Transform Ion Cyclotron Resonance MS Research Articles

Ferric particle-assisted LDI-MS platform for metabolic fingerprinting of diabetic retinopathy.

To explore the metabolic fingerprints of diabetic retinopathy (DR) in individuals with type 2 diabetes using a newly-developed laser desorption/ionization mass spectrometry (LDI-MS) platform assisted by ferric particles. Metabolic fingerprinting was performed using aferric particle-assisted LDI-MS platform. A nested population-based case-control study was performed on 216 DR cases and 216 control individuals with type 2 diabetes. DR cases and control individuals with type 2 diabetes were comparable for a list of clinical factors. The newly-developed LDI-MS platform allowed us to draw theblueprint of plasma metabolic fingerprints from participants with and without DR. The neural network afforded diagnostic performance with an average area under curve value of 0.928 for discovery cohort and 0.905 forvalidation cohort (95 % confidence interval: 0.902-0.954 and 0.845-0.965, respectively). Tandem MS and Fourier transform ion cyclotron resonance MS with ultrahigh resolution identified seven specific metabolites that were significantly associated with DR in fully adjusted models. Ofthese metabolites, dihydrobiopterin, phosphoserine, N-arachidonoylglycine, and 3-methylhistamine levels in plasma were first reported to show the associations. This work advances the design of metabolic analysis for DR and holds the potential to promise as an efficient tool for clinical management of DR.

Qualitative and quantitative studies on chemical constituents of Ling-gui-zhu-gan decoction: In vitro and in vivo.

Ling-gui-zhu-gan decoction has significant therapeutic effects in the treatment of diseases related to phlegm and fluid retention. In this study, we aimed to qualitatively characterize the chemical constituents of Ling-gui-zhu-gan decoction in vitro and in vivo by HPLC coupled to Fourier transform ion cyclotron resonance MS, and quantitively determine the contents of typical chemical constituents by HPLC method. As a result, a total of 75 chemical constituents were discovered including 37 flavonoids and their glycosides, 20 saponins, 9 sterols, 3 organic acids and their derivatives, 3 lactones, 2 coumarins, and 1 alcohol. Among them, 17 chemical constituents were specifically identified. Subsequently, an HPLC method was established for simultaneous determination of seven chemical constituents. Finally, a total of 40 prototype components were initially detected by HPLC-MS method in the biological samples of rats after their water extract was orally administrated. Among them, 29, 27, 12, and 32 prototype components were detected in plasma, bile, urine, and feces, respectively. Moreover, 34 metabolites, including 16 phase II metabolites, were detected for the first time. In conclusion, this study lays the foundation for the identification of chemical components in vitro and in vivo and the elucidation of the potential pharmacodynamic components of Ling-gui-zhu-gan decoction.

Large Individual Ion FTICR Measurements from the Mid-1990s Using Reactions for Charge Determination Mass Spectrometry.

This article discusses from a personal and present-day perspective the first studies of large highly charged individual molecular ions that were conducted using electrospray ionization with Fourier transform ion cyclotron resonance MS in the mid-1990s. These studies are distinguished from Current Charge Detection Mass Spectrometry (CDMS) primarily by their use of individual ion charge state changes due to reactions for accurate charge determination. This work describes the key differences in technologies and methods with present CDMS and the likely implications of these differences. I comment on surprising individual ion behavior observed in some measurements involving increases in charge state, as well as their possible basis, and also briefly discuss the potential utility of the reaction-based mass measurement approach used in the context of what might more globally be referred to as "Charge Determination Mass Spectrometry".

