ESI-TOF MS Research Articles

Penilumamide, a novel lumazine peptide isolated from the marine-derived fungus, Penicillium sp. CNL-338.

A novel lumazine peptide, penilumamide (1), was isolated from the fermentation broth of a marine-derived fungal strain, identified as Penicillium sp. (strain CNL-338) and the structure of the new metabolite was determined by analysis of ESI-TOF MS data combined with 1D and 2D NMR experiments.

A combined XAFS, ESI TOF-MS and LIBD study on the formation of polynuclear Zr(IV), Th(IV) and Pu(IV) species

The long term radiotoxicity of spent nuclear fuel disposed of in deep underground repositories after discharge from nuclear power reactors is determined by actinide elements, mainly plutonium. Water intrusion into the repository might cause container corrosion and leaching of the waste matrices, leading to the release of Pu and other actinides into the geological environment. Performance assessment for a future nuclear waste repository requires detailed knowledge on actinide aqueous chemistry in the aquifer surrounding the disposal site. Tetravalent actinides exhibit a strong tendency towards hydrolysis and subsequent polymerization and/or colloid formation. These species provide a potential pathway for migration of actinides away from the repository. Therefore, it is of fundamental interest to study their generation and properties in-situ. To this end, X-ray Absorption Fine Structure Spectroscopy (XAFS) at the INE-Beamline for actinide research at ANKA, Electrospray Mass-Spectrometry (ESI TOF-MS) and Laser Induced Breakdown Detection (LIBD) are combined at FZK-INE in a comprehensive attempt to characterize Zr(IV) (An(IV) analogue), Th(IV) and Pu(IV) polymerization and colloid formation.

Anti-Selective Catalytic Asymmetric Nitroaldol Reaction via a Heterobimetallic Heterogeneous Catalyst

Full details of an anti-selective catalytic asymmetric nitroaldol reaction promoted by a heterobimetallic catalyst comprised of Nd(5)O(O(i)Pr)(13), an amide-based ligand, and NaHMDS (sodium hexamethyldisilazide) are described. A systematic synthesis and evaluation of amide-based ligands led to the identification of optimum ligand 1m, which provided a suitable platform for the Nd/Na heterobimetallic complex. During the catalyst preparation in THF, a heterogeneous mixture developed and centrifugation of the suspension allowed for separation of the precipitate, which contained the active catalyst and which could be stored for at least 1 month without any loss of catalytic performance. The precipitate promoted a nitroaldol (Henry) reaction for a broad range of nitroalkanes and aldehydes under heterogeneous conditions, affording the corresponding 1,2-nitroalkanol in a highly anti-selective (up to anti/syn = >40/1) and enantioselective manner (up to 98% ee). Inductively coupled plasma (ICP) and X-ray fluorescence (XRF) analyses revealed that the precipitate indeed included both neodymium and sodium, which was further supported by high-resolution ESI TOF MS spectrometry.

Metal ion binding of modified pyrimidine-base pairs in DNA duplexes

We report the synthesis and metal ion-binding properties of DNA duplexes containing 5-substituted uracil ((X)U) pairs, such as 5-bromouracil, 5-fluorouracil and 5-cyanouracil pairs. Thermal denaturation studies of these modified DNA duplexes revealed that the DNA duplexes were stabilized in the presence of Hg(II) ions in acidic and neutral solutions. On the other hand, the duplexes were stabilized in the presence of Hg(II) and Ag(I) ions. These results indicated that (X)U-Hg(II)-(X)U complex was formed in the acidic and neutral solutions, then, in the basic solutions (X)U-Ag(I)-(X)U complex as well as the (X)U-Hg(II)-(X)U complex were formed. ESI-TOF MS analysis also indicated formation of the metal ion-DNA complexes.

NACE‐ESI‐TOF MS to reveal phenolic compounds from olive oil: Introducing enriched olive oil directly inside capillary

Most CE methods for the analysis of phenols from olive oil use an aqueous electrolyte separation medium, although the importance of NACE is obvious, as this kind of CE seems to be more compatible with the hydrophobic olive oil matrix and could facilitate its direct injection. In the current work we develop a method involving SPE and NACE coupled to ESI-TOF MS. All the CE and ESI-TOF MS parameters were optimized in order to maximize the number of phenolic compounds detected and the sensitivity in their determination. Electrophoretic separation was carried out using a CE buffer system consisting of 25 mM NH(4)OAc/AcH in methanol/ACN (1/1 v/v) at an apparent pH value of 5.0. We studied in depth the effect of the nature and concentration of different electrolytes dissolved in different organic solvents and other experimental and instrumental CE variables. The results were compared with those obtained by CZE (with aqueous buffers) coupled to ESI-TOF MS; both methods offered to the analyst the chance to study phenolic compounds of different families (such as phenolic alcohols, lignans, complex phenols, flavonoids, etc.) from virgin olive oil by injecting methanolic extracts with efficient and fast CE separations. In the case of NACE method, we also studied the direct injection of the investigated matrix introducing a plug of olive oil directly into the capillary.

