Ambient Ionization Methods Research Articles

Identification of Plant Compounds with Mass Spectrometry Imaging (MSI).

The presence and localization of plant metabolites are indicative of physiological processes, e.g., under biotic and abiotic stress conditions. Further, the chemical composition of plant parts is related to their quality as food or for medicinal applications. Mass spectrometry imaging (MSI) has become a popular analytical technique for exploring and visualizing the spatial distribution of plant molecules within a tissue. This review provides a summary of mass spectrometry methods used for mapping and identifying metabolites in plant tissues. We present the benefits and the disadvantages of both vacuum and ambient ionization methods, considering direct and indirect approaches. Finally, we discuss the current limitations in annotating and identifying molecules and perspectives for future investigations.

Criticality of Spray Solvent Choice on the Performance of Next Generation, Spray-Based Ambient Mass Spectrometric Ionization Sources: A Case Study Based on Synthetic Cannabinoid Forensic Evidence

Mass spectrometry (MS) is a highly selective and sensitive analytical tool with a myriad of applications, but such techniques are typically used in laboratory settings due to the handling and preparations that are necessary. The merging of two streams of robust research, portable MS systems and next-generation ambient ionization methods, now provides the ability to perform high-performance chemical screening in an on-site and on-demand manner, with natural applications in disciplines such as forensic science, where samples of interest are typically found in field environments (i.e., traffic stops, crime scenes, etc.). Correspondingly, investigations regarding the suitability and robustness of these methodologies when they are utilized for authentic forensic evidence processing are prudent. This work reports critical insights into the role that choice of spray solvent system plays regarding analytical performance of two spray-based ambient ionization sources, paper spray ionization (PSI) and filter cone spray ionization (FCSI), when employed for evidence types containing emerging synthetic cannabinoids. The systematic characterization studies reported herein show that the applied spray solvent can dramatically affect both spectral intensity and signal duration, and in some circumstances, yield deleterious false negative responses. Overall, acetonitrile-based systems are shown to strike a balance between analyte solubility concerns and spray ionization dynamics of the novel ion sources employed on portable MS systems.

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.

Exploring new dimensions: Single and multi-block analysis of essential oils using DBDI-MS and FT-IR for enhanced authenticity control

BackgroundEssential oils (EOs) are complex mixtures of volatile hydrocarbons with a wide range of applications in the pharmaceutical, fragrance and food industry. The composition of EOs is highly variable and can affect their quality and pharmaceutical efficacy. Moreover, the high economic value of EOs, such as those obtained from Rosa damascena, make falsification and misclassification a lucrative business. Consequently, adulterations can lead to serious health consequences for consumers. While current quality control methods for EOs involve analysing their chromatographic profile or comparing their Fourier transform infrared (FT-IR) spectra, these methods can be time-consuming or lack sensitivity. To address these issues, we compared state-of-the-art quality control methods, including gas chromatography flame ionization detection (GC-FID) quantification and enantiomeric ratio determination, FT-IR spectrometry with dielectric barrier discharge ionization coupled to triple quadrupole mass spectrometer (DBDI-MS), in a chemometric single- and multi-block approach. ResultsOur results show that the best classification accuracy of 94.7% for R. damascena samples was obtained using GC-FID combined with partial least square discriminant analysis (PLS-DA). Comparatively, the enantiomeric ratios did not improve classification accuracy. In contrast, fragmentation data from DBDI-MS (Q3), which was acquired in a fraction of the analysis time and without extensive sample preparation, achieved a classification accuracy of 84.2%. We also found that combining FT-IR with parent ion DBDI-MS (Q1) data in a multi-block sequentially orthogonalized partial least squares linear discriminant analysis (SO-PLS-LDA) model improved classification accuracy, compared to their respective single-block PLS-DA models. SignificanceOverall, our study demonstrates that DBDI, as an ambient ionization method, has significant potential for high-throughput screening. When combined with MS, it can produce comparable classification accuracies to conventional methods, while offering the added benefits of speed and convenience. As such, DBDI-MS is a promising tool for EO quality control.

