Electrosonic Spray Research Articles

Desorption Electrospray Ionization Mass Spectrometry: 20 Years.

ConspectusMass spectrometry (MS) is one of the most widely used technologies in the chemical sciences. With applications spanning the monitoring of reaction products, the identification of disease biomarkers, and the measurement of thermodynamic parameters and aspects of structural biology, MS is well established as a universal analytical tool applicable to small compounds as well as large molecular complexes. Regardless of the application, the generation of gas-phase ions from neutral compounds is a key step in any MS experiment. However, this ionization step was for many years limited to high-energy approaches that required gas-phase analytes and thus it was restricted to volatile samples. Over the last few decades, new methodologies have been developed to address this limitation and facilitate ionization of biological molecules. Electrospray ionization (ESI) is the most broadly used of these methods, as it facilitates the ionization of intact polar compounds from solution.Twenty years ago, our group reported a new ionization method that uses a charged solvent spray to impact a surface, generating ions from objects rather than just solutions and doing so directly in the ambient environment with no vacuum requirements and little to no sample preparation. This method was termed desorption electrospray ionization (DESI), and it initiated a new field that would come to be known as ambient mass spectrometry. The simplicity and wide applicability of the DESI technology─and the tens of ambient ionization methods developed subsequently─revolutionized the MS analysis of complex materials for their organic components, especially for in situ applications.This Account describes the history of DESI, starting with the development of the technique from early electrosonic spray ionization (ESSI) experimental observations as well as the studies leading to the understanding of its mechanism as a "droplet pick-up" phenomenon involving sequential events (i.e., thin film formation, solid-liquid extraction, secondary droplet generation, and ESI-like ionization from these droplets). We also overview the developments and applications of the technology that have been demonstrated by our group during the last two decades. In particular, we describe (i) the use of DESI for tissue imaging, one of its more significant applications to date, and its extension to intraoperative clinical diagnosis; (ii) the integration of the technology with portable instrumentation for in situ analysis, especially when coupled with tandem mass spectrometry (MS/MS); (iii) the use of DESI microdroplets as microvessels to accelerate organic reactions by orders of magnitude compared to those in bulk solution; and (iv) the combination of all these capabilities for automated high-throughput experiments aimed at accelerating drug discovery.

Real-time bottom-up characterization of protein mixtures enabled by online microdroplet-assisted enzymatic digestion (MAED).

Enzymatic digestion remains one of the "rate-determining steps" in the bottom-up analysis of proteins. However, by performing digestion in microdroplets generated from electrosonic spray, the reaction could be accelerated to a timescale lower than milliseconds. Here, we describe a simple and rapid online digestion platform named online microdroplet-assisted enzymatic digestion (MAED). It involves the integration of intact protein separation with enzymatic digestion in microdroplets. Via online MAED, various protein standards, including an antibody standard, were characterized in a bottom-up manner without prior digestion, and high sequence coverages were obtained. We further extended the application of online MAED to a more complex sample, mouse brain extract, where protein identifications were successfully yielded. Compared with the conventional bottom-up approach, a more comprehensive characterization could be obtained particularly for low molecular weight proteins. In short, we provide a rapid and alternative bottom-up analysis in a top-down fashion as well as a new possibility for microdroplet chemistry.

OH radical-initiated single-electron transfer for accelerated degradation via carbocation intermediates.

Single electron transfer (SET) has made great contributions to a broad range of chemical processes, whose radical cation and carbocation intermediates are important for mechanism studies. Herein, hydroxyl radical (˙OH)-initiated SET was revealed in accelerated degradations, via the online examination of radical cations and carbocations by electrosonic spray ionization mass spectrometry (ESSI-MS). In the green and efficient non-thermal plasma catalysis system (MnO2-plasma), hydroxychloroquine was efficiently degraded upon SET via carbocations. In the plasma field full of active oxygen species, ˙OH was generated on the MnO2 surface to initiate SET-based degradations. Furthermore, theoretical calculations revealed that ˙OH preferred to withdraw the electron from the N atom that was conjugated to the benzene ring. This facilitated the generation of radical cations through SET, which was followed by the sequential formation of two carbocations for accelerated degradations. Transition states and energy barriers were calculated to study the formation of radical cations and subsequent carbocation intermediates. This work demonstrates an ˙OH-initiated SET for accelerated degradation via carbocations, providing a deeper understanding and the potential for the wider application of SET in green degradations.