A selective fluorescence probe for yttrium(III) based on acylhydrazone Schiff base

Abstract A new fluorescence probe has been developed for the recognition of Y3+. The probe, 2-hydroxy-1-naphthaldehyde salicyloylhydrazone, L, with Y3+ was studied in detail. Based on the chelation reaction, a highly sensitive colorimetric and fluorescent method was developed for the determination of yttrium in THF; it can detect the trace Y3+ with naked-eye in the sun. The binding mode was investigated by UV–Vis spectra, ESI-TOF MS, IR and XRD.

Selective Binding of Glutathione Conjugates of Fatty Acid Derivatives by Plant Glutathione Transferases

Proteomic studies with Arabidopsis thaliana have revealed that the plant-specific Tau (U) class glutathione transferases (GSTs) are selectively retained by S-hexylglutathione affinity supports. Overexpression of members of the Arabidopsis GST superfamily in Escherichia coli showed that 25 of the complement of 28 GSTUs caused the aberrant accumulation of acylated glutathione thioesters in vivo, a perturbation that was not observed with other GST classes. Each GSTU caused a specific group of fatty acyl derivatives to accumulate, which varied in chain length (C(6) to C(18)), additional oxygen content (0 or 1), and desaturation (0 or 1). Thioesters bound tightly to recombinant GSTs (K(d) approximately 1 microm), explaining their accumulation. Transient expression of GSTUs in Nicotiana benthamiana followed by recovery by Strep-tag affinity chromatography allowed the respective plant ligands to be extracted and characterized. Again, each GST showed a distinct profile of recovered metabolites, notably glutathionylated oxophytodienoic acid and related oxygenated fatty acids. Similarly, the expression of the major Tau protein GSTU19 in the endogenous host Arabidopsis led to the selective binding of the glutathionylated oxophytodienoic acid-glutathione conjugate, with the enzyme able to catalyze the conjugation reaction. Additional ligands identified in planta included other fatty acid derivatives including divinyl ethers and glutathionylated chlorogenic acid. The strong and specific retention of various oxygenated fatty acids by each GSTU and the conservation in binding observed in the different hosts suggest that these proteins have selective roles in binding and conjugating these unstable metabolites in vivo.

Determination of Critical Conditions of Adsorption for Chromatography of Polymers

Liquid chromatography (LC) at critical conditions of adsorption was used to separate various poly(ethylene oxides), poly(propylene oxides) and their copolymers. For the first time, the determination of the critical conditions by means of Ultra Performance Liquid Chromatography (UPLC) coupled to Electrospray Ionization Time-of-flight Mass Spectrometry (ESI-TOF MS) is reported. In contrast to established, mostly laborious routines to find suitable chromatographic separation conditions, this coupling enables a very fast adjustment of parameters. Similar to LC Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (LC/MALDI MS) coupling, a two-dimensional analysis of homo- and copolymers regarding its functionality type and molecular weight distribution, as well as copolymer composition, can be performed simultaneously. Furthermore, there is no need for using polymer standards for the determination of critical conditions or Size Exclusion Chromatography calibration.

Complete Characterizing of Erythropoietin Glycoforms Using Capillary Zone Electrophoresis Coupled to Mass Spectrometry

Erythropoietin (EPO) plays a key role in the production of red blood cells and is currently one of the most important approved drugs. EPO is a glycoprotein of about 30kDa, where the glycosylation has an influence on its lifetime in blood, biological availability and activity. Due to this wide glycoform distribution the direct mass spectral characterization either by ESI or by MALDI failed. Here we show for the first time a method for the accurate mass determination of intact isomeric glycoforms of EPO by ESI-TOF MS, based on prior on-line separation by capillary electrophoresis.

