Direct analysis of oligosaccharides and alpha hydroxy acids in fruits using electrosonic spray ionization mass spectrometry

Electrosonic spray ionization (ESSI) is a derivative technique of electrospray ionization (ESI) for mass spectrometry (MS) in which droplets are charged in the course of sonic spray. In this study, we applied ESSI MS to direct analysis of oligosaccharides and alpha hydroxy acids (AHAs) in fruits. The components were extracted from fruit fleshes by a feasible method prior to ESSI MS analysis, but the fruit juices were analyzed without further pretreatment. The results demonstrate that mainly alkali metal adducts of oligosaccharides are favorably produced in positive ion mode, while deprotonated AHAs and oligosaccharides are produced in negative ion mode. Compared with mass spectra obtained using electrospray droplet impact/secondary ion mass spectrometry (EDI/SIMS), mass spectra using ESSI make the identification of oligosaccharides more straightforward in positive ion mode than in negative ion mode.     

高分辨率电喷雾飞行时间质谱测定丹参脂溶性组分

丹参 [1] (Salvia miltiorrhiza Bage)为唇形科鼠尾草属多年生草本植物 ,临床上用于治疗妇科病、冠心病、缺血性中风及动脉粥样硬化等症 .近年来 ,飞行时间质谱 (TOF- MS)的分辨率优于 1 0 4 [2 ] ,尤其是再配合 ESI/MAL DI/APCI及新型脉冲电喷雾 [3 ]等离子源 ,使之成为当今最     

Study of cyclyzation of o-(1-Alkynyl)- and o-(1,3-Butadiynyl)aryltriazenes under the action of acids

The investigation of Richter cyclization of o-(1-alkynyl)- and o-(1,3-butadiynyl)arenediazonium salts generated by acid cleavage of triazenes was carried out by the methods of NMR and ESI MS (Electrospray Ionization Mass Spectrometry). The effect was shown of the substituents at the carbon atom of the triple bond and in the aromatic ring, of the solvent, temperature, and the reagents ratio on the rate of the cyclization and the yield of its products, 4-bromo-3-alkyl- and 4-bromo-3-ethynylcinnolines; the conditions of their synthesis were optimized. The possibility to use 4-bromo-3-ethynylcinnolines as substrates of electrophilic cyclization was demonstrated.     

Internal energy distributions in desorption electrospray ionization (DESI)

The internal energy distributions of typical ions generated by desorption electrospray ionization (DESI) were measured using the "survival yield" method, and compared with corresponding data for electrospray ionization (ESI) and electrosonic spray ionization (ESSI). The results show that the three ionization methods produce populations of ions having internal energy distributions of similar shapes and mean values (1.7-1.9 eV) suggesting similar phenomena, at least in the later stages of the process leading from solvated droplets to gas-phase ions. These data on energetics are consistent with the view that DESI involves "droplet pick-up" (liquid-liquid extraction) followed by ESI-like desolvation and gas-phase ion formation. The effects of various experimental parameters on the degree of fragmentation of p-methoxy-benzylpyridinium ions were compared between DESI and ESSI. The results show similar trends in the survival yields as a function of the nebulizing gas pressure, solvent flow rate, and distance from the sprayer tip to the MS inlet. These observations are consistent with the mechanism noted above and they also enable the user to exercise control over the energetics of the DESI ionization process, through manipulation of external and internal ion source parameters.     

Determination of linear alkylbenzenesulfonates in communal wastewater by means of solid phase microextraction coupled with API-MS and HPLC-FLD

The use of Solid Phase Microextraction (SPME) for the qualitative and quantitative determination of Linear Alkylbenzenesulfonates (LAS) in waste water samples was investigated. A Carbowax/Templated Resin (CW/¶TPR) coated fiber was directly immersed into influent and effluent samples of a sewage treatment plant (STP). Extraction conditions such as time, pH, ion strength were investigated. The extracted LAS were desorbed with a solvent in a specially designed SPME-LC interface for analysis with HPLC-FLD and Electrospray Ionization Mass Spectrometry (ESI-MS). The combination of SPME with ESI-MS proved to be an alternative technique for the LAS determination in waste water. Linear ranges of the external calibration were found from 0.5–100 ng/mL, with detection limits of 0.5 ng/mL for each individual LAS homologue. The reproducibility of the method is 15% (relative standard deviation).     