LMJSSP for analysis of prophylactic lubricants, spermicides and residues

DNA evidence in sexual assault cases have proven increasingly difficult to obtain and analyse due to increased condom use. With more interest in alternatives to DNA evidence, prophylactic lubricants, spermicides and residues may be interesting prospects. Current interest in the analysis of prophylactic residues focuses on the evaluation and identification of lubricants and constituents, primarily through gas chromatography or Fourier transform infrared spectroscopy. Though cost-effective methods, extensive sample preparation and destructive modes of analysis remain an area for improvement. As a result, the focus has since shifted to ambient ionization methods that offer adequate sensitivity and reduced sample preparation. The Liquid Microjunction Surface Sampling Probe (LMJSSP) is a versatile ambient ionization source that employs a probe that supports a continuously flushing droplet that extracts analytes when placed in contact with a surface. The analytes are aspirated into the mass spectrometer with a Venturi pressure. In this work we use the LMJSSP to analyse the trace transfer of condom lubricant to different types of fabric (cotton, cotton-spandex, and denim). Furthermore, we examine the sensitivity and storage conditions for the direct analysis method on different swab types (cotton, silicone, and foam). Additionally, Principal Component Analysis (PCA) and Maximally Collapsing Metric Learning (MCML) are utilized for visualization of differentiability of commercially available condom brands including Durex™ and Trojan™, and product subtypes. The results present an interesting multi-disciplinary approach of using a direct liquid extraction ambient ionization technique and machine learning to improve the overall workflow for the analysis of lubricants, swabs and fabrics. Machine learning algorithms were able to differentiate between inherent differences of Durex™ and Trojan™ condoms.

New Platform for Label-Free, Proximal Cellular Pharmacodynamic Assays: Identification of Glutaminase Inhibitors Using Infrared Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry.

Cellular pharmacodynamic assays are crucial aspects of lead optimization programs in drug discovery. These assays are sometimes difficult to develop, oftentimes distal from the target and frequently low throughput, which necessitates their incorporation in the drug discovery funnel later than desired. The earlier direct pharmacodynamic modulation of a target can be established, the fewer resources are wasted on compounds that are acting via an off-target mechanism. Mass spectrometry is a versatile tool that is often used for direct, proximal cellular pharmacodynamic assay analysis, but liquid chromatography-mass spectrometry methods are low throughput and are unable to fully support structure-activity relationship efforts in early medicinal chemistry programs. Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is an ambient ionization method amenable to high-throughput cellular assays, capable of diverse analyte detection, ambient and rapid laser sampling processes, and low cross-contamination. Here, we demonstrate the capability of IR-MALDESI for the detection of diverse analytes directly from cells and report the development of a high-throughput, label-free, proximal cellular pharmacodynamic assay using IR-MALDESI for the discovery of glutaminase inhibitors and a biochemical assay for hit confirmation. We demonstrate the throughput with a ∼100,000-compound cellular screen. Hits from the screening were confirmed by retesting in dose-response with mass spectrometry-based cellular and biochemical assays. A similar workflow can be applied to other targets with minimal modifications, which will speed up the discovery of cell active lead series and minimize wasted chemistry resources on off-target mechanisms.

Rapid screening of high-priority N-nitrosamines in pharmaceutical, forensic, and environmental samples with paper spray ionization and filter cone spray ionization-mass spectrometry.