Two new lanostane triterpenoids from Poria cocos

Two new lanostane triterpenoids, 29-hydroxypolyporenic acid C (4) and 25-hydroxypachymic acid (5), together with three known compounds, ergosta-7,22-dien-3β-ol (1), polyporenic acid C (2) and pachymic acid (3), were isolated from the 95% ethanolic extract of the sclerotium of Poria cocos (Schw.) Wolf. Their structures were determined by extensive spectroscopic analyses, including IR, UV, ESITOF–MS, HRESI–MS, 1D and 2D NMR data (1H NMR, 13C NMR, 1H–1H COSY, NOESY, HSQC and HMBC).

Nanoscale Affinity Chip Interface for Coupling Inhibition SPR Immunosensor Screening with Nano-LC TOF MS

The on-line nanoscale coupling of a surface plasmon resonance (SPR)-based inhibition biosensor immunoassay (iBIA) for the screening of low molecular weight molecules with nano-liquid-chromatography electrospray ionization time-of-flight mass spectrometry (nano-LC ESI TOF MS) for identification is described. The interface is based on a reusable recovery chip (RC) that contains a nanoscale biosorbent composed of a hydrogel layer modified with antibodies raised against the analyte featuring the unique possibility of performance characterization using the SPR biosensor. Various hydrogel chemistries were evaluated, and the standard Biacore CM5 chip showed the highest capture capacity in combination with affinity-purified polyclonal antibodies. The procedure has four stages: the samples are prepared (1) and screened using a screening chip (SC) in the iBIA (2). Suspected noncompliant samples as being noncompliant are reinjected over the RC, and the analyte is captured at subnanogram level (3). The captured analyte is released, and the eluate is analyzed with nano-LC ESI TOF MS via a loop-type interface (4). The coupling of the technologies proved effective for screening enrofloxacin, a model compound, in incurred chicken muscle samples followed by identity confirmation in suspected noncompliant samples. Ciprofloxacin, a known metabolite of enrofloxacin, was identified as well in incurred chicken samples. This demonstrates the potential of the technologies coupled by means of a RC for the rapid screening and identification of known as well as unknown compounds. Finally, we demonstrate the feasibility of combining the two biosensor chips (SC and RC) with a robust chip-based nano-LC chip TOF MS system, thus providing a robust alternative triple-chip system.

Preparation and metal ion-binding of 4-N-substituted cytosine pairs in DNA duplexes

We report the synthesis and metal ion-binding properties of DNA duplexes containing 4-N-substituted cytosine base pairs. Thermal denaturation studies of these modified DNA duplexes in the presence of various metal ions revealed that the DNA duplexes containing 4-N-carboxymethylcytosine (1) base pair(s) bound Ag (I), Ni(II), and Cu(II) ions. Moreover, ESI-TOF MS analysis of the DNA duplexes containing 1-1 base pair(s) in the presence of Cu(II) ions was consistent with one 1-1 base pair region binding one equivalent of Cu(II) ions. These results indicate that the DNA duplexes containing 1-1 base pair(s) may be useful as a new metal ion-binding motif.

Characterization and Structure Analysis of a Novel Bacteriocin, Lacticin Z, Produced byLactococcus lactisQU 14

A novel bacteriocin, lacticin Z, produced by Lactococcus lactis QU 14 isolated from a horse's intestinal tract was identified. Lacticin Z was purified through a three step procedure comprised of hydrophobic-interaction, cation-exchange chromatography, and reverse-phase HPLC. ESI-TOF MS determined the molecular mass of lacticin Z to be 5,968.9 Da. The primary structure of lacticin Z was found to consist of 53 amino acid residues without any leader sequence or signal peptide. Lacticin Z showed homology to lacticin Q from L. lactis QU 5, aureocin A53 from Staphylococcus aureus A53, and mutacin BHT-B from Streptococcus rattus strain BHT. It exhibited a nanomolar range of MICs against various Gram-positive bacteria, and the activity was completely stable up to 100 degrees C. Unlike many of other LAB bacteriocins, the stability of lacticin Z was emphasized under alkaline conditions rather than acidic conditions. All the results indicated that lacticin Z belongs to a novel type of bacteriocin.