Helium assisted desorption and spray ionization

We introduce a new method, helium assisted desorption ionization, that uses helium as nebulizing gas in desorption electrospray ionization (DESI). We also investigate Helium Assisted Spray Ionization, when helium is used with a micro-electrospray ionization source similar to electrosonic spray ionization (ESSI). In both methods, analyte ion signals are improved, while the undesirable effects of species created by the breakdown products of atmospheric gases and helium are suppressed. Restriction provided by the extended ion inlet capillary reduces the influx of helium, preventing a deleterious increase in operating pressure in the intermediate ion transfer region. We demonstrate significant improvements in signal intensities for analytes of different sizes and polarities, including xanthine alkaloids, lipids, and proteins. Compared to regular DESI, ion intensities for both protein and small molecules increased with helium addition, while less-polar small molecules were improved to a greater extent. The improvements in signal response with helium nebulization generally followed the same trend using either helium assisted desorption or spray ionization, although the extent of improvement was more substantial in the desorption experiments. This observation suggests that desorption, ionization, and/or ion transport processes could be positively influenced when helium is used as nebulizing gas. Nebulization with helium also produces a smaller desorption footprint compared to nitrogen, promising benefits for chemical microscopy and other applications of DESI-MS.

Microdroplet Mass Spectrometry Enables Extremely Accelerated Pepsin Digestion of Proteins.

In microdroplets, rates of chemical or biomolecular reactions can exceed those in the bulk phase by more than a million times. As electrospray ionization-based mass spectrometry (MS) involves the formation of charged microdroplets, reaction acceleration and online MS monitoring of reaction products can readily be performed at the same time. We investigated accelerated enzymatic reactions in microdroplets and focused on the proteolytic enzyme pepsin. Electrosonic spray ionization (ESSI) was utilized for developing the ultrarapid pepsin in-spray digestion of two different proteins, cytochrome c and RocC, at low pH values. The optimization of the protein digestion aimed at achieving maximum sequence coverage for the analyzed proteins. Furthermore, carefully designed control experiments allowed us to unambiguously prove that enzymatic protein cleavage almost exclusively occurs within the spray at a millisecond time scale and not prior to microdroplet generation.

Accelerated Oxidation of Organic Sulfides by Microdroplet Chemistry.

We report the rapid oxidation of organic sulfides to sulfoxides by means of microdroplet chemistry at room temperature using a spray solution containing an organic sulfide dissolved in water/methanol, dilute (11%-14%) sodium hypochlorite (NaClO), and 5% chloroauric acid (HAuCl4). Ultrasonic nebulization, easy ambient sonic-spray ionization, or electrosonic spray ionization serves as the microdroplet source. High-resolution mass spectrometry was used as an online detector, and nuclear magnetic resonance was used as an offline detector. We found that the sulfoxide yields vary between 66 and 95%, the highest rate of product formation is 195 mg/min for benzyl phenyl sulfoxide, and the time required is a few minutes, which is much less than that required for the conventional means of achieving this chemical transformation. We also applied this microdroplet method to protein fingerprinting. We found that protein sequences containing methionine can be quickly oxidized, providing useful information for protein structure determinations.

In situ probing changes in fatty-acyl chain length and desaturation of lipids in cancerous areas using mass spectrometry imaging.

Aberrant changes in the expression levels and structure of lipids may shape tumor microenvironment. In this study, we have performed mass spectrometry imaging and profiling analysis of 63 tissues of five types of cancer, namely, breast, colorectal, esophageal, lung, and gastric cancer, using in situ liquid extraction electrosonic spray ionization mass spectrometry imaging. Alteration of fatty-acyl chain length of unsaturated phosphatidylcholines, phosphatidylinositols, and phosphatidylserines and of chain length of (un)saturated fatty acids are associated with different cancerous areas of five types of cancer. The ratios of the same fatty-acyl carbon atom lipids with one double bond difference and the ratios of the same chain-length fatty acids with one double bond difference exhibited significant differences among the cancerous areas of five types of cancer. Our data may reveal that there were different lipid metabolism networks among five types of cancer.

A critical analysis of electrospray techniques for the determination of accelerated rates and mechanisms of chemical reactions in droplets.

