Massenspektrometrie - Neueste Forschungsartikel der Fachverlage
Die Urheberrechte und
Veroeffentlichungsrechte der in der nachfolgenden Liste aufgefuehrten
Fachartikel liegen bei den jeweiligen Verlagen, die am Ende des
jeweiligen Artikels
als Quelle genannt werden. Diese sind auch fuer die Inhalte
verantwortlich.
Hinweise zur Veroeffentlichung Ihrer
Pressmitteilung unter Internetchemie.Info entnehmen Sie bitte der
entsprechenden Info-Seite .
Diese Seite koennen Sie mit folgender Tastenkombination nach Stichwoertern
durchsuchen: <STRG > und <F >.
Auf dieser Seite
beruecksichtige naturwissenschaftliche Journale:
Mass Spectrometry Reviews - published by
Wiley Interscience -
Journal of Mass Spectrometry - published by
Wiley Interscience -
Aktuelle wissenschaftliche Fachartikel der
genannten Journale:
Analysis of carbohydrates and glycoconjugates by matrix-assisted laser desorption/ionization mass spectrometry: An update for 2003-2004
This review is the third update of the original review, published in 1999, on the application of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings the topic to the end of 2004. Both fundamental studies and applications are covered. The main topics include methodological developments, matrices, fragmentation of carbohydrates and applications to large polymeric carbohydrates from plants, glycans from glycoproteins and those from various glycolipids. Other topics include the use of MALDI MS to study enzymes related to carbohydrate biosynthesis and degradation, its use in industrial processes, particularly biopharmaceuticals and its use to monitor products of chemical synthesis where glycodendrimers and carbohydrate-protein complexes are highlighted. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Characterization of natural wax esters by MALDI-TOF mass spectrometry
The applicability of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) to the analysis of wax esters (WEs) was investigated. A series of metal salts of 2,5-dihydroxybenzoic acid (DHB) was synthesized and tested as possible matrices. Alkali metal (Li, Na, K, Rb, Cs) and transition metal (Cu, Ag) salts were studied. The matrix properties were evaluated, including solubility in organic solvents, threshold laser power that should be applied for successful desorption/ionization of WEs, the nature of the matrix ions and the mass range occupied by them, and the complexity of the isotope clusters for individual metals. Lithium salt of dihydroxybenzoic acid (LiDHB) performed the best and matrices with purified lithium isotopes (6LiDHB or 7LiDHB) were recommended for WEs. Three sample preparation procedures were compared: (1) mixing the sample and matrix in a glass vial and deposition of the mixture on a MALDI plate (Mix), (2) deposition of sample followed by deposition of matrix (Sa/Ma), and (3) deposition of matrix followed by deposition of sample (Ma/Sa). Morphology of the samples was studied by scanning electron microscopy. The best sample preparation technique was Ma/Sa with the optimum sample to matrix molar ratio 1 : 100. Detection limit was in the low picomolar range. The relative response of WEs decreased with their molecular weight, and minor differences between signals of saturated and monounsaturated WEs were observed. MALDI spectra of WEs showed molecular adducts with lithium [M + Li]+. Fragments observed in postsource decay (PSD) spectra were related to the acidic part of WEs [RCOOH + Li]+ and they were used for structure assignment. MALDI with LiDHB was used for several samples of natural origin, including insect and plant WEs. A good agreement with GC/MS data was achieved. Moreover, MALDI allowed higher WEs to be analyzed, up to 64 carbon atoms in Ginkgo biloba leaves extract. Copyright © 2008 John Wiley & Sons, Ltd.
The role of mass spectrometry in atomic weight determinations
The 1914 Nobel Prize for Chemistry was awarded to Theodore Richards, whose work provided an insight into the history of the birth and evolution of matter as embedded in the atomic weights. However, the secret to unlocking the hieroglyphics contained in the atomic weights is revealed by a study of the relative abundances of the isotopes. A consistent set of internationally accepted atomic weights has been a goal of the scientific community for over a century. Atomic weights were originally determined by chemical stoichiometry - the so-called "Harvard Method," but this methodology has now been superseded by the "physical method," in which the isotopic composition and atomic masses of the isotopes comprising an element are used to calculate the atomic weight with far greater accuracy than before. The role of mass spectrometry in atomic weight determinations was initiated by the discovery of isotopes by Thomson, and established by the pioneering work of Aston, Dempster, and Nier using sophisticated mass spectrographs. The advent of the sector field mass spectrometer in 1947, revolutionized the application of mass spectrometry for both solids and gases to other fields of science including atomic weights. Subsequently, technological advances in mass spectrometry have enabled atomic masses to be determined with an accuracy better than one part in 107, whilst the absolute isotopic composition of many elements has been determined to produce accurate values of their atomic weights. Conversely, those same technological developments have revealed significant variations in the isotope abundances of many elements caused by a variety of physiochemical mechanisms in natural materials. Although these variations were initially seen as an impediment to the accuracy with which atomic weights could be determined, it was quickly realized that nature had provided a new tool to investigate physiochemical and biogeochemical mechanisms in nature, which could be exploited by precise and accurate isotopic measurements. Atomic weights can no longer be regarded as constants of nature, except for the monoisotopic elements whose atomic weights are determined solely by the relative atomic mass of that nuclide. Stable isotope geochemists developed mass spectrometric protocols by the adoption of internationally accepted reference materials for the light elements, to which measurements from various laboratories could be compared. Subsequently, a number of heavy elements such as iron, molybdenum and cadmium have been shown to exhibit isotope fractionation. The magnitude of such isotope fractionation in nature is less than for the light elements, but technological developments, such as multiple collector-inductively coupled plasma-mass spectrometry, have enabled such fractionation effects to be determined. Measurements of the atomic weights of certain elements affect the determination of important fundamental constants such as the Avogadro Constant, the Faraday Constant and the Universal Gas Constant. Heroic efforts have been made to refine the accuracy of the atomic weight of silicon, with the objective of replacing the SI standard of mass - the kilogram - with the Avogadro Constant. Improvements in these fundamental constants in turn affect the set of self-consistent values of other basic constants through a least-squares adjustment methodology. Absolute isotope abundances also enable the Solar System abundances of the s-, r-, and p-process of nucleosynthesis to be accurately determined, thus placing constraints on theories of heavy element nucleosynthesis. Future developments in the science of atomic weight determinations are also examined. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Metal-ligand solution equilibria studied by electrospray ionization mass spectrometry: effect of instrumental parameters
Electrospray ionization mass spectrometry (ESI-MS) is being increasingly employed in the study of metal-ligand equilibria in aqueous solution. In the present work, the ESI-MS spectral changes due to different settings of the following instrumental parameters are analyzed: the solution flow rate (FS), the nebulizer gas flow rate (FG), the sprayer potential (E), and the temperature of the entrance capillary (T). Twenty-eight spectra were obtained for each of six samples containing aluminum(III) and 2,3-dihydroxypyridine at various pH, in the absence or in the presence of a buffer and of sodium ions. Among the considered instrumental parameters, T produced the largest effects on the ionic intensities. FS and FG affected the ESI-MS spectra to a lower extent than T. In the investigated conditions E had the weakest effects on the spectra.The correlations observed between the ionic intensities and these instrumental parameters were interpreted considering the presence of three kinds of perturbations occurring in the ESI-MS ion source: formation of some dimers in the droplets, different transfer efficiencies from the droplets to the gas phase for different complexes (according to their surface activity), and subsequent partial thermal decomposition of the dimers and of one of the monomeric complexes in the gas phase. Our results show that the evaluation of the effects produced in the ESI-MS spectra by a change of instrumental parameters can allow to identify the perturbations occurring when metal-ligand solutions are studied by ESI-MS. Copyright © 2008 John Wiley & Sons, Ltd.