The burgeoning concern of N-nitrosamine (NAM) contamination found in various pharmaceutical compositions has increased the demand for rapid and reliable screening methods to better assess the breadth of the problem. These carcinogenic compounds are also found in food, water, and soil, and they have been used in poison-related homicides. A combination of complementary, ambient ionization methods, paper spray ionization (PSI) and filter cone spray ionization (FCSI)-mass spectrometry (MS), was characterized towards trace-level residue screening of select NAMs (e.g., N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosodibutylamine) directly from complex and problematic matrices of interest, including prescription and over-the-counter tablets, drinking water, soil, and consumable goods. Spectral data for analyte confirmation and detection limit studies were collected using a Thermo LCQ Fleet ion trap mass spectrometer. PSI-MS and FCSI-MS readily produced mass spectral data marked by their simplicity (e.g., predominantly protonated molecular ions observed) and congruence with traditional electrospray ionization mass spectra in under 2min. per sample. Both methods proved robust to the complex matrices tested, yielding ion signatures for target NAMs, as well as active pharmaceutical ingredients for analyzed tablets, flavorants inherent to food products, etc. Low part-per-million detection limits were observed but were shown dependent on sample composition. PSI-MS and FCSI-MS were successful in detecting trace-level NAMS in complex liquid- and solid-phase matrices with little to no prior preparation. This work suggests that these methodologies can provide a means for assessing problematic pharmaceutical adulterants/degradants for expedited quality control, as well as enhancing environmental stewardship efforts and forensic investigations.

Toward Depth-Resolved Analysis of Plant Metabolites by Nanospray Desorption Electrospray Ionization Mass Spectrometry.

Nanospray desorption electrospray ionization (nano-DESI) is one of the ambient desorption ionization methods for mass spectrometry (MS), and it utilizes a steady-state liquid junction formed between two microcapillaries to directly extract analytes from sample surfaces with minimal sample damage. In this study, we employed nano-DESI MS to perform a metabolite fingerprinting analysis directly from a Hypericum leaf surface. Moreover, we investigated whether changes in metabolite fingerprints with time can be related to metabolite distribution according to depth. From a raw Hypericum leaf, the mass spectral fingerprints of key metabolites, including flavonoids and prenylated phloroglucinols, were successfully obtained using ethanol as a nano-DESI solvent, and the changes in their intensities were observed with time via full mass scan experiments. In addition, the differential extraction patterns of the obtained mass spectral fingerprints were clearly visualized over time through selected ion monitoring and pseudo-selected reaction monitoring experiments. To examine the correlation between the time-dependent changes in the metabolite fingerprints and depth-wise metabolite distribution, we performed a nano-DESI MS analysis against leaves whose surface layers were removed multiple times by forming polymeric gum Arabic films on their surfaces, followed by detaching. The preliminary results showed that the changes in the metabolite fingerprints according to the number of peelings showed a similar pattern with those obtained from the raw leaves over time.

An 241Am Plasma Desorption Ionization (AmDI) Source Scavenged from Smoke Detectors for Ambient Mass Spectrometry Sampling.

The development of efficient, low-cost, easy-to-use ambient ionization methods has been a major goal of modern mass spectrometry. In this Letter, we present a gas-free, voltage-free, economic, and safe desorption ionization method using the plasma generated by a radioactive element, americium-241, scavenged from smoke detectors that equip almost every household. No other energy sources, such as laser, discharge, fast-moving carrier gas, solvent droplet, ultrasound, or heat are needed. We name this new method as americium-241 desorption ionization (AmDI). AmDI is tested for the detection of more than 20 volatile and nonvolatile chemicals under different sampling conditions, and the detection limit can be in the range of tens of picograms for some analytes. Mechanistically, we provide evidence that the α particles emitted from radioactive decay ionize ambient air, and the resulting plasma further energizes and ionizes the surface analytes for mass spectrometry detection. We anticipate wide applications of AmDI in mass spectrometric sampling in the near future because of the plethora of merits.

SDHB knockout and succinate accumulation are insufficient for tumorigenesis but dual SDHB/NF1 loss yields SDHx-like pheochromocytomas.

