. These findings suggest that ¹⁵N NIA may be exploited to study metabolism without considering maternal obesity as a confounder.     

Recent developments in ion detection techniques for Penning trap mass spectrometry at TRIGA-TRAP

The highest precision in the determination of nuclear and atomic masses can be achieved by Penning trap mass spectrometry. The mass value is obtained through a measurement of the cyclotron frequency of the stored charged particle. Two different approaches are used at the Penning trap mass spectrometer TRIGA-TRAP for the mass determination: the destructive Time-Of-Flight Ion Cyclotron Resonance (TOF-ICR) technique and the non-destructive Fourier Transform Ion Cyclotron Resonance (FT-ICR) method. New developments for both techniques are described, which will improve the detection efficiency and the suppression of contaminations in the case of TOF-ICR. The FT-ICR detection systems will allow for the investigation of an incoming ion bunch from a radioactive-beam facility on the one hand, and for the detection of a single singly charged ion in the Penning trap on the other hand.     

Synthesis and Antitumor,Antioxidant Effects Studies of N-Ethylpiperazine Substitute Thiourea Ligands and Their Copper(II) Complexes

A series of N-ethylpiperazine substitute thioureas [C₆N₂H₁₃NHCSNHR], where R = -C₃H₅ (L ₁ ), -C₁₀H₇ (L ₂ ), and -C₇H₇ (L ₃ ), and their copper (II) complexes have been synthesized. These ligands and complexes have been characterized by elemental analyses, IR, ¹H and ¹³C-NMR spectra, UV-Vis, magnetic susceptibility, thermogravimetrical analyses, and MALDI-TOF MS. In vitro antitumor activity of ligands and their complexes has been screened toward several tumor cell lines. The effects on these complexes of the growth of L1210 and MCF7 were studied comparatively with that of free ligands. Antioxidant and radical scavenging activities of synthesized compounds were determined by various in vitro assays including 1,1-diphenyl-2-picryl-hydrazyl free radicals (DPPH), 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid radicals (ABTS+), and ferrous ion (Fe²⁺) chelating activities. Moreover, these activities were compared to synthetic and standard antioxidant trolox. The results showed that the synthesized compounds had effective antioxidant power.     

Simultaneous Determination of Plasma Enrichments of 1-13C- and 15N-Labelled Phenylalanine and Tyrosine

Abstract

A methylchloroformate derivative was used for the simultaneous determination of plasma enrichments of 1-¹³C-phenylalanine, 1-¹³C-tyrosine, ¹⁵N-phenylalanine and ¹⁵N-tyrosine by gas chromatography/mass spectrometry. All four tracer enrichments could be measured in a single GC run. A specific ion fragment was obtained for each tracer. This approach allowed an easy determination of the “tracer to tracee ratios”. Each ion fragment could be measured with an appropriate single-to-noise ratio and precision in samples obtained from 100 μl plasma. The derivatization consists of a fast one-step reaction. Therefore it is well suited for studies involving a large number of samples, such as non-steady state bolus studies.     

Photo-induced reactions of hemin (DMSO) n clusters (n = 0-3) produced with electrospray ionization

Photo-induced reaction of [Fe(III)-protoporphyrin]⁺ (hemin⁺) ions solvated with dimethylsulfoxide (DMSO) is investigated by using a tandem mass spectrometer with electrospray ionization. We measure the photodissociation yields of mass-selected hemin⁺(DMSO)_n clusters for n = 0-3 in the energy region of 15 800-28 200 cm^-1. The mass spectra of the fragment ions show the β-cleavage of carboxymethyl groups in addition to the evaporation of solvent molecules. Yield of the β-cleavage reaction is found to depend strongly on the excitation energy and the number of solvent molecules. We also examine the metastable decomposition of the clusters following primary mass selection and determine the incremental solvent binding energies and internal energies for the clusters using evaporative ensemble model. From these results, we investigate the reaction mechanism of β-cleavage of hemin⁺ ion.     