Application of negative ion MS/MS to the identification of N-glycans released from carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1)
Structures of N-glycans released from rat CEACAM1 expressed in human embryonic kidney cells were determined by MALDI and negative ion nanospray MS/MS techniques. The major carbohydrates were bi-, tri- and tetra-antennary complex glycans with and without sialic acid, fucose and bisecting GlcNAc residues. High-mannose glycans, predominantly Man5GlcNAc2, were also found. The negative ion fragmentation technique easily identified the branching pattern of the triantennary glycans (mainly branched on the 6-antenna) and the presence of 'bisecting' GlcNAc residues (attached to the 4-position of the core mannose), features that are difficult to determine by traditional techniques. Sialic acids were in both [alpha]2-3 and [alpha]2-6 linkage as determined by MALDI-TOF MS following linkage-specific derivatization. Copyright © 2008 John Wiley & Sons, Ltd.
Mass spectrometry analysis of the influenza virus
The role of mass spectrometry to probe characteristics of the influenza virus, and vaccine and antiviral drugs that target the virus, are reviewed. Genetic and proteomic approaches have been applied which incorporate high resolution mass spectrometry and mass mapping to genotype the virus and establish its evolution in terms of the primary structure of the surface protein antigens. A mass spectrometric immunoassay has been developed and applied to assess the structure and antigenicity of the virus in terms of the hemagglutinin antigen. The quantitation of the hemagglutinin antigen in vaccine preparations has also been conducted that is of importance to their efficacy. Finally, the characterization and quantitation of antiviral drugs against the virus, and their metabolites, have been monitored in blood, serum, and urine. The combined approaches demonstrate the strengths of modern mass spectrometric methods for the characterization of this killer virus. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Analysis of macrolide antibiotics, using liquid chromatography-mass spectrometry, in food, biological and environmental matrices
Macrolides are a group of antibiotics that have been widely used in human medical and veterinary practices. Analysis of macrolides and related compounds in food, biological, and environmental matrices continue to be the focus of scientists for the reasons of food safety, pharmacokinetic studies, and environmental concerns. This article presents an overview on the primary biological properties of macrolides and their associated analytical issues, including extraction, liquid chromatography-mass spectrometry (LC-MS), method validation, and measurement uncertainty. The main techniques that have been used to extract macrolides from various matrices are solid-phase extraction and liquid-liquid extraction. Conventional liquid chromatography (LC) with C18 columns plays a dominant role for the determination of macrolides, whereas ultra-performance liquid chromatography (UPLC) along with sub-2 µm particle C18 columns reduces run time and improves sensitivity. Mass spectrometry (MS), serving as a universal detection technique, has replaced ultraviolet (UV), fluorometric, and electrochemical detection for multi-macrolide analysis. The triple-quadrupole (QqQ), quadrupole ion trap (QIT), triple-quadrupole linear ion trap, time-of-flight (TOF), and quadrupole time-of-flight (QqTOF) mass spectrometers are current choices for the determination of macrolides, including quantification, confirmation, identification of their degradation products or metabolites, and structural elucidation. LC or UPLC coupled to a triple-quadrupole mass spectrometer operated in the multiple-reaction monitoring (MRM) mode (LC/MS/MS) is the first choice for quantification. UPLC-TOF or UPLC-QqTOF has been recognized as an emerging technique for accurate mass measurement and unequivocal identification of macrolides and their related compounds. © 2008 Crown in the right of Canada. Published by John Wiley & Sons, Inc., Mass Spec Rev
Allergenic compounds on the inner and outer surfaces of natural latex gloves: MALDI mass spectrometry and imaging of proteinous allergens
Natural latex gloves are the cause of a severe health problem to an increasing number of healthcare workers or patients due to the presence of protein allergens as Hevein or Rubber Elongation Factor (REF). One of the most challenging problems is the in situ localization of theses allergens in, e.g. gloves, to estimate the allergenic potential of the latex material. A sample preparation protocol applying a binary matrix-assisted laser desorption/ionization(MALDI) matrix containing [alpha]-cyano-4-hydroxy cinnamic acid (CHCA) and 2,5-dihydroxy benzoic acid (DHB) on trifluoro acetic acid (TFA) etched latex glove surfaces allowed the direct determination (exact molecular weight) of Hevein, REF and a truncated form of REF (tREF) within nine different brands of natural latex gloves by means of MALDI-TOF-MS in the linear mode. MALDI mass spectrometry demonstrated that Hevein, tREF and REF were present on the inner surfaces (in direct contact with the skin) of many, but not all, investigated gloves without any prior extraction procedure. Additionally, different isoforms of the allergen Hevein were detected (exhibiting ragged C-termini). tREF and REF could always be detected beside each other, but were not observed on every latex glove sample, which contained Hevein. It was also demonstrated that there is a significant difference in terms of proteins and polymers between inner and outer surfaces of gloves, which helps to explain the different allergenic potential of these.MALDI imaging allowed for the first time the unambiguous localization of all three allergens in parallel and showed that Hevein was present on 36% of the investigated area of a latex glove with a certain localization, whereupon, tREF and REF were only found on 25% of the investigated material. Copyright © 2008 John Wiley & Sons, Ltd.
Mass spectrometry in biodefense
Potential agents for biological attacks include both microorganisms and toxins. In mass spectrometry (MS), rapid identification of potential bioagents is achieved by detecting the masses of unique biomarkers, correlated to each agent. Currently, proteins are the most reliable biomarkers for detection and characterization of both microorganisms and toxins, and MS-based proteomics is particularly well suited for biodefense applications. Confident identification of an organism can be achieved by top-down proteomics following identification of individual protein biomarkers from their tandem mass spectra. In bottom-up proteomics, rapid digestion of intact protein biomarkers is again followed by MS/MS to provide unambiguous bioagent identification and characterization. Bioinformatics obviates the need for culturing and rigorous control of experimental variables to create and use MS fingerprint libraries for various classes of bioweapons. For specific applications, MS methods, instruments and algorithms have also been developed for identification based on biomarkers other than proteins and peptides. Copyright © 2008 John Wiley & Sons, Ltd.
Screening and characterization of reactive metabolites using glutathione ethyl ester in combination with Q-trap mass spectrometry
The present study describes a new analytical approach for the detection and characterization of chemically reactive metabolites using glutathione ethyl ester (GSH-EE) as the trapping agent in combination with hybrid triple quadrupole linear ion trap mass spectrometry. Polarity switching was applied between a negative precursor ion (PI) survey scan and the positive enhanced product ion (EPI) scan. The negative PI scan step was carried out monitoring the anion at m/z 300, corresponding to deprotonated [gamma]-glutamyl-dehydroalanyl-glycine ethyl ester originating from the GSH-EE moiety. Samples resulting from incubations in the presence of GSH-EE were cleaned and concentrated by solid-phase extraction, followed by the PI-EPI analysis. Unambiguous identification of GSH-EE-trapped reactive metabolites was greatly facilitated by the unique survey scan of the anion at m/z 300, which achieved less background interference, in particular, from endogenous glutathione adducts present in human liver microsomes. Further structural characterization was achieved by analyzing positive MS2 spectra that featured rich fragments without mass cutoff and were acquired in the same liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The effectiveness and reliability of this approach was evaluated using a number of model compounds in human liver microsomal incubations, including acetaminophen, amodiaquine, carbamazepine, 4-ethylphenol, imipramine and ticlopidine. In addition, iminoquinone reactive metabolites of mianserin were trapped and characterized for the first time using this method. Compared to neutral loss (NL) scanning assays using GSH as the trapping agent, the results have demonstrated superior selectivity, sensitivity, and reliability of this current approach. Copyright © 2008 John Wiley & Sons, Ltd.