Constraints on isomers of dissolved organic matter in aquatic environments: Insights from ion mobility mass spectrometry

Elucidating the chemical structure of dissolved organic matter (DOM) is key to understanding this large yet enigmatic carbon pool. Over the last two decades much progress has been made in assigning exceptionally accurate molecular formulas of DOM owing to the application of ultrahigh resolution mass spectrometry, but little is known about the number of isomers in each molecular formula, a question essentially related to the total number of organic molecules in DOM. Such information is critical for a further understanding of the formation and long-term stability of refractory DOM in the ocean. In this work, we used ion mobility quadrupole time of flight liquid chromatography tandem mass spectrometry (IM Q-TOF LC/MS/MS) to analyze DOM samples collected in different aquatic environments including south Texas rivers, Gulf of Mexico, and South China Sea. Our data showed that generally less than 23% of all detected DOM formulas, which shared a small fraction of common molecules (ca. 12%) detected by direct-infusion Fourier transform ion cyclotron resonance MS, contained structurally distinct isomers (represented by “isomer clusters”). In addition, isomer diversity, in terms of how different the structures are, decreased with degradation in both natural and incubation samples. Specifically, the number of structurally distinct isomers was lower at river mouth and open ocean than coastal waters, where organic matter tends to be fresher due to high primary production; and with depth in water column, isomer diversity of DOM also decreased. Results from a set of incubation experiments also showed that the percentages of riverine DOM molecules that have multiple isomer clusters decreased with time, suggesting that biodegradation decreases the diversity of molecules from an isomer perspective. Overall, these results suggested that isomers, at least in a certain fraction of DOM, are highly constrained, and that degradation decreases its isomeric diversity.

Study on the Pretreatment Process and Removal Rules of Sulfur-Containing Compounds for Medium- and Low-Temperature Coal Tar.

The heteroatoms (sulfur and nitrogen) and metals (ferrum and calcium) in coal tar can easily cause the corrosion of hydrogenation equipment, catalyst poisoning, and environmental pollution. These should be removed before coal tar is hydrogenated. In this study, with the acid refining method, the effects of three polyether demulsifiers (i.e., PD1, PD2, and PD3), polyamine carboxylate demetallizers (i.e., PCD1, PCD2, and PCD3), and separation temperature on the removal of ferrum, calcium, sulfur, and nitrogen in medium- and low-temperature coal tar were determined. PD2 was selected, and the added amount was 200 μg·g–1. When the PD2 demulsifier was added alone or PD2 demulsifier with various demetallization agents was added, heteroatoms in coal tar could be effectively removed. For the experiments and analysis, the pretreatment conditions of coal tar were as follows: the addition amount of the PD2 demulsifier was 200 μg·g–1, the addition amount of the PCD3-type demetallization agent was 400 μg·g–1, and the stirring temperature was 80 °C. Before and after pretreatment, the methods of inductively coupled plasma–atomic emission spectrometer, gas chromatography–mass spectrometry (MS), and Fourier transform-ion cyclotron resonance MS were used in the present study to explore and analyze the distribution, occurrence form, and removal law of sulfur in coal tar. As revealed from the results, sulfur compounds in coal tar <360 °C fraction (light coal tar fraction, LF) before being pretreated had a lower content, which existed as benzothiophene and dibenzothiophene largely. Sulfur compounds S1 and S2 achieved the maximum relative abundance in >360 °C fraction (heavy coal tar fraction, HF). After the compounds were pretreated, the sulfur removal rate reached 40.0% in LF, and the sulfur compounds were primarily removed. For HF, the sulfur removal rate reached 20.1%. In addition, S1 compounds within the dibenzothiophene derivatives exhibiting more side chains and a larger condensation degree were basically removed. S2 compounds, mainly linked to several quinolines or more aromatic rings and thioether-aliphatic amine sulfur compounds exhibiting small molecular weight and simple structures, were relatively easy to remove. The SO class (e.g., the sulfones and thiophene-ketone group) was more difficult to remove.

Mapping glucose metabolites in the normal bovine lens: Evaluation and optimisation of a matrix-assisted laser desorption/ionisation imaging mass spectrometry method.