Comparative Proteomics Analysis of Barrett Metaplasia and Esophageal Adenocarcinoma Using Two-dimensional Liquid Mass Mapping

Esophageal adenocarcinoma, currently the seventh leading cause of cancer-related death, has been associated with the presence of Barrett metaplasia. The malignant potential of Barrett metaplasia is evidenced by ultimate progression of this condition to invasive adenocarcinoma. We utilized liquid phase separation of proteins with chromatofocusing in the first dimension and nonporous reverse phase HPLC in the second dimension followed by ESI-TOF mass spectrometry to identify proteins differentially expressed in six Barrett metaplasia samples as compared with six esophageal adenocarcinoma samples; all six Barrett samples were obtained from the identical six patients from whom we obtained the esophageal adenocarcinoma tissue. Approximately 300 protein bands were detected by mass mappings, and 38 differentially expressed proteins were identified by microLC-MS/MS. The false positive rates of the peptide identifications were evaluated by reversed database searching. Among the proteins that were identified, Rho GDP dissociation inhibitor 2, alpha-enolase, Lamin A/C, and nucleoside-diphosphate kinase A were demonstrated to be up-regulated in both mRNA and protein expression in esophageal adenocarcinomas relative to Barrett metaplasia. Candidate proteins were examined at the mRNA level using high density oligonucleotide microarrays. The cellular expression patterns were verified in both esophageal adenocarcinomas and in Barrett metaplasia by immunohistochemistry. These differentially expressed proteins may have utility as useful candidate markers of esophageal adenocarcinoma.

Hydrolysis products of water treatment chemical aluminium sulfate octadecahydrate by electrospray ionization mass spectrometry

Hydrolysis and speciation of aluminium sulfate octadecahydrate Al2(SO4)3·18H2O was studied by electrospray time of flight mass spectrometry (ESI TOF MS). Several novel polymeric species were determined. Highly charged polymers, characterized by other methods, such as the Keggin cation [Al13O4(OH)24(H2O)12]7+ and the octameric aluminium hydroxide cluster [Al8(OH)14(H2O)18](SO4)5 16H2O, were found using ESI-MS as the anions [Al13O4(OH)25(SO4)4]2− and [Al8O(OH)14(SO4)5(H2O)4]2−. All the main species identified contained sulfate or hydrogen sulfate. The compositions of the determined ions mimicked those of several stable mineral forms.

Electrospray Mass Spectrometry Study of Tiopronin Monolayer-Protected Gold Nanoclusters

Electrospray ionization time-of-flight mass spectrometry (ESI-TOF MS) and gel permeation chromatography (GPC) were used to study the synthesis of a series of tiopronin monolayer-protected gold nanoclusters (MPCs) and to monitor their postsynthesis peptide ligand place-exchange reactions. All mass spectra identified the presence of cyclic gold(I)-thiolates with a strong preference for tetrameric species. During the synthesis of pre-monolayer-protected nanoclusters (pre-MPCs), esterified gold(I)-thiolate tetramers were initially observed in minor abundance (with respect to disulfide bridged tiopronin species) before dramatically increasing in abundance and precipitating from solution. After conversion of pre-MPCs to MPCs, ESI-TOF mass spectra demonstrated an overall predominance of tetrameric species with conversion from ester-terminated end groups to carboxyl-terminated end groups. Further modifications were performed through postsynthesis ligand place-exchange reactions to validate the existence of the tetramers. This work suggests that monolayer protection is accomplished by cyclized gold(I)-thiolate tetramers on the gold core surface, and/or that gold(I)-thiolates are a basic building block within the nanoparticles.

Controlled-potential electrosynthesis of glucosaminic acid from glucosamine at a gold electrode

The electrocatalytic oxidation of d-glucosamine (2-amino-2-deoxy- d-glucose) in alkaline and neutral solutions was examined using a carbon felt electrode modified with 2 nm core sized gold nanoparticles (Au 2 nm nanoparticles) and a gold plate electrode. The electrocatalytic voltammetric oxidation curves of d-glucosamine were obtained in both solutions. The voltammetric responses for the electrocatalytic oxidation at a Au 2 nm nanoparticle-modified electrode in both alkaline and neutral solutions were almost the same to those at a gold plate electrode. The oxidized product was identified to be d-glucosaminic acid (2-amino-2-deoxy- d-gluconic acid) generated by the 2-electron oxidation product of d-glucosamine by electrospray ionization time-of-flight mass spectra (ESI TOF-MS). The HPLC results also indicated that the oxidation product was d-glucosaminic acid. The controlled-potential electrolysis of d-glucosamine was performed at the Au 2 nm nanoparticle-modified carbon felt electrodes in both alkaline and neutral solutions. In the alkaline solution, at a potential of −0.2 V, d-glucosaminic acid was formed with a current efficiency of 100%. In the neutral solution, electrolysis at 0.4 V on d-glucosaminic acid was obtained with current efficiencies of 70%.