Mass spectrometry for structural characterization of therapeutic antibodies
Antibodies, also known as immunoglobulins, have emerged as one of the most promising classes of therapeutics in the biopharmaceutical industry. The need for complete characterization of the quality attributes of these molecules requires sophisticated techniques. Mass spectrometry (MS) has become an essential analytical tool for the structural characterization of therapeutic antibodies, due to its superior resolution over other analytical techniques. It has been widely used in virtually all phases of antibody development. Structural features determined by MS include amino acid sequence, disulfide linkages, carbohydrate structure and profile, and many different post-translational, in-process, and in-storage modifications. In this review, we will discuss various MS-based techniques for the structural characterization of monoclonal antibodies. These techniques are categorized as mass determination of intact antibodies, and as middle-up, bottom-up, top-down, and middle-down structural characterizations. Each of these techniques has its advantages and disadvantages in terms of structural resolution, sequence coverage, sample consumption, and effort required for analyses. The role of MS in glycan structural characterization and profiling will also be discussed. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Oscore: a combined score to reduce false negative rates for peptide identification in tandem mass spectrometry analysis
Tandem mass spectrometry (MS/MS) has been widely used in proteomics studies. Multiple algorithms have been developed for assessing matches between MS/MS spectra and peptide sequences in databases. However, it is still a challenge to reduce false negative rates without compromising the high confidence of peptide identification. In this study, we developed the score, Oscore, by logistic regression using SEQUEST and AMASS variables to identify fully tryptic peptides. Since these variables showed complicated association with each other, combining them together rather than applying them to a threshold model improved the classification of correct and incorrect peptide identifications. Oscore achieved both a lower false negative rate and a lower false positive rate than PeptideProphet on datasets from 18 known protein mixtures and several proteome-scale samples of different complexity, database size and separation methods. By a three-way comparison among Oscore, PeptideProphet and another logistic regression model which made use of PeptideProphet's variables, the main contributor for the improvement made by Oscore is discussed. Copyright © 2008 John Wiley & Sons, Ltd.
Electrophilic aromatic substitution and single-electron transfer (SET) by the phenylium ion in the gas phase: characterization of a long-lived SET intermediate
Gas-phase mass spectrometric studies and calculations were performed for the reaction of naked phenylium ion with several benzene halides. From these reactions, the molecular ion for biphenyl as the predominant product was obtained only from the reaction of phenylium ions with iodobenzene and bromobenzene. Furthermore, through the collision-induced dissociation (CID) of the ion at m/z 281, the only dissociation observed is the loss of a phenyl radical, which indicates that a single-electron transfer (SET) mechanism might have occurred within the reaction. Additionally, according to the comparison between the CID experiments of those isomeric compounds of the [sigma]-complexes and the CID experiment of the ion at m/z 281 captured in the ion trap, we have also defined the captured ion at m/z 281 as an SET-intimate ion pair rather than those of [sigma]-complexes or the diphenyliodonium. Copyright © 2008 John Wiley & Sons, Ltd.
A kinetic study of guest displacement reactions on a host-guest complex with a photoswitchable calixarene
The displacement processes of several guests, incorporated in a calixarene host system, were investigated in the gas phase by electrospray ionization-Fourier transform-ion cyclotron resonance (ESI-FT-ICR) mass spectrometry. The complexes resulting from a resorcin[4]arene host with ammonia and sec-butylamine guests were isolated in an ICR-cell, separately using both states of the photoswitch as well as two reference systems for the open and closed forms of the photoswitchable host. The isolated complexes were forced to exchange the guest by using methylamine, ethylamine and sec-butylamine, resulting in different reaction rates for all the measured systems. Especially, the reaction rates of both states of the photoswitch are dependent on the provided guest. Potential side effects like proton exchanges were examined by an H/D-exchange experiment. The results were investigated and supported by quantum chemical calculations (DFT). Copyright © 2008 John Wiley & Sons, Ltd.
A fast screening MALDI method for the detection of cocaine and its metabolites in hair
Matrix-assisted laser desorption/ionisation (MALDI) mass spectrometry was used for the rapid detection of cocaine, benzoylecgonine and cocaethylene in hair. Different MALDI sample preparation procedures have been tested and the employment of a multi-layer 'graphite-sample-electrosprayed [alpha]-cyano-4-hydroxycinnamic acid (HCCA)' yielded the best results for standard solutions of the target analytes. The same approach was subsequently applied to hair samples that were known to contain cocaine, benzoylecgonine and cocaethylene, as determined by a classical GC-MS method. It was however necessary to extract hair samples by incubating them in methanol/trifluoroacetic acid for a short time (15 min) at 45 °C; 1 µl of the obtained supernatant was deposed on a metal surface treated with graphite, and HCCA was electrosprayed on it. This procedure successfully suppressed matrix peaks and was effective in detecting all the target analytes as their protonated species. The results obtained give further confirmation of the effectiveness of the MALDI for detecting drugs and their metabolites in complex biological matrices. The method can be useful as a fast screening procedure to detect the presence of cocaine and metabolites in hair samples. Copyright © 2008 John Wiley & Sons, Ltd.
Histidine-rich peptide: evidence for a single zinc-binding site on H5WYG peptide that promotes membrane fusion at neutral pH
The histidine-rich peptide H5WYG (GLFHAIAHFIHGGWHGLIHGWYG) was found to induce membrane fusion at physiologic pH in the presence of zinc chloride. In this study, we examined the ion selectivity of the interaction of Zn2+ with H5WYG. This investigation was conducted by using adsorption at air/water interface and mass spectrometry. We found that a peptide-metal complex is formed with Zn2+ ions. Electrospray ionisation-mass spectrometry (ESI-MS) reveals that the [H5WYG + Zn + 2H]4+, [H5WYG + Zn + H]3+ and [H5WYG + Zn]2+ ions, appearing by increasing the amount of Zn2+ equivalent, correspond to a monomolecular H5WYG - Zn2+ complex. Tandem mass spectrometry (MS/MS) provides evidence for the binding of the single Zn2+ ion to the H11 and H19 and probably H15 residues. Copyright © 2008 John Wiley & Sons, Ltd.
Characterization of saccharides and phenolic acids in the Chinese herb Tanshen by ESI-FT-ICR-MS and HPLC
This study sought to determine the main components (saccharides and phenolic acids) in crude extract of the Chinese herb Tanshen by electrospray ionization Fourier transform ion cyclotron resonant mass spectrometry (ESI-FT-ICR-MS) in negative-ion mode. Eleven compounds were identified as phenolic acids by exact mass measurement and further confirmed by sustained off-resonance irradiation (SORI) CID data. In addition, monosaccharides and oligosaccharides (n = 2 [sim] 5) and a serial of corresponding anionic adducts of saccharide were observed without adding any anions additionally to the extract solution, and the anionic components were unambiguously identified as H2O, HCl, HCOOH, HNO3, C3H6O2, H2SO4 and C5H7NO3 according to the exact mass measurement results. Furthermore, the saccharide types in Tanshen extract were identified as raffitrinose, saccharose, glucose, galactose and fructose with HPLC by comparing standards. Copyright © 2008 John Wiley & Sons, Ltd.