The spatial resolution of microdissection-based analytical methods to detect ocular lens glucose uptake, transport and metabolism are poor, whereas the multiplexing capability of fluorescence microscopy-based approaches to simultaneously detect multiple glucose metabolites is limited in comparison with mass spectrometry-based methods. To better understand lens glucose transport and metabolism, a more highly spatially resolved technique that maintains the fragile ocular lens tissue is required. In this study, a sample preparation method for matrix-assisted laser desorption/ionisation imaging mass spectrometry (MALDI IMS) analysis of ocular lens glucose uptake and metabolism has been evaluated and optimised. Matrix choice, tissue preparation and normalisation strategy were determined using negative ion mode MALDI-Fourier transform-ion cyclotron resonance MS of bovine lens tissue and validation performed using gas chromatography-MS. An internal standard was applied concurrently with N-(1-naphthyl)ethylenediamine dihydrochloride (NEDC) matrix to limit cracking of the fresh frozen lens tissue sections. MALDI IMS data were collected at a variety of spatial resolutions to detect both endogenous lens metabolites and stable isotopically labelled glucose introduced by ex vivo lens culture. Using this approach, initial steps in important metabolic processes that are linked to diabetic cataract formation were spatially mapped in the bovine lens. In the future, this method can be applied to study the dynamics of glucose uptake, transport and metabolic flux to aid in the study of diabetic lens cataract pathophysiology.

An approach for broad molecular imaging of the root-soil interface via indirect matrix-assisted laser desorption/ionization mass spectrometry

Understanding rhizospheric processes is limited by the need for imaging complex molecular transformations at relevant spatial scales within the root soil continuum. Here, we demonstrate a method to enable this analysis by first extracting organic compounds from the rhizosphere onto a PVDF membrane while maintaining their 2D distribution and then imaging the distribution of chemical compounds using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). This approach permitted us to visualize and identify compounds on the root surface and presumed root exudates in the rhizosphere. Within a 1.8 cm × 0.6 cm sampling area of a switchgrass rhizosphere, we could observe at least four chemically distinct zones. Using high performance Fourier transform ion cyclotron resonance MS, we were able to accurately annotate numerous molecules co-localized to each of these zones.

Comprehensive Compositional and Structural Comparison of Coal and Petroleum Asphaltenes Based on Extrography Fractionation Coupled with Fourier Transform Ion Cyclotron Resonance MS and MS/MS Analysis

A recently developed extrography separation method fractionates petroleum asphaltenes based on their ionization efficiency, which correlates with polarity, aggregation tendency, and asphaltene stru...

Water-Soluble Organic Composition of the Arctic Sea Surface Microlayer and Association with Ice Nucleation Ability.

Organic matter in the sea surface microlayer (SML) may be transferred to the atmosphere as sea spray and hence influence the composition and properties of marine aerosol. Recent work has demonstrated that the SML contains material capable of heterogeneously nucleating ice, but the nature of this material remains largely unknown. Water-soluble organic matter was extracted from SML and underlying seawater from the Arctic and analyzed using a combination of mass spectrometric approaches. High performance liquid chromatography-ion trap mass spectrometry (LC-IT-MS), and Fourier transform ion cyclotron resonance MS (FT-ICR-MS), showed seawater extracts to be compositionally similar across all stations, whereas microlayer extracts had a different and more variable composition. LC-IT-MS demonstrated the enrichment of particular ions in the microlayer. Ice nucleation ability (defined as the median droplet freezing temperature) appeared to be related to the relative abundances of some ions, although the extracts themselves did not retain this property. Molecular formulas were assigned using LC-quadrupole time-of-flight MS (LC-TOF-MS2) and FT-ICR-MS. The ice nucleation tracer ions were associated with elevated biogenic trace gases, and were also observed in atmospheric aerosol collected during the summer, but not early spring suggesting a biogenic source of ice nuclei in the Arctic microlayer.

Accurate Identification of Unknown and Known Metabolic Mixture Components by Combining 3D NMR with Fourier Transform Ion Cyclotron Resonance Tandem Mass Spectrometry.

Metabolite identification in metabolomics samples is a key step that critically impacts downstream analysis. We recently introduced the SUMMIT NMR/mass spectrometry (MS) hybrid approach for the identification of the molecular structure of unknown metabolites based on the combination of NMR, MS, and combinatorial cheminformatics. Here, we demonstrate the feasibility of the approach for an untargeted analysis of both a model mixture and E. coli cell lysate based on 2D/3D NMR experiments in combination with Fourier transform ion cyclotron resonance MS and MS/MS data. For 19 of the 25 model metabolites, SUMMIT yielded complete structures that matched those in the mixture independent of database information. Of those, seven top-ranked structures matched those in the mixture, and four of those were further validated by positive ion MS/MS. For five metabolites, not part of the 19 metabolites, correct molecular structural motifs could be identified. For E. coli, SUMMIT MS/NMR identified 20 previously known metabolites with three or more 1H spins independent of database information. Moreover, for 15 unknown metabolites, molecular structural fragments were determined consistent with their spin systems and chemical shifts. By providing structural information for entire metabolites or molecular fragments, SUMMIT MS/NMR greatly assists the targeted or untargeted analysis of complex mixtures of unknown compounds.

Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health

Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex4HexNAc2 to Hex7HexNAc6dHex1Neu5Ac4, as well as that of 39 glycosylation traits derived thereof. Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health.

DNA Interactions Probed by Hydrogen-Deuterium Exchange (HDX) Fourier Transform Ion Cyclotron Resonance Mass Spectrometry Confirm External Binding Sites on the Minichromosomal Maintenance (MCM) Helicase

The archaeal minichromosomal maintenance (MCM) helicase from Sulfolobus solfataricus (SsoMCM) is a model for understanding structural and mechanistic aspects of DNA unwinding. Although interactions of the encircled DNA strand within the central channel provide an accepted mode for translocation, interactions with the excluded strand on the exterior surface have mostly been ignored with regard to DNA unwinding. We have previously proposed an extension of the traditional steric exclusion model of unwinding to also include significant contributions with the excluded strand during unwinding, termed steric exclusion and wrapping (SEW). The SEW model hypothesizes that the displaced single strand tracks along paths on the exterior surface of hexameric helicases to protect single-stranded DNA (ssDNA) and stabilize the complex in a forward unwinding mode. Using hydrogen/deuterium exchange monitored by Fourier transform ion cyclotron resonance MS, we have probed the binding sites for ssDNA, using multiple substrates targeting both the encircled and excluded strand interactions. In each experiment, we have obtained >98.7% sequence coverage of SsoMCM from >650 peptides (5-30 residues in length) and are able to identify interacting residues on both the interior and exterior of SsoMCM. Based on identified contacts, positively charged residues within the external waist region were mutated and shown to generally lower DNA unwinding without negatively affecting the ATP hydrolysis. The combined data globally identify binding sites for ssDNA during SsoMCM unwinding as well as validating the importance of the SEW model for hexameric helicase unwinding.

Exemplifying the Screening Power of Mass Spectrometry Imaging over Label-Based Technologies for Simultaneous Monitoring of Drug and Metabolite Distributions in Tissue Sections

Mass spectrometry imaging (MSI) provides pharmaceutical researchers with a suite of technologies to screen and assess compound distributions and relative abundances directly from tissue sections and offer insight into drug discovery–applicable queries such as blood-brain barrier access, tumor penetration/retention, and compound toxicity related to drug retention in specific organs/cell types. Label-free MSI offers advantages over label-based assays, such as quantitative whole-body autoradiography (QWBA), in the ability to simultaneously differentiate and monitor both drug and drug metabolites. Such discrimination is not possible by label-based assays if a drug metabolite still contains the radiolabel. Here, we present data exemplifying the advantages of MSI analysis. Data of the distribution of AZD2820, a therapeutic cyclic peptide, are related to corresponding QWBA data. Distribution of AZD2820 and two metabolites is achieved by MSI, which [14C]AZD2820 QWBA fails to differentiate. Furthermore, the high mass-resolving power of Fourier transform ion cyclotron resonance MS is used to separate closely associated ions.

On the use of electron capture rate constants to describe electron capture dissociation mass spectrometry of peptides.

Electron capture dissociation (ECD) tandem mass spectrometry (MS/MS) is a powerful analytical tool for peptide and protein structure analysis. The product ion abundance (PIA) distribution in ECD MS/MS is known to vary as a function of electron irradiation period. This variation complicates the development of a method of peptide identification by correlation of ECD MS/MS data with experimental and theoretical mass spectra. Here, we first develop a kinetic model to describe primary electron capture by peptide dications leading to product ion formation and secondary electron capture resulting in product ion neutralization. We apply the developed kinetic model to calculate product ion formation rate constants and electron capture rate constants of product ions from ECD mass spectra acquired using various electron irradiation periods. Contrary to ECD PIA distributions, the product ion formation rate constants are shown to be independent of electron irradiation period and, thus, may be employed to characterize ECD product ion formation more universally. The electron capture rate constants of product ions in ECD Fourier transform ion cyclotron resonance MS were found to correlate (with a correlation factor, R(2), of ca 0.9) with ion mobility cross sections of product ions in electron transfer dissociation. Finally, we demonstrate that the electron irradiation period influences the ratio of radical and even-electron c and z product ions.