SUMMARYInherited pathogenic succinate dehydrogenase (SDHx) gene mutations cause the hereditary pheochromocytoma and paraganglioma tumor syndrome. Syndromic tumors exhibit elevated succinate, an oncometabolite that is proposed to drive tumorigenesis via DNA and histone hypermethylation, mitochondrial expansion, and pseudohypoxia-related gene expression. To interrogate this prevailing model, we disrupt mouse adrenal medulla SDHB expression, which recapitulates several key molecular features of human SDHx tumors, including succinate accumulation but not 5hmC loss, HIF accumulation, or tumorigenesis. By contrast, concomitant SDHB and the neurofibromin 1 tumor suppressor disruption yields SDHx-like pheochromocytomas. Unexpectedly, in vivo depletion of the 2-oxoglutarate (2-OG) dioxygenase cofactor ascorbate reduces SDHB-deficient cell survival, indicating that SDHx loss may be better tolerated by tissues with high antioxidant capacity. Contrary to the prevailing oncometabolite model, succinate accumulation and 2-OG-dependent dioxygenase inhibition are insufficient for mouse pheochromocytoma tumorigenesis, which requires additional growth-regulatory pathway activation.

Systematic Optimization of Ambient Ionization Ion Mobility Mass Spectrometry for Rapid Separation of Isomers.

Current methods typically used for metabolite screening and disease diagnosis often require extensive sample preparation, which increases analysis time and associated costs. While ambient ionization techniques enable the analysis of various samples in complex matrices with little or no sample preparation in a short time (typically within a minute), their reduced selectivity, even when coupled with high-resolution mass spectrometers, limits their application in certain fields. In this study, we have optimized the coupling of paper spray (PS) and leaf spray (LS) ambient ionization techniques with a commercially available ion mobility mass spectrometer (IM-MS) and demonstrated the separation of geometric and constitutional isomers. Ambient ionization techniques allow simultaneous introduction and ionization of samples, while background noise and matrix interference from paper and leaf substrates are filtered out by IM separation, resulting in high sensitivity and selectivity of the PS-IM-MS and LS-IM-MS workflows. In addition, we introduced a novel approach to perform single-field collision cross section (CCS) measurements, which resulted in CCS values that differ by 0.15% and 0.25% from traditional stepped-field and single-field methods, respectively. In addition, we used advanced computational tools to confidently identify analyte structures by comparing CCS values from experimental IM measurements and theoretical calculations. These results suggest that the coupling of ambient ionization methods with ion mobility techniques enables rapid, sensitive, and highly selective analysis that can be used in different fields, such as agrochemical screening and disease diagnostics.

Assessing the environmental ruggedness of paper spray ionization (PSI) coupled to a portable mass spectrometer operated under field conditions

Significant efforts have went towards coupling portable mass spectrometric systems and ambient ionization methods, with a culminating goal of field-based, point-of-need screening of harmful chemicals. While fieldable, ambient MS methods are an intriguing combination, ruggedness of these ionization methods in regard to naturally-occurring environmental conditions while operating in-situ, such as ambient temperature, relative humidity (RH) and wind, bears further investigation, as such reports are limited. In this work, a systematic characterization of paper spray ionization-mass spectrometry (PSI-MS) performance under variable environmental conditions was undertaken on a field-deployable MS system. Of all conditions, the presence of wind speeds greater than 3 m/s (∼7 MPH) was shown to have the most substantial effect on the spectral attributes of signal intensity and duration of the PSI-MS spraying event. Generally, moderate ambient temperature and %RH produced the best spectral performance for PSI-MS. Heat maps were generated from field-collected data across variable temperature and %RH in an effort to inform general PSI-MS performance trends and guide on-site operation by potential users.

Automated High-Throughput System Combining Small-Scale Synthesis with Bioassays and Reaction Screening.

The Purdue Make It system is a unique automated platform capable of small-scale in situ synthesis, screening small-molecule reactions, and performing direct label-free bioassays. The platform is based on desorption electrospray ionization (DESI), an ambient ionization method that allows for minimal sample workup and is capable of accelerating reactions in secondary droplets, thus conferring unique advantages compared with other high-throughput screening technologies. By combining DESI with liquid handling robotics, the system achieves throughputs of more than 1 sample/s, handling up to 6144 samples in a single run. As little as 100 fmol/spot of analyte is required to perform both initial analysis by mass spectrometry (MS) and further MSn structural characterization. The data obtained are processed using custom software so that results are easily visualized as interactive heatmaps of reaction plates based on the peak intensities of m/z values of interest. In this paper, we review the system's capabilities as described in previous publications and demonstrate its utilization in two new high-throughput campaigns: (1) the screening of 188 unique combinatorial reactions (24 reaction types, 188 unique reaction mixtures) to determine reactivity trends and (2) label-free studies of the nicotinamide N-methyltransferase enzyme directly from the bioassay buffer. The system's versatility holds promise for several future directions, including the collection of secondary droplets containing the products from successful reaction screening measurements, the development of machine learning algorithms using data collected from compound library screening, and the adaption of a variety of relevant bioassays to high-throughput MS.