Electrospray and Electrosonic Spray Ionization Mass Spectrometry (ESI-MS and ESSI-MS) have been widely used to report evidence that many chemical reactions in micro- and nano-droplets are dramatically accelerated by factors of ∼102 to 106 relative to macroscale bulk solutions. Despite electrospray's relative simplicity to both generate and detect reaction products in charged droplets using mass spectrometry, substantial complexity exists in how the electrospray process itself impacts the interpretation of the mechanism of these observed accelerated rates. ESI and ESSI are both coupled multi-phase processes, in which analytes in small charged droplets are transferred and detected as gas-phase ions with a mass spectrometer. As such, quantitative examination is needed to evaluate the impact of multiple experimental factors on the magnitude and mechanisms of reaction acceleration. These include: (1) evaporative concentration of reactants as a function of droplet size and initial concentration, (2) competition from gas-phase chemistry and reactions on experimental surfaces, (3) differences in ionization efficiency and ion transmission and (4) droplet charge. We examine (1–4) using numerical models, new ESI/ESSI-MS experimental data, and prior literature to assess the limitations of these approaches and the experimental best practices required to robustly interpret acceleration factors in micro- and nano-droplets produced by ESI and ESSI.

Accelerated tert-butyloxycarbonyl deprotection of amines in microdroplets produced by a pneumatic spray

Protection and deprotection of organic compounds in multistep reactions using functional groups such as tert-butoxycarbonyl (Boc), is widely performed in synthetic organic chemistry. Reaction rate acceleration studies in spray-based ionization methods (electrosonic spray, paper spray, nanospray) have become increasingly common. Here, we demonstrate reaction rate acceleration of Boc deprotection using easy ambient sonic-spray ionization (EASI), a pneumatic technique which does not involve an applied voltage, in a teaching laboratory setting. The goal of this laboratory exercise was to explore acceleration in a previously unexplored spray-based reaction, while emphasizing in a pedagogic setting the importance of protecting groups for multistep synthesis. Rate acceleration factors of more than an order of magnitude were observed in the uncharged micron-sized droplets generated by EASI. The effect of reaction conditions on reaction acceleration was examined including changes in the type of acid, reagent concentration ratios and syringe pump flow rates. Student knowledge was assessed by pre-laboratory assignments, post-laboratory reports and oral interviews.

Monitoring changes of docosahexaenoic acid-containing lipids during the recovery process of traumatic brain injury in rat using mass spectrometry imaging

Brain lipid homoeostasis is critical during recovery process after traumatic brain injury (TBI). In this study, we integrated liquid extraction and electrosonic spray ionization technology to develop an ionization device coupled with a Fourier transform ion cyclotron resonance mass spectrometer for imaging of docosahexaenoic acid (DHA)-containing lipids on rat brain tissues. The ion images of the brain tissue sections from the normal rats and the rats after TBI at acute phase (0 and 1 day) and chronic phase (3, 5, and 7 days) were obtained. The imaging results indicate that the levels of DHA and lyso-phosphatidylethanolamine (22:6) in the injury area of TBI rats increased significantly at the acute phase and subsequently decreased at the chronic phase. But the levels of DHA-containing phospholipids including phosphatidylethanolamine (PE)(P-18:0/22:6), PE(18:0/22:6), and phosphatidylserine (18:0/22:6) decreased at the acute phase and gradually increased at the chronic phase in the injury area accompanied by the morphogenesis and wound healing. These findings indicate that the DHA may participate in the recovery process of brain injury. This is the first report to in situ detect the changes in the levels of DHA and DHA-containing lipids in the TBI model.

Role of Electrochemistry in Desorption Ionization Mass Spectrometry (LS DESI MS) of Aqueous Samples Containing Electrolyte Salts.

The ionization of LS samples in desorption ionization mass spectrometry (LS DESI MS), supplied continuously through a LS interface separated in space from the spray emitter, was investigated in this work. The role of electrochemistry (EC) in the ionization process was addressed. The visual (observation) of the operation of the LS DESI MS system showed a thick spray plume generated by the electrosonic spray ionization (ESSI), forming a liquid cone at the LS interface. When the LS interface was grounded the cone collapsed and the MS ion signal was lost, indicating that the LS was carried to the MS inlet by the spray that emerged from the cone. Ion signals in a new in-line LS DESI MS system, in angled LS DESI MS, and in electrospray ionization (ESI) MS, which produced the most intense ion signals from methanol/water solutions, and in ESSI MS, of dopamine (DA), tyrosine (Tyr) and N,N-dimethyl-p-phenylenediamine (DMPA), were evaluated using methanol/water and aqueous (aq) solutions. In addition, the effect on ion signals of geometric parameters and the LS and the spray solution flow rates was tested in in-line LS DESI MS. Of the methods tested, the analysis of aq LS containing electrolytes was simplest by LS DESI MS. The signal intensity was higher in in-line than in angled LS DESI MS. In online electrochemistry (EC)/LS DESI MS, when 0 V was applied to the EC cell Tyr ion signal was detected only at low pH (2).