Mechanism of the Anionic Ring-Opening Oligomerization of Propylene Carbonate Initiated by the tert-Butylphenol/KHCO3 System

The ring-opening oligomerization reaction of propylene carbonate in the presence of the tert-butylphenol/KHCO₃ initiating system was studied by means of Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) and Electrospray Ionization Time of Flight Mass Spectrometry (ESI-TOF MS). According to the MS spectra obtained, different series of peaks were identified. The MS spectra clearly showed that besides the chain-extension reaction yielding oligomers with all propylene oxide units, the formation of oligomers containing carbonate linkages in the chain, and condensation reaction between the latter two also took place. The structure of the oligomers carrying carbonate linkages was determined by the post-source decay (PSD) MALDI-TOF MS/MS method. Based on the MS results, a mechanism for the oligomerization reaction is proposed.     

Electrosonic spray ionization--an ideal interface for high-flow liquid chromatography applications

Electrosonic spray ionization (ESSI) has been studied as an interface between high-performance liquid chromatography (HPLC) and mass spectrometry (MS), using sample flow rates up to 3.0 ml min⁻¹. This ionization interface was compared with pneumatically assisted electrospray ionization (ESI) using mass spectrometry for detection. For experiments that did not involve direct comparison of different flow rates, the ESI experiments were performed using post column splitting to work at optimal conditions. ESSI allows the interfacing of conventional or high-resolution liquid chromatography (LC) methods to mass spectrometry without post column splitting. High sample flow rates could be handled without a significant loss of signal intensity using a nebulization gas flow rate of 5.5 L min⁻¹. Since ESI needs to be operated with lower sample flow rates, it is limited to micro/nano LC systems, or post column splitting must be used. In particular, nano LC systems have to be treated with great care and require constant maintenance. When using post-column splitting, the increased diffusion can become a problem especially when using systems with very small void volumes. In all experiments ESSI showed better signal intensities than a commercially available, pneumatically assisted ESI source. ESSI does not require heating of the nebulizer gas, which should help to preserve the original structure of thermally unstable molecules. Therefore, ESSI is presented as an alternative to the commercially available heated ESI sources of AB SCIEX, Thermo Fischer, Agilent and Waters. The observed LC-ESSI-MS ion chromatograms are shown to be very stable even when using flow rates higher than 1.0 ml min⁻¹, which could be very suitable for ultra high performance LC, where sample flow rates up to 2.0 mL min⁻¹ with backpressures up to 1200 bar are used. Also, a difference in the relative intensities of singly and doubly protonated peptide monomers and dimers was observed between the two ionization methods. The coefficients of determination for the calibration of instrument response for Val–Tyr–Val and Met-Enkephalin showed excellent linearity over a wide concentration range (0.1–100 μM), while ESI results were only linear over a much smaller range (0.1–20 μM). The observed behavior is thought to be caused by insufficient ionization efficiency of solutions above ∼20 μM by ESI, exemplifying the robustness of ESSI as an interface between LC and MS.     

Two-step on-particle ionization/enrichment via a washing- and separation-free approach: multifunctional TiO2 nanoparticles as desalting, accelerating, and affinity probes for microwave-assisted tryptic digestion of phosphoproteins in ESI-MS and MALDI-MS: comparison with microscale TiO2

We introduce a simplified sample preparation method using bare TiO₂ nanoparticles (NPs) to serve as multifunctional nanoprobes (desalting, accelerating, and affinity probes) for effective enrichment of phosphopeptides from microwave-assisted tryptic digestion of phosphoproteins (α-casein, β-casein and milk) in Electrospray Ionization Mass Spectrometry (ESI-MS) and Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS). The results demonstrate that TiO₂ NPs can effectively enrich and accelerate the digestion reactions of phosphoproteins in aqueous solutions and also from complex real samples. After the microwave experiments, we directly injected the resulting solutions into the ESI-MS and MALDI-MS systems for analysis, and excellent sensitivity was achieved without the need for any washing procedure or separation process. The reasons are attributed to the high binding affinity and selectivity of TiO₂ NPs toward phosphopeptides. Thus, phosphopeptides can be adsorbed onto the TiO₂ NP surface. The digested or partially digested phosphoproteins can be concentrated onto the TiO₂ NP surface. This results in the effective or complete digestion of phosphoproteins in a short period of time (45 s). In addition, high sensitivity and sequence coverage of phosphopeptide can be obtained using TiO₂ NPs as microwave absorbers and affinity probes in MALDI-MS and ESI-MS. This is due to the photocatalytic nature of the TiO₂ NPs because the absorption of microwave radiation that can accelerate the activation of trypsin for efficient digestion of phosphoproteins and enhances the ionization of phosphopeptides. The lowest concentrations detected for ESI-MS and MALDI-MS were 0.1 μM and 10 fmol, respectively, for α-casein. Comparing the two-step approach of TiO₂ NPs with microscale TiO₂ particles, the microscale TiO₂ particles shows no effect on the microwave-assisted tryptic digestion of phosphoproteins. The current approach offers multiple advantages, such as great simplicity, high sensitivity and selectivity, straightforward and separation/washing-free technique for phosphorpeptide enrichment analysis.     