Theoretical and experimental studies of cationized uracil complexes in the gas phase
Cationized uracil clusters were generated in the gas phase by electrospray ionization (ESI). Mass spectrometry experiments showed that with particular experimental conditions, decameric uracil clusters are magic number clusters. MS/MS experiments demonstrated that the structure of these decameric uracil clusters depends substantially on the size and the charge of the cation. On the basis of the ab initio and density functional theory (DFT) quantum chemistry calculations, structures for these decameric clusters were proposed. These structures are in agreement with the experimental mass spectra of modified nucleobases. Theoretical calculations showed that complexes experimentally observed using ESI-MS techniques, are not naturally the most stable in the gas phase. Copyright © 2008 John Wiley & Sons, Ltd.
Kinetic-energy-sensitive mass spectrometry for separation of different ions with the same m/z value
A double-focusing mass spectrometer (MS) equipped with a superconducting-tunnel-junction (STJ) detector has been applied to measure relative ionization cross-sections for the production of ions that are accompanied by different ion species with the same mass-to-charge (m/z) value. The STJ detector fabricated for this study enables kinetic energy (E) measurement of incoming individual ions at a counting rate of up to [sim]100 k ions/s and an energy resolution ([Delta]E/E) of 15%. Both high counting rate and high-energy resolution are necessary to independently determine both m and z and not the m/z value only in ion-counting MS experiments. Ions such as 14N22+ and 14N+ with the same m/z value can be clearly discriminated using a kinetic-energy-sensitive MS. This fine discrimination capability allows direct determination of relative ionization cross-sections of the homonuclear diatomic ions 14N22+/14N2+ and 16O22+/16O2+, which are difficult to measure due to the strong interference by the signals of their dissociated atomic ions with noticeably large ionization cross-sections. The new instrument requires no low-abundance heteronuclear diatomic molecules of the forms 14N15N or 16O17O to carry out ionization studies and thus, is expected to be useful in fields such as atmospheric science, interstellar science, or plasma physics. Copyright © 2008 John Wiley & Sons, Ltd.
Orbitrap mass spectrometry: Instrumentation, ion motion and applications
Since its introduction, the orbitrap has proven to be a robust mass analyzer that can routinely deliver high resolving power and mass accuracy. Unlike conventional ion traps such as the Paul and Penning traps, the orbitrap uses only electrostatic fields to confine and to analyze injected ion populations. In addition, its relatively low cost, simple design and high space-charge capacity make it suitable for tackling complex scientific problems in which high performance is required. This review begins with a brief account of the set of inventions that led to the orbitrap, followed by a qualitative description of ion capture, ion motion in the trap and modes of detection. Various orbitrap instruments, including the commercially available linear ion trap-orbitrap hybrid mass spectrometers, are also discussed with emphasis on the different methods used to inject ions into the trap. Figures of merit such as resolving power, mass accuracy, dynamic range and sensitivity of each type of instrument are compared. In addition, experimental techniques that allow mass-selective manipulation of the motion of confined ions and their potential application in tandem mass spectrometry in the orbitrap are described. Finally, some specific applications are reviewed to illustrate the performance and versatility of the orbitrap mass spectrometers. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Review of factors that influence the abundance of ions produced in a tandem mass spectrometer and statistical methods for discovering these factors
Proteomic technologies are important because they link genes, proteins and disease. The identification of proteins and peptides has been revolutionized in the last decade by the use of mass spectrometry. This method is highly sensitive and much faster than the chemical reactions used previously because it can fragment peptides in seconds rather than in hours or days. Proteins are digested with an enzyme, usually trypsin, and the resulting peptides are fragmented in a tandem mass spectrometer (MS/MS). The masses of the fragment ions formed in the MS/MS can be used to identify the sequence of amino acids in the peptides. However, a number of different factors have been found to influence the amount of the various types of fragment ion formed. In this article, we review these factors and their interrelation together with the statistical methods used to discover them. Information on the number of fragment ions formed is at present underused in peptide identification algorithms, and fully utilizing this information could improve current algorithms. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Mass spectrometric characterization of naphthenic acids in environmental samples: A review
There is a growing need to develop mass spectrometric methods for the characterization of oil sands naphthenic acids (structural formulae described by CnH2n+zO2 where n is the number of carbon atoms and "z" is referred to as the "hydrogen deficiency" and is equal to zero, or is a negative, even integer) present in environmental samples. This interest stems from the need to better understand their contribution to the total acid number of oil sands acids; along with assessing their toxicity in aquatic environments. Negative-ion electrospray ionization has emerged as the analytical technique of choice. For infusion samples, matrix effects are particularly evident for quantification in the presence of salts and co-elutants. However, such effects can be minimized for methods that employ chromatographic separation prior to mass spectrometry (MS) detection. There have been several advances for accurate identification of classes of naphthenic acid components that employ a range of MS hyphenated techniques. General trends measured for degradation of the NAs in the environment appear to be similar to those obtained with either low- or high-resolution MS. Future MS research will likely focus on (i) development of more reliable quantitative methods that use chromatography and internal standards, (ii) the utility of representative model naphthenic acids as surrogates for the complex NA mixtures, and (iii) development of congener-specific analysis of the principal toxic components. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Negative-mode MALDI mass spectrometry for the analysis of pigments using tetrathiafulvalene as a matrix
Negative-mode matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) analysis of industrial pigments was performed using tetrathiafulvalene (TTF), an electron donating compound, as a matrix. Comparing with other matrices such as tetracyanoquinodimethane (TCNQ), dithranol, 2-hydroxy-6-nitroaniline (HNA), and harmine hydrochloride, the most abundant analyte ions were produced by the TTF as radical or deprotonated anions. In addition, fragment ions in the negative mode of operation are also found to be much less abundant than those in the positive mode of operation. Copyright © 2008 John Wiley & Sons, Ltd.
Femtosecond laser ablation inductively coupled plasma mass spectrometry: Fundamentals and capabilities for depth profiling analysis
Laser ablation coupled to inductively coupled plasma mass spectrometry has become a versatile and powerful analytical method for direct solid analysis. The applicability has been demonstrated on a wide variety of samples, where major, minor, and trace element concentrations or isotope ratio determinations have been of interest. The pros and cons of UV-nsec laser ablation have been studied in detail, and indicate that aerosol generation, aerosol transport, and aerosol excitation-ionization within the ICP contribute to fractionation effects, which prevent this method from a more universal application to all matrices and all elements. Recent progresses in IR-fs and UV-fs laser ablation coupled to ICP-MS have been reported, which increase the inter-matrix and multi-element quantification capabilities of this method. These fundamental improvements in LA-ICP-MS are of significant importance for entering new applications in material science and related research fields. In particular, because coatings (conducting and non-conducting) consist of single or multilayers of various elemental composition and of different thickness (nm-mm range), significant progress in the field of depth profiling with fs-laser ablation can be expected. Therefore, in-depth profile analysis of polymers, semiconductors, and metal sample investigations, using ultra-fast laser ablation for sampling and the currently achievable figures of merit, are discussed. In this review manuscript, the enhanced capabilities of fs-LA-ICP-MS for direct solid sampling are highlighted, and it is discussed about current methods used for quantitative analysis and depth profiling, the ablation process of UV-ns and UV-fs, the influence of the laser beam profile, aerosol structure and transport efficiency, as well as the influence of the ICP-MS (e.g., vaporization and ionization efficiency in the plasma, and type of mass analyzer). © 2008 Wiley Periodicals, Inc. Mass Spectrom. Rev.