Accelerated hydrazone formation in charged microdroplets.

Electrospray ionization-mass spectrometry (ESI-MS) is an emerging tool for reaction monitoring. It can be accompanied by reaction acceleration in charged droplets. The time course of the bulk reaction of indoline-2,3-dione with phenylhydrazine in methanol to produce 3-(2- phenylhydrazono)indolin-2-one was monitored by ESI. Both nanoESI and electrosonic spray ionization (ESSI) were used for this study as representing two common forms of ionization for reaction monitoring. The effect on product yield of the distance the droplets travel between the source and the MS inlet was varied and product/starting material ratios were examined. Product yield is dramatically increased by increasing the distance. At short distances reaction monitoring can be performed without acceleration and at greater distances reaction acceleration occurs. This distance effect over the course of the reaction roughly parallels the time dependence of the bulk-phase reaction. Reaction acceleration in droplets is attributed to solvent evaporation leading to increased surface to volume ratios. An acceleration factor of 10(4) , measured relative to the bulk reaction at short times, is readily achieved by simply increasing the droplet distance of flight showing that the same ionization source can be used to monitor reactions with or without acceleration. Copyright © 2016 John Wiley & Sons, Ltd.

Depth-Resolved Nanospray Desorption Electrospray Ionization Mass Spectroscopy in Electrochemically Active Biofilms

Recently, some bacterial biofilms have been shown to colonize and transfer their extracellular electrons to solid electrodes in microbial fuel cells and in bioelectrochemical systems [1, 2]. This class of microorganisms is described as electrochemically active biofilms (EABs) that are able to transfer electrons outside the cell to insoluble electron acceptors (iron or metal oxides) or to solid electrodes via extracellular electron transfer [3, 4]. The need to understand the distribution of proteins involved in electron transfer in EABs is of critical importance. These proteins can be identified using mass spectroscopy (MS). It is known that there is a wide spectrum of applications in which mass spectroscopy (MS) can provide information from a sample of EABs, and these applications can easily translate to all areas of research in EABs. However, the spatial diversity of microbial functions within EABs cannot be resolved because all of the current MS analysis techniques are conducted in the bulk liquid, which provides no spatial resolution with respect to the structure of EABs. Separation techniques can be combined with MS to present a more focused data set with regard to composition; however, MS analysis still lacks the capacity to investigate variation with depth of EABs. Depth profiles will have the potential to elucidate the mechanisms of the surrounding matrix and the roles of the microorganism with respect to the layers of EABs and the growth interface. To date, this MS technique has not been used for depth profiling in EABs. This is mostly because there was no tool available to extract samples from different depths in EABs. Microcapillaries with a several-micrometer tip diameter can be used in EABs without damaging its structure for the depth profiling of selected chemicals. The goal of this work was to develop a microcapillary system which can extract samples at desired depth inside the EABs. Ambient pressure surface ionization mass spectrometry is used to obtain a chemical analyte for sampling from interfaces without special sample preparation [5]. Desorption electrospray ionization (DESI) is an ambient ionization technique in which charged droplets from an electrosonic spray ionization source are aimed towards a surface with a proximal atmospheric pressure mass spectrometer inlet. In this technique, analyte molecules are collected from flat surfaces followed by ionization using a self-aspirating nanoelectrospray. This technique directly transports and ionizes an analyte that is desorbed from a surface into a liquid and it is called as nanospray DESI (nano-DESI). The nanospray capillary transports the charged liquid to the mass spectrometer inlet directly, eliminating splashing while minimizing analyte transport distance. The target application of the developed microcapillary system is to interface it with a nano-DESI sensor which can be used to characterize in situ, depth-resolved analyses of metabolites and possibly proteins. The developed nano-DESI sensor is composed of two microcapillaries placed in an outer case made of glass. Figure 1 shows an image of a developed nano-DESI sensor. In this system, the solvent delivery and its collection were made from the same microcapillaries. We managed to operate the sensor between 10 nL/min and 100 mL/min flow rates. After optimization of the flow rates, we tested it in EABs. EABs were grown according to our previously published paper and book [6, 7]. While we had succeeded in developing a nano-DESI sensor, we found unexpected challenges using it with EABs. Finally, the microcapillary system developed for this work enabled us to use it for quantifying electron transfer processes in EABs [7].

Alleviation of ion suppression effect in sonic spray ionization with induced alternating current voltage.