Accelerated reactions of amines with carbon dioxide driven by superacid at the microdroplet interface

Microdroplets display distinctive interfacial chemistry, manifested as accelerated reactions relative to those observed for the same reagents in bulk. Carbon dioxide undergoes C–N bond formation reactions with amines at the interface of droplets to form carbamic acids. Electrospray ionization mass spectrometry displays the reaction products in the form of the protonated and deprotonated carbamic acid. Electrosonic spray ionization (ESSI) utilizing carbon dioxide as nebulization gas, confines reaction to the gas–liquid interface where it proceeds much faster than in the bulk. Intriguingly, trace amounts of water accelerate the reaction, presumably by formation of superacid or superbase at the water interface. The suggested mechanism of protonation of CO₂ followed by nucleophilic attack by the amine is analogous to that previously advanced for imidazole formation from carboxylic acids and diamines.

Microdroplets display distinctive interfacial chemistry, manifested as accelerated reactions relative to those observed for the same reagents in bulk.     

Macromolecule mass spectrometry: Citation mining of user documents

Identifying research users, applications, and impact is important for research performers, managers, evaluators, and sponsors. Identification of the user audience and the research impact is complex and time consuming due to the many indirect pathways through which fundamental research can impact applications. This paper identified the literature pathways through which two highly-cited papers of 2002 Chemistry Nobel Laureates Fenn and Tanaka impacted research, technology development, and applications. Citation Mining, an integration of citation bibliometrics and text mining, was applied to the >1600 first generation Science Citation Index (SCI) citing papers to Fenn's 1989 Science paper on Electrospray Ionization for Mass Spectrometry, and to the >400 first generation SCI citing papers to Tanaka's 1988 Rapid Communications in Mass Spectrometry paper on Laser Ionization Time-of-Flight Mass Spectrometry. Bibliometrics was performed on the citing papers to profile the user characteristics. Text mining was performed on the citing papers to identify the technical areas impacted by the research, and the relationships among these technical areas.     

Zinc-induced conformational changes in the DNA-binding domain of the vitamin D receptor determined by electrospray ionization mass spectrometry

Electrospray ionization mass Spectrometry (ESI-MS) was used to measure conformational changes within the DNA-binding domain of the vitamin D receptor (VDR DBD) upon binding zinc (Zn²⁺). As increasing concentrations of Zn²⁺ were added to the VDR DBD, a gradual shift in the mass envelope to lower charge states was observed in the multiply charged spectrum. The shift in the charge states was correlated to changes observed in the far-ultraviolet circular dichroic (far-UV CD) spectrum of the protein as it was titrated with Zn²⁺. Both the multiply charged ESI and far-UV CD spectra of the Zn²⁺-titrated protein show that the binding of the first Zn²⁺ ion to the protein results in very little conformational change in the protein. The binding of a second Zn²⁺ ion resulted in a significant alteration in the structure of the protein as indicated by changes in both the multiply charged ESI and far-UV CD spectra. Much smaller changes were seen within the multiply charged ESI or far-UV CD spectra upon increasing the Zn²⁺ concentration beyond 2 mol/mol of protein. The results presented indicate that ESI-MS in combination with CD is a powerful method to measure gross conformational changes induced by the binding of metals to metalloproteins.     

Electron impact ionization of CCl4 and SF6 embedded in superfluid helium droplets

Electron impact ionization of helium nano-droplets containing several 10⁴ He atoms and doped with CCl₄ or SF₆ molecules is studied with high-mass resolution. The mass spectra show significant clustering of CCl₄ molecules, less so for SF₆ under our experimental conditions. Positive ion efficiency curves as a function of electron energy indicate complete immersion of the molecules inside the helium droplets in both cases. For CCl₄ we observe the molecular parent cation CCl₄⁺ that preferentially is formed via Penning ionization upon collisions with He*. In contrast, no parent cation SF₆⁺ is seen for He droplets doped with SF₆. The fragmentation patterns for both molecules embedded in He are compared with gas phase studies. Ionization via electron transfer to He⁺ forms highly excited ions that cannot be stabilized by the surrounding He droplet. Besides the atomic fragments F⁺ and Cl⁺ several molecular fragment cations are observed with He atoms attached.     