Mass spectrometry in systems biology: An overview
As an emerging field, systems biology is currently the talk of the town, which challenges our philosophy in comprehending biology. Instead of the reduction approach advocated in molecular biology, systems biology aims at systems-level understanding of correlations among molecular components. Such comprehensive investigation requires massive information from the "omics" cascade demanding high-throughput screening techniques. Being one of the most versatile analytical methods, mass spectrometry has already been playing a significant role at this early stage of systems biology. In this review, we documented the advances in modern mass spectrometry technologies as well as nascent inventions. Recent applications of mass spectrometry-based techniques and methodologies in genomics, proteomics, transcriptomics and metabolomics will be further elaborated individually. Undoubtedly, more applications of mass spectrometry in systems biology can be expected in the near future. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Sieve-based device for MALDI sample preparation. I. Influence of sample deposition conditions in oligonucleotide analysis to achieve significant increases in both sensitivity and resolution
Spraying of oligonucleotide-matrix solutions through a stainless steel (ss) sieve (38 µm, 450 mesh) leads to the formation, on the matrix-assisted laser desorption/ionization (MALDI) sample holder, of uniformly distributed microcrystals, well separated from each other. When the resulting sample holder surface is irradiated by laser, abundant molecular species form, with a clear increase in both intensity and resolution with respect to values obtained by 'Dried Droplet', 'Double Layer', and 'Sandwich' deposition methods. In addition, unlike the usual situation, the sample is perfectly homogeneous, and identical spectra are obtained by irradiating different areas. On one hand, the data indicate that this method is highly effective for oligonucleotide MALDI analysis, and on the other, that it can be validly employed for fully automated MALDI procedures. Copyright © 2008 John Wiley & Sons, Ltd.
Photoionization studies on various quinones by an infrared laser desorption/tunable VUV photoionization TOF mass spectrometry
Photoionization and dissociative photoionization characters of six quinones, including 1,2-naphthoquinone (1,2-NQ), 1,4-naphthoquinone (1,4-NQ), 9,10-phenanthroquinone (PQ), 9,10-anthraquinone (AQ), benz[a]- anthracene-7,12-dione (BAD) and 1,2-acenaphthylenedione (AND) have been studied with an infrared laser desorption/tunable synchrotron vacuum ultraviolet (VUV) photoionization mass spectrometry (IR LD/VUV PIMS) technique. Mass spectra of these compounds are obtained at different VUV photon energies. Consecutive losses of two carbon monoxide (CO) groups are found to be the main fragmentation pathways for all the quinones. Detailed dissociation processes are discussed with the help of ab initio B3LYP calculations. Ionization energies (IEs) of these quinones and appearance energies (AEs) of major fragments are obtained by measuring the photoionization efficiency (PIE) spectra. The experimental results are in good agreement with the theoretical data. Copyright © 2008 John Wiley & Sons, Ltd.
Plant proteomics: Concepts, applications, and novel strategies for data interpretation
Proteomics is an essential source of information about biological systems because it generates knowledge about the concentrations, interactions, functions, and catalytic activities of proteins, which are the major structural and functional determinants of cells. In the last few years significant technology development has taken place both at the level of data analysis software and mass spectrometry hardware. Conceptual progress in proteomics has made possible the analysis of entire proteomes at previously unprecedented density and accuracy. New concepts have emerged that comprise quantitative analyses of full proteomes, database-independent protein identification strategies, targeted quantitative proteomics approaches with proteotypic peptides and the systematic analysis of an increasing number of posttranslational modifications at high temporal and spatial resolution. Although plant proteomics is making progress, there are still several analytical challenges that await experimental and conceptual solutions. With this review I will highlight the current status of plant proteomics and put it into the context of the aforementioned conceptual progress in the field, illustrate some of the plant-specific challenges and present my view on the great opportunities for plant systems biology offered by proteomics. © 2008 Wiley Periodicals, Inc. Mass Spectrom. Rev.
Mass spectrometry-based proteomics in reproductive medicine
The emergence of powerful mass spectrometry-based proteomic techniques has added a new dimension to the field of biomedical research. Application of these high throughput methodologies in pregnancy-related pathology has contributed to the comprehension of the underlying pathophysiologies and the successful identification of relevant protein biomarkers that can potentially change early diagnosis and treatment of several medical conditions related to human pregnancy. Most of the existing research on human reproduction and gestation has focused on follicular fluid, cervical/vaginal fluid, and amniotic fluid. Although proteome technologies in reproductive medicine research are not as yet widely applied, characterization of the proteome of reproductive fluids can be expected to significantly improve maternal healthcare. This article aims to summarize the applications of mass spectrometry based technology on the most important and specific biological fluids related to reproduction and gestation. © 2008 Wiley Periodicals, Inc. Mass Spectrom. Rev.
Live single-cell video-mass spectrometry for cellular and subcellular molecular detection and cell classification
The molecular content from the cytoplasm of a live, single mammalian cell and its organelle were trapped with a nano-electrospray ionization (ESI) tip acting as a micropipette under a video microscope, and hundreds of small molecular peaks were detected by direct nano-ESI mass spectrometry (MS). Granule- or cytoplasm-specific peaks in a mast cell (RBL 2H3) model were extracted by paired t-test to demonstrate their specific localization. Some of the typical and specific molecules were successfully identified by MS/MS analysis. This method was also applied to the cell classification of seven types of cell lines at the single-cellular level by principal component analysis (PCA), revealing seven clusters in the multivariate score plot. Copyright © 2008 John Wiley & Sons, Ltd.
Theoretical and experimental study of tropylium formation from substituted benzylpyridinium species
Fragmentation pathways of unsubstituted and substituted benzylpyridinium compounds were investigated using mass-analysed kinetic energy (MIKE) technique in combination with high level of quantum chemical calculations in the gas phase. Fast atom bombardment (FAB) source was used for ionisation of the studied compounds. The formation of both benzylium and tropylium species were investigated. Hybrid Hartree-Fock/Density Functional Theory calculations have been performed to assess the geometries and the energies of the transition states and intermediates. For each cases, different reaction pathways were investigated, and particularly in the case of the formation of tropylium species, the formation of the seven-membered ring before or after the loss of pyridine were studied. The effect of para-methyl and para-methoxy substituents on the activation energy of the rearrangement process to form thermodynamically stable tropylium compounds has been studied. Theoretical calculations showed competition between direct bond cleavage and rearrangement reactions to form benzylium and tropylium compounds, respectively. Experimental results also suggested that the rearrangement process takes place to yield stable tropylium under "soft ionisation techniques", such as FAB. Copyright © 2008 John Wiley & Sons, Ltd.