A study of the Pb(II) binding to recombinant mouse Zn 7-metallothionein 1 and its domains by ESI TOF MS

This work reports, for the first time, a complete analysis of the binding of Pb(II) ions to mammalian Zn-metallothionein (MT) and its individual domains (α and β) by means of ESI TOF MS. The use of highly pure recombinant Zn-MT and high resolution MS allowed the identification of the different heteronuclear Pb,Zn-MT and homonuclear Pb-MT complexes formed by Zn/Pb replacement in the initial Zn-MT complexes at pH 7. The results obtained showed, both for the whole Zn 7-MT and for its individual Zn 4-α and Zn 3-β fragments, a partial substitution of the Zn initially bound to give rise to the formation of both Pb,Zn-MT and Pb-MT species. In the case of the whole Zn 7-MT, the addition of seven equivalents of Pb(II) give rise to the formation of several Pb x Zn y -MT species ( x + y = 7 or 8) as well as Pb 8-MT and Pb 9-MT coexisting with the initial Zn 7-MT species. The addition of a five-fold excess of Pb(II) showed the formation of species containing a higher amount of lead(II), Pb x Zn y -MT species ( x = 8–12, y = 0–1). In both cases, the evolution with time showed the disappearance of the species containing lead and concomitantly only detection of the initial Zn-MT clusters within 48 h. This behaviour was also observed for the Zn 4-α and Zn 3-β aggregates, confirming the reversibility of the Pb 2+ displacement of Zn 2+ ions in the three forms of MT. A similar result was obtained at acidic pH, when the apo-form was the major species in solution, this therefore confirming the low affinity of MT for Pb(II), even in the absence of zinc. The results obtained were consistent with the increase of MT synthesis in response to Pb 2+ administered to rats reported in the literature, since the initial release of Zn(II) would promote the biosynthesis of MT, in agreement with the function of this ion as primary activator of the MTF-1 transcription factor; and could explain the difficulty of recovering homonuclear Pb-MT species from lead-treated organisms.

Comparison of hydrolysis products of AlCl3 ·6H2O in different concentrations by electrospray ionization time of flight mass spectrometer (ESI TOF MS)

Hydrolysis of 0.100, 0.010, and 0.001 mol L−1 aqueous solutions of AlCl3·6H2O, each at two pH values ranging from 3.27 to 7.00, was compared by electrospray ionization time-of-flight mass spectrometry. The differences between 4 h and 14 day aged solutions were also studied. Various charges and compositions for species of the same size were observed. At lower pH values, dimeric and trimeric aluminium oxo-hydroxo-complexes were the major molecular species. With higher pH levels, precipitation and formation of took place. Multiple charged signals assigned to polynuclear complexes (Al10–Al15) were found at a narrow pH area range before precipitation occurred. The diversity of different species increased strongly as a function of concentration. The pH of the solution dominated the profile even more than the concentration. All the main species were formed in 4 h.

Proteomic analysis of cold adaptation in a Siberian permafrost bacterium – Exiguobacterium sibiricum 255–15 by two‐dimensional liquid separation coupled with mass spectrometry

Bacterial cold adaptation in Exiguobacterium sibiricum 255-15 was studied on a proteomic scale using a 2-D liquid phase separation coupled with MS technology. Whole-cell lysates of E. sibiricum 255-15 grown at 4 degrees C and 25 degrees C were first fractionated according to pI by chromatofocusing (CF), and further separated based on hydrophobicity by nonporous silica RP HPLC (NPS-RP-HPLC) which was on-line coupled with an ESI-TOF MS for intact protein M(r) measurement and quantitative interlysate comparison. Mass maps were created to visualize the differences in protein expression between different growth temperatures. The differentially expressed proteins were then identified by PMF using a MALDI-TOF MS and peptide sequencing by MS/MS with a MALDI quadrupole IT TOF mass spectrometer (MALDI-QIT-TOF MS). A total of over 500 proteins were detected in this study, of which 256 were identified. Among these proteins 39 were cold acclimation proteins (Caps) that were preferentially or uniquely expressed at 4 degrees C and three were homologous cold shock proteins (Csps). The homologous Csps were found to be similarly expressed at 4 degrees C and 25 degrees C, where these three homologous Csps represent about 10% of the total soluble proteins at both 4 degrees C and 25 degrees C.