Matrix coating assisted by an electric field (MCAEF) for enhanced tissue imaging by MALDI-MS.

A novel technique, termed matrix coating assisted by an electric field (MCAEF), for enhancing tissue imaging by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was developed in this study. In this technique a static and uniform electric field is applied to sliced tissue sections during matrix spray-coating, resulting in the enrichment of positively or negatively chargeable analytes in the MALDI matrix layer. Experimental results show that MCAEF not only increased the sensitivity of lipid and protein detection across the board in the subsequent MALDI-MS analyses, but also resulted in successful imaging of a larger number of analytes. MALDI imaging enhancement with MCAEF was observed for various tissues (rat liver, rat brain, and porcine adrenal gland) and with different MALDI matrices (e.g., quercetin, 2-mercaptobenzothiazole, dithranol, 9-aminoacridine, and sinapinic acid) and the sensitivity increases were independent of the solvent compositions and pH values of the matrix solutions. Taking rat brain as an example, MCAEF led to the on-tissue detection and imaging of 648 identified lipids by combining positive and negative ion detection by MALDI-Fourier transform ion cyclotron resonance MS and with quercetin as the matrix, as compared to only 344 lipids without MCAEF. For protein imaging, up to 232 protein signals were successfully detected in rat brain tissue sections by MALDI-time-of-flight MS within a mass range of 3500 to 37 000 Da, as compared to 119 without MCAEF. MCAEF also enabled the detection of higher molecular-weight proteins. These results demonstrate the advantages of MCAEF for overall performance improvements in MALDI imaging and we believe that this technique has the potential to become a standard practice for MALDI tissue imaging.

CuO-oxidized dissolved organic matter (DOM) investigated with comprehensive two dimensional gas chromatography-time of flight-mass spectrometry (GC × GC-TOF-MS)

GC×GC-TOF-MS was applied to investigate the molecular diversity of CuO-oxidized, trimethylsilyl (TMS) derivatized DOM isolated from Lake Tahoe’s main tributary, the Upper Truckee River (UTR), California. Many (4789) chromatographic peaks were resolved in a single sample at a signal/noise (S/N) ratio of 10 or greater and>300 (>6%) were assigned discrete structures. Mapping the two dimensional elution time space revealed 8 homologous series defined by successive CH2 additions. Series of other compounds related to one another by OCH3 and OH substitutions were also assigned. Elucidation of the retention time (RT) displacements affected by these molecular transformations guided the discovery of novel compounds as well as those that had previously escaped detection within DOM, including a suite of lignin-derived cyclobutane photodimers. Analysis of RT shifts among sets of benzene polycarboxylic acid (BPCA) isomers revealed the second chromatographic dimension to strongly retain sterically strained isomers, providing a basis for assigning groups of isomers that were otherwise unassignable. The heightened chromatographic resolution also revealed a substantial and hitherto analytically inaccessible diversity of isomers, which appear to be uniquely resolved with the method, but which would fall outside the analytical window of other high resolution MS methods, such as Fourier transform ion cyclotron resonance MS (FT-ICR-MS). The study likely represents the most comprehensive compound-specific elucidation of any natural OM (NOM) sample to date.

Altering O-Linked β-N-Acetylglucosamine Cycling Disrupts Mitochondrial Function

Mitochondrial impairment is commonly found in many diseases such as diabetes, cancer, and Alzheimer disease. We demonstrate that the enzymes responsible for the addition or removal of the O-GlcNAc modification, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), respectively, are critical regulators of mitochondrial function. Using a SILAC (stable isotope labeling of amino acids in cell culture)-based proteomics screen, we quantified the changes in mitochondrial protein expression in OGT- and OGA-overexpressing cells. Strikingly, overexpression of OGT or OGA showed significant decreases in mitochondria-localized proteins involved in the respiratory chain and the tricarboxylic acid cycle. Furthermore, mitochondrial morphology was altered in these cells. Both cellular respiration and glycolysis were reduced in OGT/OGA-overexpressing cells. These data demonstrate that alterations in O-GlcNAc cycling profoundly affect energy and metabolite production.