Ambient Single-Cell Analysis and Native Tissue Imaging Using Laser-Ablation Electrospray Ionization Mass Spectrometry with Increased Spatial Resolution.

Laser-ablation electrospray ionization mass spectrometry (LAESI-MS) is an emerging method that has the potential to transform the field of metabolomics. This is in part due to LAESI-MS being an ambient ionization method that requires minimal sample preparation and uses (endogenous) water for in situ analysis. This application note details the employment of the "LAESI microscope" source to perform spatially resolved MS analysis of cells and MS imaging (MSI) of tissues at high spatial resolution. This source configuration utilizes a long-working-distance reflective objective that permits both visualization of the sample and a smaller LAESI laser beam profile than conventional LAESI setups. Here, we analyzed 200 single cells of Allium cepa (red onion) and imaged Fittonia argyroneura (nerve plant) in high spatial resolution using this source coupled to a Fourier transform mass spectrometer for high-mass-resolution and high-mass-accuracy metabolomics.

2D-Molybdenum Disulfide-Derived Ion Source for Mass Spectrometry.

Generation of current or potential at nanostructures using appropriate stimuli is one of the futuristic methods of energy generation. We developed an ambient soft ionization method for mass spectrometry using 2D-MoS2, termed streaming ionization, which eliminates the use of traditional energy sources needed for ion formation. The ionic dissociation-induced electrokinetic effect at the liquid-solid interface is the reason for energy generation. We report the highest figure of merit of current generation of 1.3 A/m2 by flowing protic solvents at 22 μL/min over a 1 × 1 mm2 surface coated with 2D-MoS2, which is adequate to produce continuous ionization of an array of analytes, making mass spectrometry possible. Weakly bound ion clusters and uric acid in urine have been detected. Further, the methodology was used as a self-energized breath alcohol sensor capable of detecting 3% alcohol in the breath.

Analysis of ion currents in mass spectrometric profiles using glioblastoma tissue

Background: The development of direct ambient ionization methods makes way for fast mass-spectrometry profiling of biological samples, which has great potential in medicine. Those methods, unlike traditional mass spectrometric analysis with chromatographic separation, are not able to take into account inter-ion interaction, ion suppression, and matrix effect due to the absence of chromatographic separation of the mixture components. So dynamics of ion current during direct ambient ionization mass-spectra is governed by the component micro-extraction and electrospray ionization influenced by the geometry of the sample, its position, and internal heterogeneity. Despite the progress in mass-spectrometry of biological samples, not much is known about the influence of sample type and structure on its molecular profile peculiarities. Methods: In this work, we propose to use analysis of the correlation between individual ion currents for a better understanding of ion current variability sources and grouping ions of high biological importance. Several fragments of glioblastoma tissue from a single patient are used for these purposes. Results: Ion currents have different dynamics considering different ions in different fragments. The correlation of two selected ion currents could be positive or negative for single fragment measurement. Correlations have persistent or alternating signs in different fragments for two selected ions. The spread of correlations of each pair of ion currents is calculated for evaluation of the signs’ stability. Conclusions: We were able to group ions according to the primary reason for their variabilities such as micro-extraction, mass-spectrometry measurement, or specimens' properties. Such grouping would allow the development of more reliable and reproducible methods of mass-spectrometry data analysis and improve the accuracy of results of its application in medicine.