The McLafferty rearrangement in the Glu residue in a cyclic lipopeptide determined by Q-TOF MS/MS
The intraresidue rearrangement and loss of the side chain of the Glu residue was found through MS/MS analysis of both original and methanol-esterified lipopeptides. Both Glu and Asp residues in the cyclic lipopeptide were esterified. The MS/MS results showed that the loss of fragment 72 or 86 was induced by McLafferty-type rearrangement from the Glu or esterified Glu. The mechanism of loss of the Glu residue can be used to determine or to corroborate the existence of the Glu and to help understand the fragment formation in MS/MS. The cleavage mechanism and m/z intensities imply that the sodium ion was easier attached and the cleavage would easily occur at specific sites. Copyright © 2008 John Wiley & Sons, Ltd.
Targeted identification of phosphorylated peptides by off-line HPLC-MALDI-MS/MS using LC retention time prediction
Protein phosphorylation is a type of posttranslational modification which plays an important role in cell regulation and signal transduction. Because of its biological relevance, a considerable amount of interest has been paid to the development of efficient techniques for phosphopeptide analysis. Although advances in MS control have enabled the high-throughput discovery of proteins from limited amounts of sample, automated selection of MS/MS precursor ions based on intensity alone can significantly hamper the detection of low-abundance phosphopeptides. On the basis of the observation that the introduction of a phosphate moiety does not dramatically change peptide retention time in reverse-phase chromatography, phosphopeptide specific MS/MS fragmentation attempts based on LC retention time and m/z were evaluated using a standard protein mixture, then using in vitro phosphorylated myelin basic protein. Results indicated that the majority (98%) of phosphopeptides identified eluted within a ± 4-min window of the predicted LC elution time. While studies presented here are primarily proof of concept in nature, data suggest that the use of LC retention time prediction could be a valuable constraint for the identification of phosphopeptides within a set of off-line LC deposited sample spots. It is expected that the development of these methods will not only permit the targeted identification of protein phosphorylation sites but also allow the in-depth analysis of the dynamic events linked to the posttranslational modification. Copyright © 2008 John Wiley & Sons, Ltd.
Differences between collisionally activated and electron-transfer dissociations found for CH2X2(X = Cl, Br, and I) by using alkali-metal targets
High-energy collisionally activated dissociation (HE-CAD) and high-energy electron- transfer dissociation (HE-ETD) on collisions with alkali-metal targets (Cs, K, and Na) were investigated for CH2X2+ (X = Cl, Br, and I) ions by tandem mass spectrometry (MS/MS). In the HE-CAD spectra observed, peaks associated with CH2X+ ions formed by a loss of a halogen atom are always predominant regardless of precursor ions and target metals. The observation of the predominant CH2X+ ions is explained by the lowest energy levels of the fragments of CH2X+ + X among the possible fragment energy levels and internal-energy distribution in HE-CAD. In the charge-inversion spectra, relative peak intensities of the negative ions formed by HE-ETD strongly depend on the precursor ions and the target metals. While the CHCl2- ion was predominant in the spectra of CH2Cl2+ regardless of target species, the most intense peaks in those of CH2Br2+ and CH2I2+ were ascribed to either Br- or CH2Br- and either I- or I2-, respectively, depending on the target metals. The dependence of the relative intensities of the fragment ions by HE-ETD on the precursor ions and target species are discussed on the basis of the energy levels of the neutral fragments and the narrow internal-energy distribution resulting from the near-resonant neutralization. It was demonstrated that HE-ETD using the alkali-metal targets provided rich information on the dissociation of the neutral species. Copyright © 2008 John Wiley & Sons, Ltd.
IRMPD spectra of Gly·NH4+ and proton-bound betaine dimer: evidence for the smallest gas phase zwitterionic structures
Zwitterionic structures exist extensively in biological systems and the electric field resulting from zwitterion formation is the driving force for determination of the properties, function and activity of biological molecules, such as amino acids, peptides and proteins. It is of considerable interest and import to investigate the stabilization of zwitterionic structures in the gas phase. Infrared multiple photon dissociation (IRMPD) spectroscopy is a very powerful and sensitive technique, which may elucidate clearly the structures of both ions and ionic clusters in the gas phase, since it provides IR vibrational fingerprint information. The structures of the clusters of glycine and ammonium ion and of the betaine proton-bound homodimer have been investigated using IRMPD spectroscopy, in combination with electronic structure calculations. The experimental and calculated results indicate that zwitterionic structure of glycine may be effectively stabilized by an ammonium ion. This is the smallest zwitterionic structure of an amino acid to be demonstrated in the gas phase. On the basis of the experimental IRMPD and calculated results, it is very clear that a zwitterionic structure exists in the proton-bound betaine dimer. The proton is bound to one of the carboxylate oxygens of betaine, rather than being equally shared. Investigations of zwitterionic structures in the isolated state are essential for an understanding of the intrinsic characteristics of zwitterions and salt bridge interactions in biological systems. Copyright © 2008 John Wiley & Sons, Ltd.
Calibration laws based on multiple linear regression applied to matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry
Operation of any mass spectrometer requires implementation of mass calibration laws to translate experimentally measured physical quantities into a m/z range. While internal calibration in Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) offers several attractive features, including exposure of calibrant and analyte ions to identical experimental conditions (e.g. space charge), external calibration affords simpler pulse sequences and higher throughput. The automatic gain control method used in hybrid linear trap quadrupole (LTQ) FT-ICR-MS to consistently obtain the same ion population is not readily amenable to matrix-assisted laser desorption/ionization (MALDI) FT-ICR-MS, due to the heterogeneous nature and poor spot-to-spot reproducibility of MALDI. This can be compensated for by taking external calibration laws into account that consider magnetic and electric fields, as well as relative and total ion abundances. Herein, an evaluation of external mass calibration laws applied to MALDI-FT-ICR-MS is performed to achieve higher mass measurement accuracy (MMA). Copyright © 2008 John Wiley & Sons, Ltd.
Halohydrination of epoxy resins using sodium halides as cationizing agents in MALDI-MS and DIOS-MS
Halohydrination of epoxy resins using sodium halides as cationizing agents in matrix-assisted laser desorption/ionization (MALDI) and desorption ionization on porous silicon mass spectrometry (DIOS-MS) were investigated. Different mass spectra were observed when NaClO4 and NaI were used as the cationizing agents at the highest concentration of 10.0 mM, which is much higher than that normally used in MALDI-MS. MALDI mass spectra of epoxy resins using NaI revealed iodohydrination to occur as epoxy functions of the polymers. The halohydrination also occurred using NaBr, but not NaCl, due to the differences in their nucleophilicities. On the basis of the results of experiments using deuterated CD3OD as the solvent, the hydrogen atom source was probably ambient water or residual solvent, rather than being derived from matrices. Halohydrination also occurred with DIOS-MS in which no organic matrix was used; in addition, reduction of epoxy functions was observed with DIOS. NaI is a useful cationizing agent for changing the chemical form of epoxy resins due to iodohydrination and, thus, for identifying the presence of epoxy functions. Copyright © 2008 John Wiley & Sons, Ltd.
Recognizing [alpha]-, [beta]- or [gamma]-substitution in pyridines by mass spectrometry
A general mass spectrometric method able to recognize the site of substitution of monosubstituted pyridines is described. The method requires that the molecule under investigation forms, upon ionization and dissociation, the respective [alpha]-, [beta]- or [gamma]- pyridinium ion of m/z 78. Pyridinium ions are stable and common fragments of ionized and protonated pyridines and are found to function as appropriate structurally diagnostic fragment ions. They can be identified by their characteristic and nearly identical collision-induced dissociation behavior and distinguished by the combined use of two structurally diagnostic ion/molecule reactions with acetonitrile and 2-methyl-1,3-dioxolane. [alpha]-, [beta]- or [gamma]-substitution in pyridines can, therefore, be securely recognized via an MS-only method based on structurally diagnostic ions and by the inspection of a single molecule (no need for intracomparisons within the whole set of isomers). Copyright © 2008 John Wiley & Sons, Ltd.