A Multifunctional Fluorescence Probe for the Detection of Cations in Aqueous Solution: the Versatility of Probes Based on Peptides

We synthesized a tetra-functional fluorescence probe based on dansyl and peptide motif, dansyl-Gly-Trp (DGT, 1), that efficiently bound several metal ions and showed distinguishing optical properties. The probe 1 could respond to Hg²⁺ with enhanced and blue-shifted fluorescence emission but to Cu²⁺ with obvious fluorescence quenching. In addition, 1 was sensitive to pH ranging from 2.0 to 5.0 and precipitated in the presence of Pb²⁺ at neutral conditions. The combination of these intrinsic properties with the selective responses to different chemical inputs allows this system to be implemented as an ionic switch. Furthermore, 1 could penetrate the cell membrane and accumulated well in intracellular region. The underlying mechanisms of the probe to different kind of metal ion were explored successfully by using either ¹H NMR, NOESY, electron paramagnetic resonance (EPR) or FT-IR spectra. In addition, to investigate the binding model of 1/Hg²⁺ and 1/Cu²⁺, simulations were also performed by using density functional theory (DFT) and reasonable binding configurations were achieved for these two complexes.     

Simultaneous SIMS and EID investigation on the interaction of oxygen with a W (100) surface

A UHV system, containing a beatable tungsten ribbon target (showing [100] planes), an ion source (Ar⁺, 2 keV) with mass separator, an electron source (300 eV), a quadrupole secondary ion mass filter, and a quadrupole gas analyzer is used for the study of the interaction of O₂ with W (100) by simultaneous, i.e. fast interchanging, “static” SIMS (secondary [ion-induced] ion mass spectrometry) and EID (electron-induced [ion] desorption). Two different adsorptive binding states can be distinguished: β₂ and β₁. The O⁺ emission cross section under electron bombardment from the β₂ state is smaller by a factor of about 10³ than from β₁ and is found to be temperature-dependent. After the state β₂ has been saturated and before the occupation of β₁ begins, an oxide formation process starts. This oxidation can be interpreted by a two-stage model.     

Atomic spectrometry and atomic mass spectrometry in bioanalytical chemistry

Abstract

Atomic spectrometry and atomic mass spectrometric (MS) techniques have been playing crucial roles in the field of biosciences. They detect elements with relatively high sensitivities and are thus applicable to a wide range of analytical targets. In the past decade, determination of bio-relevant metallic elements continues to be of interest, while particularly noteworthy are methods developed for small molecules, peptides, proteins, nucleic acids and even cells that well exploited the bio-analytical strengths of atomic spectrometry and atomic MS, either in a direct or indirect manner. Quantitation, as well as speciation and imaging analyses are all involved. The present review aims to assimilate recent advances in bio-analysis utilizing atomic spectrometry and atomic MS, primarily covering the period of 2013–2018, in an attempt to provide readers insight into the developing trends of this research frontier. Followed by concluding remarks and perspectives, the applications are divided into the following four catalogs: (i) toxicologically important metal-containing species, with an emphasis on quantitative and imaging analysis; (ii) quantitation of biomolecules using naturally occurring heteroatoms; (iii) exogenous metal ion or nanoparticle tagging-based strategies in bioassays; and (iv) label-free detection of biomolecules.     

Comparison of the Stability of Calix[4]arene‐crown‐6‐cation Binary Complexes Under Electrospray Mass Spectrometry

Electrospray/mass spectrometry ESI/MS analyses were performed to study the stability of calix[4]‐arene‐crown‐6/alkali cation complexes in the gas phase, and in acetonitrile/water mixtures. This approach allowed a comparison with previous investigations by molecular‐dynamic simulations,which demonstrated a complementarity between calculation and experiment. Experimental results obtained from ESI/MS confirm that the stability of calixarene/cation complexes depends upon the medium used. Indeed, the calixarene in solution presents a strong affinity for cesium, whereas in the gas phase, it has a stronger affinity for sodium. Similarly, the stability of [calixarene + Na]⁺‐type complexes in the solvent phase is increased by the presence of water in the dilution system (up to 40% in acetonitrile), whereas other alkaline complexes are destabilized by water in any proportion. Finally, calixarenes that bear benzo groups on their crowns have an affinity for sodium, which is weak in solution, but considerably stronger in the gas phase.