The Role of Nebulizer Gas Flow in Electrosonic Spray Ionization (ESSI)

In this work, we investigated the role of the nebulizer gas flow in electrosonic spray ionization (ESSI), by systematically studying the relation between the flow and the ion signals of proteins, such as cytochrome c and holomyoglobin using ESSI-mass spectrometry (MS). When a neutral solution was delivered with a small sample flow rate (≤5 μL/min), no obvious transition from electrospray ionization (ESI) to ESSI was found as the gas velocity varies from subsonic to supersonic speed. Droplets mostly experienced acceleration instead of breakup by the high-speed nebulizer gas. On the contrary, using particular experimental conditions, such as an acidic solution or high sample flow rate (≥200 μL/min), more folded protein ions appear to be kept in droplets of diminishing size due to breakup by the high-speed nebulizer gas in ESSI compared with ESI. Theoretical analyses and numerical simulations were also performed to explain the observed phenomena. These systematic studies clarify the ionization mechanism of ESSI and provide valuable insight for optimizing ESSI and other popular pneumatically assisted electrospray ionization methods for future applications.     

Ion tree‐based structure elucidation of acetophenone dimers (AtA) from Acronychia pedunculata and their identification in extracts by liquid chromatography electrospray ionization LTQ‐Orbitrap mass spectrometry

Acronychia‐type acetophenones (AtA) is a chemical group of compounds of important structural and biological interest, abundant in Acronychia species. However, there are no data available for their characterization using mass spectrometry. In the current work, AtA have been investigated by multistage high resolution mass spectrometry and both electrospray ionization and atmospheric pressure chemical ionization, in positive and negative mode, were utilized for their structure elucidation and identification. The analysis of AtA using a linear ion trap‐Orbitrap analyzer enabled the structural determination of key fragment ions and cleavages, which can be used for the structural characterization thereof. A systematic nomenclature based on protonated and deprotonated fragment ions under collision‐induced dissociation conditions and decision trees for the structural determination of AtA are proposed. Furthermore, taking advantage of the characteristic fragmentation patterns, a selective Ultra High Performance Liquid Chromatography Electrospray Ionization multistage Mass Spectrometry (UHPLC‐ESI(‐)‐MSⁿ) method was developed and successfully applied for the dereplication of known AtA and the identification of potentially new ones in Acronychia extracts. Despite the structure similarity and the presence of isomers, accurate characterization of known and unknown AtA derivatives was possible. Copyright © 2015 John Wiley & Sons, Ltd.     

电喷雾萃取电离质谱在线监测 Friedl?nder 缩合反应

质谱在线监测有机化学反应能够实时捕捉反应过程中反应物、中间体、产物的信号及其变化情况,从而推测有机反应机理,并加深对反应本质的认识。本研究利用电喷雾萃取电离质谱(EESI-MS)技术在线监测草酸催化的邻氨基苯乙酮与乙酰丙酮间的Friedlnder缩合反应。在反应的不同时间点,分别捕捉到了反应物(m/z 136)、中间体(m/z 218、m/z 236)和产物(m/z 200)的准分子离子信号,并记录了它们的信号强度随反应时间的变化趋势,推测出了反应机理。     

Monte carlo simulation of macromolecular ionization by nanoelectrospray

Electrospray ionization (ESI) is commonly used in macromolecular mass spectrometry, yet the dynamics of macromolecules in ESI droplets are not well understood. In this study, a Monte Carlo based model was developed, which can predict the efficiency of electrospray ionization for macromolecules, i.e., the number of macromolecular ions produced per macromolecules electrosprayed. The model takes into account ESI droplet evaporation, macromolecular diffusion within the droplet, droplet fissions, and the statistical nature of the ESI process. Two idealized representations of macromolecular analytes were developed, describing cluster prone, droplet surface inactive macromolecules and droplet surface active macromolecules, respectively. It was found that surface active macromolecules are preferentially ionized over surface inactive cluster prone macromolecules when the initial droplet size is large and the analyte concentration in solution is high. Simulations showed that ESI efficiency decreases with increasing initial droplet size and analyte molecular weight, and is influenced by analyte surface activity, the properties of the solvent, and the variance of the droplet size distribution. Model predictions are qualitatively supported by experimental measurements of macromolecular electrospray ionization made previously. Overall, this study demonstrates the potential capabilities of Monte Carlo based ESI models. Future developments in such models will allow for more accurate predictions of macromolecular ESI intensity.     