In this study, alleviation of ion suppression effect in sonic spray ionization mass spectrometry (SSI-MS) was investigated. Ion suppression effect was firstly compared between electrospray ionization (ESI) and conventional SSI, and more severe ion suppression effect was observed with SSI. Ion suppression effect of SSI was also found difficult to be alleviated by simply optimizing major parameters. Alternatively, we found that with the assistance of an alternating current (AC) voltage with low amplitude, the ion suppression effect was greatly alleviated (comparable with conventional ESI). That AC voltage was applied outside the SSI spray tip, and no direct contact between the electrode and spray solution was necessary. Besides the alleviation of the ion suppression effect, this newly-developed method, termed as induced electrosonic spray ionization (IESSI), appeared to preserve similar charge state distribution with SSI for protonated cytochrome c, hemoglobin, and bradykinin. IESSI could also obtain significantly improved ion intensities (~1000-fold over conventional SSI). In addition, tolerance of concentrated salts for IESSI-MS was investigated through the analysis of cytochrome c in the presence of concentrated sodium chloride (NaCl) or ammonium acetate (NH4 OAc).

Modes of Activation of Organometallic Iridium Complexes for Catalytic Water and C–H Oxidation

Sodium periodate (NaIO4) is added to Cp*Ir(III) (Cp* = C5Me5(-)) or (cod)Ir(I) (cod = cyclooctadiene) complexes, which are water and C-H oxidation catalyst precursors, and the resulting aqueous reaction is investigated from milliseconds to seconds using desorption electrospray ionization, electrosonic spray ionization, and cryogenic ion vibrational predissociation spectroscopy. Extensive oxidation of the Cp* ligand is observed, likely beginning with electrophilic C-H hydroxylation of a Cp* methyl group followed by nonselective pathways of further oxidative degradation. Evidence is presented that the supporting chelate ligand in Cp*Ir(chelate) precursors influences the course of oxidation and is neither eliminated from the coordination sphere nor oxidatively transformed. Isomeric products of initial Cp* oxidation are identified and structurally characterized by vibrational spectroscopy in conjunction with density functional theory (DFT) modeling. Less extensive but more rapid oxidation of the cod ligand is also observed in the (cod)Ir(I) complexes. The observations are consistent with the proposed role of Cp* and cod as sacrificial placeholder ligands that are oxidatively removed from the precursor complexes under catalytic conditions.

Venturi-Electrosonic Spray Ionization Cataluminescence Sensor Array for Saccharides Detection

In this article, a Venturi electrosonic spray ionization (V-ESSI) cataluminescence (CTL) sensor array was reported for discriminating saccharides in solution. Integrating electrosonic spray ionization (ESSI), a liquid system of Venturi self-pumping injection for the CTL reaction, was fabricated for enhancing CTL reactivity of aqueous samples. Comparing with simple Venturi injection by air and Venturi easy ambient sonic-spray ionization without electric assistance (V-EASI), the remarkable enhancement of CTL signals resulted from V-ESSI. This system showed higher cross-reactive CTL responses catalyzed by alkaline earth metal-nanomaterials than other catalysts, giving different signals for a given saccharide on different catalysts and different responses for different saccharides on the same catalyst. Then, a 4 × 2 CTL sensor array was used for obtaining "fingerprints" of distinct CTL response patterns. Analyzed by linear discriminant analysis (LDA), this V-ESSI CTL sensor array not only achieved the well discrimination of different saccharides (99.9% of total variation) but also discriminated four groups of urine sugar-level for urine samples from diabetic patients (98.1% of discrimination accuracy). It had good reproducibility and gave a linear range of 22.5-67558 μg/mL (R > 0.99) for xylose with a detection limit of 7.4 μg/mL on MgO. As a new artificial tongue, this system provided a simple, rapid, low cost, low energy consumption, and environmentally friendly pathway for aqueous sample discrimination. It has dramatically expanded applications of the CTL-based senor array and will be applicable to clinical diagnoses, environment monitoring, industrial controls, food industry, and various marine monitoring.