HPTLC/DESI-MS imaging of tryptic protein digests separated in two dimensions
Desorption electrospray ionization mass spectrometry (DESI-MS) was demonstrated as a method to detect and identify peptides from two-dimensional separations of cytochrome c and myoglobin tryptic digests on ProteoChrom HPTLC Cellulose sheets. Data-dependent tandem mass spectra were acquired during lane scans across the TLC plates. Peptides and the corresponding proteins were identified using a protein database search software. Two-dimensional distributions of identified peptides were mapped for each separated protein digest. Sequence coverages for cytochrome c and myoglobin were 81 and 74%, respectively. These compared well with those determined using the more standard HPLC/ESI-MS/MS approach (89 and 84%, respectively). Preliminary results show that use of more sensitive instrumentation has the potential for improved detection of peptides with low Rf values and improvement in sequence coverage. However, less multiple charging and more sodiation were seen in HPTLC/DESI-MS spectra relative to HPLC/ESI-MS spectra, which can affect peptide identification by MS/MS. Methods to increase multiple charging and reduce the extent of sodiation are currently under investigation. Published in 2008 by John Wiley & Sons, Ltd.
Analysis of environmental stress response on the proteome level
Thousands of man-made chemicals are annually released into the environment by agriculture, transport, industries, and other human activities. In general, chemical analysis of environmental samples used to assess the pollution status of a specific ecosystem is complicated by the complexity of the mixture, and in some cases by the very low toxicity thresholds of chemicals present. In that sense, a proteomics approach, capable of detecting subtle changes in the level and structure of individual proteins within the whole proteome in response to the altered surroundings, has obvious applications in the field of ecotoxicology. In addition to identifying new protein biomarkers, it can also help to provide an insight into underlying mechanisms of toxicity. Despite being a comparatively new field with a number of caveats, proteomics applications have spread from microorganisms and plants to invertebrates and vertebrates, gradually becoming an established technology used in environmental research. This review article highlights recent advances in the field of environmental proteomics, mainly focusing on experimental approaches with a potential to understand toxic modes of action and to identify novel ecotoxicological biomarkers. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Cluster emission and chemical reactions in oxygen and nitrogen ices induced by fast heavy-ion impact
Two ices, O2 and a mixture of O2 and N2, are bombarded by 252Cf fission fragments (FF) ([sim]65 MeV at target surface); the emitted positive and negative secondary ions are analyzed by time-of-flight mass spectrometry (TOF-SIMS). These studies shall enlighten sputtering from planetary and interstellar ices. Three temperature regions in the 28-42-K range are analyzed: (1) before N2 sublimation, in which hybrid chemical species are formed, (2) before O2 sublimation, in which the TOF mass spectrum is dominated by low-mass (O2)p cluster ions and (3) after O2 sublimation, in which (N2)p or (O2)p cluster ions are practically inexistent. In the first region, four hybrid ion series are observed: NOn-1+, N2On-2±, and N4On-4-. In the second region, two positive and negative ion series are identified: (O2)pO± and (O2)pO2±. Their yield distributions are fitted by the sum of two decreasing exponentials, whose decay constants are the same for all series. It is observed that the cluster ion desorption from solid oxygen is very similar to that of other frozen gases, but its yield distribution oscillates with a three- or six-atom periodicity, suggesting O3 or 3O2 units in the cluster structure, respectively. Copyright © 2008 John Wiley & Sons, Ltd.
Investigation of double-stranded DNA/drug interaction by ESI/FT ICR: orientation of dissociations relates to stabilizing salt bridges
Noncovalent complexes of DNA and Hoechst 33258 were investigated by ESI-FT/ICR MS in various activation modes (collision-induced dissociation (CID), sustained off-resonance irradiation collision-induced dissociation (SORI-CID), infrared multiphoton dissociation (IRMPD) and electron detachment dissociation (EDD)). The binding selectivity of Hoechst 33258 was confirmed by the comparative study of its noncovalent association with different DNA sequences. The CID spectra of [ds + HO - 5H]5- obtained with a linear hexapole ion trap resulted in unzipping of the strands. This outcome is a clue to the drug-binding mode, shading light on the localization of the binding sites of Hoechst 33258 to the DNA sequence. The IRMPD and SORI-CID experiments mainly gave DNA backbone cleavages and internal fragment ions. From this result, information on the localization of the binding sites of Hoechst 33258 in the DNA sequence was obtained. No sodium cationization was observed on the DNA sequence ions although they were present on fragmentation of the duplex, indicating that the backbone cleavages were generated from the single strand associated with the Hoechst 33258 where the number of alkali cation is restricted. Under electron detachment (ED) conditions, multiple EDs were achieved for the [ds + HO - 5H]5- ion without any significant dissociation. The presence of drug appears to enhance the stability of the multiply charged system. It was proposed that the studied noncovalent complex involved the formation of zwitterions and consequently strong salt-bridge interactions between DNA and drug. Copyright © 2008 John Wiley & Sons, Ltd.
Low-energy collision-induced fragmentation of negative ions derived from diesters of aliphatic dicarboxylic acids made with hydroxybenzoic acids
Diesters of ortho-hydroxybenzoic acid (salicylic acid) made with glutaric, adipic, and pimelic acids are the monomers of some potential drug candidates for aspirin patches. Collision-induced dissociation (CID) spectra of negative ion derived from these compounds show a 120-Da 'neutral loss' specific to the ortho isomers. In contrast, the anions derived from diesters of meta- and para-hydroxybenzoic acids show a 138-Da loss for an elimination of elements of hydroxybenzoic acid by a charge-remote mechanism. Deuterium labeling studies confirmed that the hydrogen atom transferred for hydroxybenzoic acid loss originates specifically from the [alpha] position of the dicarboxylic acid moiety. Although all spectra showed a peak at m/z 137, a charge-mediated process specific for the ortho compounds renders it the most prominent peak in the spectra of ortho compounds. Appropriate deuterium labeling experiments demonstrated that the hydrogen atom transferred for the formation of the m/z 137 ion in ortho compounds is specifically derived from the [alpha] position of the dicarboxylic acid moiety. Copyright © 2008 John Wiley & Sons, Ltd.