The Crystal Structure of Shiga Toxin Type 2 with Bound Disaccharide Guides the Design of a Heterobifunctional Toxin Inhibitor

Shiga toxin type 2 (Stx2a) is clinically most closely associated with enterohemorrhagic E. coli O157:H7-mediated hemorrhagic colitis that sometimes progresses to hemolytic-uremic syndrome. The ability to express the toxin has been acquired by other Escherichia coli strains, and outbreaks of food poisoning have caused significant mortality rates as, for example, in the 2011 outbreak in northern Germany. Stx2a, an AB5 toxin, gains entry into human cells via the glycosphingolipid receptor Gb3. We have determined the first crystal structure of a disaccharide analog of Gb3 bound to the B5 pentamer of Stx2a holotoxin. In this Gb3 analog,-GalNAc replaces the terminal-Gal residue. This co-crystal structure confirms previous inferences that two of the primary binding sites identified in theB5 pentamer of Stx1 are also functional in Stx2a. This knowledge provides a rationale for the synthesis and evaluation of heterobifunctional antagonists for E. coli toxins that target Stx2a. Incorporation of GalNAc Gb3 trisaccharide in a heterobifunctional ligand with an attached pyruvate acetal, a ligand for human amyloid P component, and conjugation to poly[acrylamide-co-(3-azidopropylmethacrylamide)] produced a polymer that neutralized Stx2a in a mouse model of Shigatoxemia.

Quantification of intact covalently metal labeled proteins using ESI-MS/MS

Mass spectrometric methods matured from the successful qualitative characterization of proteins in complex mixtures into methods for quantitative proteomics often based on chemical tags with stable isotope labeling. In the study presented here, we extended the application of lanthanide-ion-based tags from the quantification using inductively coupled plasma-MS into the quantification of labeled intact proteins using electrospray ionization (ESI)-MS and ESI-MS/MS. We applied the metal chelate tag MeCAT-iodoacetamide (IA) (1,4,7,10-tetraazacyclododecane N,N',N″,N″ '-tetra acetic acid with a IA reactive site). Labeled proteins were separated using C3-reversed phase-high-performance liquid chromatography interfaced to ESI-MS. We could prove that even large proteins were completely labeled at all available cysteine residues using MeCAT-IA with only a small excess of reagent. Fragmentation of labeled proteins either using infrared multiphoton dissociation in Fourier transform ion cyclotron resonance-MS or higher-energy collision dissociation with an Orbitrap gave characteristic fragments. We used these fragments to quantify several intact proteins avoiding digestion. To demonstrate the applicability, human serum albumin was quantified in blood serum. The high-performance liquid chromatography/ESI-MS/MS quantification data were validated using inductively coupled plasma-MS. Because the metal within the tag may be any of the lanthanides, multiplexing capabilities are inherent.

Characterization of polyisobutylene succinic anhydride chemistries using mass spectrometry

AbstractRepresentative Polyisobutylene succinic anhydride (PIBSA) samples have been studied by different MS techniques including Electrospray Ionization Fourier Transform Ion Cyclotron Resonance MS (ESI‐FTMS) (positive and negative modes), Atmospheric Pressure Chemical Ionization Fourier Transform Ion Cyclotron Resonance MS (negative mode), and Matrix‐Assisted Laser Desorption/Ionization Time‐of‐Flight (positive and negative modes). Negative ion ESI‐FTMS produces the best results. Differences between “mono‐succan” and “di‐succan” content can readily be observed. The source of the PIBSA (PIBSA‐I and PIBSA‐II processes) can be easily distinguished and the formation of methyl esters and amide derivatives can provide complementary data. The experiments have demonstrated the capabilities of mass spectrometry to detect and characterize such polymers samples. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012