Signal and charge enhancement for protein analysis by liquid chromatography–mass spectrometry with desorption electrospray ionization

We recently reported the use of desorption electrospray ionization (DESI) as a novel interface to couple high-performance liquid chromatography (HPLC) with mass spectrometry (MS) (Chem. Commun. 47 (2011) 4171). One of the benefits of such an interface is that post-column derivatization of separated analytes can be integrated with ionization via a “reactive” DESI approach in which a derivatizing reagent is doped into the spray solvent. The reactive DESI interface allows analyte desorption/ionization from the end of the chromatographic column with prompt MS detection; a short time delay of ∼20ms was demonstrated. In this study, we extended this application by “supercharging” proteins following HPLC separation using a DESI spray solvent containing supercharging reagents, m-nitrobenzyl alcohol (m-NBA) or sulfolane. Proteins (insulin, ubiquitin, lysozyme and α-lactalbumin) eluted out of the LC column can be supercharged with the protein charge state distributions (CSDs) significantly increased (to higher charge), which would be advantageous for subsequent top-down MS analysis of proteins. Interestingly, supercharging combined with reactive DESI enhances tolerance towards trifluoroacetic acid (TFA), which is known to be a superior additive in the mobile phase for premium peptide/protein chromatographic separation but has severe signal suppression effects for conventional electrospray ionization (ESI). In comparison to electrosonic spray ionization (ESSI), a variant form of ESI, the sensitivity of protein analysis using LC/DESI-MS with the mobile phase containing TFA can be improved by up to 70-fold for lysozyme and α-lactalbumin by including m-NBA in the DESI spray solvent. Presumably, by reducing TFA dissociation in the droplet, supercharging agents lower trifluoroacetate anion concentrations and concomitantly reduce ion pairing to analyte cationic sites. The reduced ion pairing therefore decreases the TFA signal suppression effect. The supercharging capability and the reduction of TFA signal suppression suggest that LC/DESI-MS is a valuable method for protein analysis.

Gas-phase basicity of several common MALDI matrices measured by a simple experimental approach

We present a combined experimental and theoretical study to determine the gas-phase basicity of several relatively small organic molecules that are used as matrices in MALDI: alpha-cyano-4-hydroxycynnamic acid (αCHCA), ferulic acid (FA), 3-hydroxypicolinic acid (HPA), sinapinic acid (SA), and 2,4,6-trihydroxyacetophenone (THAP). Our simple and fast experimental approach utilizes the ability of an electrosonic spray ionization (ESSI) source to produce high densities of gas-phase ions at ambient conditions, so that their gas-phase thermochemical properties can be probed in a bracketing-like experiment at atmospheric pressure, with no need for high vacuum. The reaction between the analyte and reference bases is monitored by a commercial mass spectrometer. We determined the following GB values of the MALDI matrices studied: 908.1 ± 0.8 (αCHCA), 863.8 ± 8.0 (FA), 876.3 ± 10.2 (HPA), 867.4 ± 1.4 (SA), and 864.2 ± 7.6 (THAP) kJ mol−1. A good agreement between the experimentally measured and computationally predicted (DFT-B3LYP, G4MP2) gas-phase basicities for the studied compounds is found, except for αCHCA for which the calculations resulted in approximately 80 kJ mol−1 lower GB. We attribute the observed discrepancy between the measured and calculated values to a gas-phase rearrangement of αCHCA into a heterocyclic aromatic isomer, 7-hydroxyquinoline-3-carboxylic acid (HQA). HQA is thermodynamically more stable and features higher GB than that of αCHCA (calculated at approximately 920 kJ mol−1). The gas-phase basicity values of THAP and HPA are reported for the first time.

Studies of Heterogeneous/Homogeneous Ion-Molecule Reactions by Ambient Ionization Mass Spectrometry

The general concepts, features and applications of ambient ionization mass spectrometry for studying the heterogeneous/homogeneous ion-molecule reactions are reviewed. Important ambient ionization mass spectrometric methods and their applications in organic chemistry are summarized, focusing on SESI (secondary electrospray ionization), DESI (desorption electrospray ionization-mass spectrometry), ESSI (electrosonic spray ionization), EESI (extractive electrospray ionization-mass spectrometry), ELDI (electrospray-assisted laser desorption/ionization), LTP (low-temperature plasma probe) and APTDI (atmospheric-pressure thermal desorption ionization) technologies. At the same time, the advantages of ambient ionization mass spectrometry in studying organic reactions allow chemists to explore new type of organic reactions, probe reaction mechanisms and to intercept/characterize transient reactive intermediates in ambient condition. Keywords: Ambient mass spectrometry, ionization, desorption, transient reactive intermediates, heterogeneous/homogeneous ion-molecule reaction, Gas phase ion-molecule reaction, analytical technique, organic reactions, atom bombardment, atmospheric pressure, intrinsic, Electrospray Ionization, Nebulizing Gas, unsaturated lipid, ozone

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 (≤5μL/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 (≥200μL/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.