Insights into virus capsid assembly from non-covalent mass spectrometry
The assembly of viral proteins into a range of macromolecular complexes of strictly defined architecture is one of Nature's wonders. Unraveling the details of these complex structures and the associated self-assembly pathways that lead to their efficient and precise construction will play an important role in the development of anti-viral therapeutics. It will also be important in bio-nanotechnology where there is a plethora of applications for such well-defined macromolecular complexes, including cell-specific drug delivery and as substrates for the formation of novel materials with unique electrical and magnetic properties. Mass spectrometry has the ability not only to measure masses accurately but also to provide vital details regarding the composition and stoichiometry of intact, non-covalently bound macromolecular complexes under near-physiological conditions. It is thus ideal for exploring the assembly and function of viruses. Over the past decade or so, significant advances have been made in this field, and these advances are summarized in this review, which covers the literature up to the end of 2007. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
Determination of apparent decomposition threshold energies of lithium adducts of acylglycerols using tandem mass spectrometry and a novel derived effective reaction path length approach
Apparent decomposition threshold energies for the fragmentation pathways of lithiated acylglycerols were experimentally determined by collisional activation in a quadrupole-hexapole-quadrupole (QhQ) mass spectrometer. A previously developed 'derived effective reaction path length' approach for predicting bond dissociation energies (BDEs) of simple dissociations of electrostatic complexes such as alkali metal adducts (Li+), or halide adducts (Cl-) of acylglycerols, was extended to predict covalent bond apparent decomposition threshold energies of lithium adducts of a mono-acylglycerol, a 1,2-diacylglycerol, and a 1,3-diacylglycerol. The ability of the model to treat relatively large ionic systems (e.g. more than 100 atoms) represents a huge advantage of this approach. The model's calculated apparent decomposition threshold energies (Ea) are used in conjunction with the method of energy-resolved mass spectrometry, employing breakdown graphs, to give a more complete quantitative description of the fragmentation processes. Calculated Ea values allowed ranking of the 1,2-diacylglycerol as more reactive than the 1,3-diacylglycerol; the mono-acylglycerol was ranked the least reactive. The method was applied to the low molecular weight product ions generally associated with the hydrocarbon series CnH2n+1+, where two separate pathways are deduced as contributing to the production of the abundant m/z 81 fragment ion. The favored ranking of the neutral losses of fatty acyl substituents for the 1,2-diacylglycerol was determined as: loss of lithium fatty acetate > loss of fatty acid > loss of fatty acyl chain as ketene. For the 1,3-diacylglycerol, the descending order of ease of neutral loss was: loss of fatty acyl ketene > loss of lithium fatty acetate > loss of fatty acid. The results of this study demonstrate that the newly developed method is general in nature, and it can be used for the measurement of covalent bond decomposition threshold energies, as well as for the previously documented electrostatic (noncovalent) bond energies. Copyright © 2008 John Wiley & Sons, Ltd.
The ProteoMiner and the FortyNiners: Searching for gold nuggets in the proteomic arena
The present review covers modern aspects of combinatorial peptide ligand libraries (CPLL), as used to analyze the "low-abundance proteome" in association with mass spectrometry. First, the capturing properties of baits of different lengths (from single amino acid to hexa-peptides) are described to show that a plateau is rapidly reached above a tetra-peptide in length, thus confirming the validity of having adopted hexapeptides for the considered application. The mechanism of interaction with proteins from very complex proteomes and the ability to decrease the dynamic concentration range is demonstrated with the help of mass spectrometry analysis. Examples are given on how treatment with CPLLs dramatically improves the detectability of peptides in mass spectrometry analysis, permitting detection of a very large number of proteins as compared with control, untreated samples. The use of complementary libraries is discussed with the aim to discover additional low-abundance species that escaped the first library. A discussion on the possibility to discover extremely rare gene products, and the quantitative aspect of the technology when associated with mass spectrometry is also provided. Some insights on the applications for hidden, low-abundance biomarkers are also presented. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
A tandem MS precursor-ion scan approach to identify variable covalent modification of albumin Cys34: a new tool for studying vascular carbonylation
We developed a liquid chromatography electrospray ionisation multi-stage mass spectrometry (LC-ESI-MS/MS) approach based on precursor-ion scanning and evaluated it to characterize the covalent modifications of Cys34 human serum albumin (HSA) caused by oxidative stress and reactive carbonyl species (RCS) adduction. HSA was isolated and digested enzymatically to generate a suitable-length peptide (LQQCPF) containing the modified tag residue. The resulting LQQCPF peptides were identified by LC-ESI-MS/MS in precursor-ion scan mode and further characterized in product-ion scan mode. The product ions for precursor-ion scanning were selected by studying the MS/MS fragmentation of a series of LQQCPF derivatives containing Cys34 modified with different [alpha],[beta]-unsaturated aldehydes and di and ketoaldehydes. We used a Boolean logic to enhance the specificity of the method: this reconstitutes a virtual current trace (vCT) showing the peaks in the three precursor-ion scans, marked by the same parent ion. The method was first evaluated to identify and characterize the Cys34 covalent adducts of HSA incubated with 4-hydroxy-hexenal, 4-hydroxy-trans-2-nonenal (HNE) and acrolein (ACR). Then we studied the Cys34 modification of human plasma incubated with mildly oxidized low-density lipoproteins (LDL), and the method easily identified the LQQCPF adducts with HNE and ACR. In other experiments, plasma was oxidized by 2,2'-azobis(2-amidinopropane) HCl (AAPH) or by Fe2+/H2O2. In both conditions, the sulfinic derivative of LQQCPF was identified and characterized, indicating that the method is suitable not only for studying RCS-modified albumin, but also to check the oxidative state of Cys34 as a marker of oxidative damage. Copyright © 2008 John Wiley & Sons, Ltd.
Formation of [b3 - 1 + cat]+ ions from metal-cationized tetrapeptides containing [beta]-alanine, [gamma]-aminobutyric acid or [epsiv]-aminocaproic acid residues
The presence and position of a single [beta]-alanine ([beta]A), [gamma]-aminobutyric acid ([gamma]ABu) or [epsiv]-aminocaproic acid (Cap) residue has been shown to have a significant influence on the formation of bn+ and yn+ product ions from a series of model, protonated peptides. In this study, we examined the effect of the same residues on the formation of analogous [b3 - 1 + cat]+ products from metal(Li+, Na+ and Ag+)-cationized peptides. The larger amino acids suppress formation of b3+ from protonated peptides with general sequence AAXG (where X = [beta]-alanine, [gamma]-aminobutyric acid or [epsiv]-aminocaproic acid), presumably because of the prohibitive effect of larger cyclic intermediates in the 'oxazolone' pathway. However, abundant [b3 - 1 + cat]+ products are generated from metal-cationized versions of AAXG. Using a group of deuterium-labeled and exchanged peptides, we found that formation of [b3 - 1 + cat]+ involves transfer of either amide or [alpha]-carbon position H atoms, and the tendency to transfer the atom from the [alpha]-carbon position increases with the size of the amino acid in position X. To account for the transfer of the H atom, a mechanism involving formation of a ketene product as [b3 - 1 + cat]+ is proposed. Copyright © 2008 John Wiley & Sons, Ltd.
Proteomics in gram-negative bacterial outer membrane vesicles
Gram-negative bacteria constitutively secrete outer membrane vesicles (OMVs) into the extracellular milieu. Recent research in this area has revealed that OMVs may act as intercellular communicasomes in polyspecies communities by enhancing bacterial survival and pathogenesis in hosts. However, the mechanisms of vesicle formation and the pathophysiological roles of OMVs have not been clearly defined. While it is obvious that mass spectrometry-based proteomics offers great opportunities for improving our knowledge of bacterial OMVs, limited proteomic data are available for OMVs. The present review aims to give an overview of the previous biochemical, biological, and proteomic studies in the emerging field of bacterial OMVs, and to give future directions for high-throughput and comparative proteomic studies of OMVs that originate from diverse Gram-negative bacteria under various environmental conditions. This article will hopefully stimulate further efforts to construct a comprehensive proteome database of bacterial OMVs that will help us not only to elucidate the biogenesis and functions of OMVs but also to develop diagnostic tools, vaccines, and antibiotics effective against pathogenic bacteria. © 2008 Wiley Periodicals, Inc., Mass Spec Rev