Lycorine Alkaloid and Crinum americanum L. (Amaryllidaceae) Extracts Display Antifungal Activity on Clinically Relevant Candida Species

Candida species are the main fungal agents causing infectious conditions in hospital patients. The development of new drugs with antifungal potential, increased efficacy, and reduced toxicity is essential to face the challenge of fungal resistance to standard treatments. The aim of this study is to evaluate the in vitro antifungal effects of two crude extracts of Crinum americanum L., a rich alkaloid fraction and lycorine alkaloid, on the Candida species. As such, we used a disk diffusion susceptibility test, determined the minimum inhibitory concentration (MIC), and characterized the components of the extracts using Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (ESI FT-ICR MS). The extracts were found to have antifungal activity against various Candida species. The chemical characterization of the extracts indicated the presence of alkaloids such as lycorine and crinine. The Amaryllidaceae family has a promising antifungal potential. Furthermore, it was found that the alkaloid lycorine directly contributes to the effects that were observed for the extracts and fraction of C. americanum.     

Electrospray ionization mass spectrometry: a major tool to investigate reaction mechanisms in both solution and the gas phase

Electrospray ionization mass spectrometry (ESI-MS), in conjunction with its tandem version ESI-MS/MS, is now established as a major tool to study reaction mechanisms in solution. This suitability results mainly from the ability of ESI to "fish" ions directly from solution to the gas phase environment of mass spectrometers. In this review, we summarize recent studies from our laboratory on the use of on-line monitoring by ESI-MS ion fishing of several types of reactions that permitted us to follow how these reactions progress as a function of both time and conditions using the ultra-high sensitivity and speed of ESI-MS to detect and even characterize transient reaction intermediates. We also show that the intrinsic reactivity of each key gaseous species fished by ESI can be further investigated via ESI-tandem mass spectrometry experiments, searching for the most active species via gas-phase ion/molecule reactions. In the gas-phase, solvent and counter-ion effects are absent. These studies often permit a detailed overview of major steps via the interception, characterization and reactivity investigation of key reaction players.     

The Evolution of Electrospray Generated Droplets is Not Affected by Ionization Mode

Ionization efficiency and mechanism in ESI is strongly affected by the properties of mobile phase. The use of mobile-phase properties to accurately describe droplets in ESI source is convenient but may be inadequate as the composition of the droplets is changing in the plume due to electrochemical reactions occurring in the needle tip as well as continuous drying and fission of droplets. Presently, there is paucity of research on the effect of the polarity of the ESI mode on mobile phase composition in the droplets. In this paper, the change in the organic solvent content, pH, and droplet size are studied in the ESI plume in both ESI+ and ESI– ionization mode. We introduce a rigorous way – the absolute pH (pH_abs ^H ₂ ^O) – to describe pH change in the plume that takes into account organic solvent content in the mobile phase. pH_abs ^H ₂ ^O enables comparing acidities of ESI droplets with different organic solvent contents. The results are surprisingly similar for both ionization modes, indicating that the dynamics of the change of mobile-phase properties is independent from the ESI mode used. This allows us to conclude that the evolution of ESI droplets first of all proceeds via the evaporation of the organic modifier and to a lesser extent via fission of smaller droplets from parent droplets. Secondly, our study shows that qualitative findings related to the ESI process obtained on the ESI+ mode can almost directly be applied also in the ESI– mode.

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Determination of linear alkylbenzenesulfonates in communal wastewater by means of solid phase microextraction coupled with API-MS and HPLC-FLD

The use of Solid Phase Microextraction (SPME) for the qualitative and quantitative determination of Linear Alkylbenzenesulfonates (LAS) in waste water samples was investigated. A Carbowax/Templated Resin (CW/?TPR) coated fiber was directly immersed into influent and effluent samples of a sewage treatment plant (STP). Extraction conditions such as time, pH, ion strength were investigated. The extracted LAS were desorbed with a solvent in a specially designed SPME-LC interface for analysis with HPLC-FLD and Electrospray Ionization Mass Spectrometry (ESI-MS). The combination of SPME with ESI-MS proved to be an alternative technique for the LAS determination in waste water. Linear ranges of the external calibration were found from 0.5–100 ng/mL, with detection limits of 0.5 ng/mL for each individual LAS homologue. The reproducibility of the method is 15% (relative standard deviation). Received: 3 May 1999 / Revised: 22 July 1999 / Accepted: 24 July 1999