Rapid detection of pesticide residues in Chinese herbal medicines by molecularly imprinted membrane electrospray ionization mass spectrometry

Chinese herbal medicines (CHMs) play an increasingly important role in the field of medicine and affects public health in the world. Although more and more strict has been employed to ensure the quality and safety of CHMs, pesticide residues in CHMs remain a serious issue and are the bottleneck for the global development of CHMs. In this work, we applied molecularly imprinted membrane electrospray mass spectrometry (MIM-ESI MS) for rapid detecting 4 classes of pesticide residues in CHMs, including organophosphorus (OPP), carbamates, pyrethroids and neonicotinoids in CHMs. Compared with our previous ambient ionization method MESI, MIM-ESI is capable of achieving a ∼50-fold increase in the detection limit of conventional analytical methods owing to the specificity recognition and unique enrichment of MIM. The optimal experimental conditions were determined, and the method was further validated for its sensitivity and specificity. Our data showed that MIM-ESI MS is applicable for the direct quantitation of pesticide residues in CHMs. This detection technology may help to ensure the quality of CHMs in the future.

2D MS/MS Spectra Recorded in the Time Domain Using Repetitive Frequency Sweeps in Linear Quadrupole Ion Traps.

Ion trap mass spectrometers have emerged as powerful on-site analytical platforms, in spite of limited mass resolution, due to their compatibility with ambient ionization methods and ready implementation of tandem mass spectrometry (MS/MS). When operated at constant trapping voltage, ions can be activated at their secular frequencies and all MS/MS experiments can be performed, including the two-dimensional tandem mass scan (2D MS/MS scan) in which all precursor ions and their subsequent product ions are both identified and correlated. In the new method of performing this 2D MS/MS experiment presented here, the precursor ions are excited by a nonlinear (inverse Mathieu q) frequency sweep while the resulting product ions are identified by their ejection time within a repeating orthogonally applied nonlinear (inverse Mathieu q) frequency sweep. This resulting compact representation contains the total fragmentation behavior of a collection of ionized compounds and captures detailed chemical information efficiently (typically in 1 s). The approach is implemented using a simple single mass analyzer instrument. This methodology was tested on three different multicomponent mixtures: drugs of abuse, peptides, and fentanyl analogs. The data are compared with those obtained by more common MS/MS scan methods.

Electroporation and Mass Spectrometry: A New Paradigm for In Situ Analysis of Intact Proteins Direct from Living Yeast Colonies.

Yeasts constitute an oft-neglected class of pathogens among which the resistance to first-line treatments, attributed in part to mutations in efflux pumps, is rapidly emerging. Their thick, chitin-reinforced cell walls render cell lysis difficult, complicating their analysis and identification by methods routinely used for bacteria, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Liquid extraction surface analysis mass spectrometry (LESA-MS) has previously been applied to the analysis of intact proteins from Gram-positive and Gram-negative bacterial colonies sampled directly on solid nutrient media. To date, a similar analysis of yeast colonies has not proved possible. Here we demonstrate the rapid release of intact yeast proteins for LESA-MS by electroporation using a home-built high-voltage device designed to lyse cells grown in colonies on agar media. Detection and identification of previously inaccessible proteins from baker’s yeast Saccharomyces cerevisiae, as well as two clinically relevant yeast species (Candida glabrata and Cryptococcus neoformans), is shown. The electroporation approach also has the potential to be translated to other mass spectrometric analysis techniques, including MALDI and various ambient ionization methods.

Natural Product Discovery by Direct Analysis in Real Time Mass Spectrometry.

Direct analysis in real time mass spectrometry (DART MS) is one of the first ambient ionization methods to be introduced and commercialized. Analysis by DART MS requires minimal sample preparation, produces nearly instantaneous results, and provides detection over a broad range of compounds. These advantageous features are particularly well-suited for the inherent complexity of natural product analysis. This review highlights recent applications of DART MS for species identification by chemotaxonomy, chemical profiling, genetic screening, and chemical spatial analysis from plants, insects, microbes, and metabolites from living systems.