Integrated data analysis making use of the total information from gas chromatography and high-resolution time-of-flight mass spectrometry to identify qualitative differences between two whisky samples.

RATIONALE: Analysis of complex mixtures with gas chromatography coupled with high-resolution time-of-flight mass spectrometry (GC/HRTOFMS) can produce a large amount of data. A new software program was recently reported that integrates all of the available mass spectrometric information from GC/HRTOFMS analysis into a concise report. New capabilities have now been added to the software to incorporate retention index data and to identify differences between two samples. METHODS: Two Scotch whisky samples were sampled by solid-phase microextraction (SPME) and analyzed by GC/HRTOFMS. One of the two whisky samples (Sample B) was identical to the other sample (Sample A) except for an additional 6-months storage in sherry casks. Both electron ionization (EI) and chemical ionization (CI) data were obtained using a new GC/TOFMS system (JEOL AccuTOF GC-Alpha) with enhanced resolving power and mass accuracy. Statistical analysis of replicate measurements of the whisky samples was carried out with a new software program (msFineAnalysis version 3.2) to identify and assign differences between the samples. RESULTS: There were 124 peaks detected in the two whiskies. Thirteen compounds were detected with a relative peak area greater than 0.05% that were present in greater amounts in the whisky that had been stored in sherry casks for an additional 6 months. Ten of these compounds were identified by the software with high confidence, two were identified as isomers with close retention indices, and one was identified interactively. CONCLUSIONS: The software identified small differences between the two samples that resulted from sherry cask aging. Because all of the information available from the hard and soft ionization analyses for each compound is summarized in a concise integrated report, the operator can rapidly determine the level of confidence for each assignment and inspect the information for compounds that are not present in the database.

Fast pesticide analysis using low-pressure gas chromatography paired with a triple quadrupole mass spectrometer equipped with short collision cell technology.

RATIONALE: A proof of concept showing GC-MS/MS analysis time for pesticides can be dramatically reduced while maintaining a similar separation efficiency by combining a low-pressure gas chromatography (LPGC) column with the enhanced selected reaction monitoring (SRM) switching speed of the short collision cell of a JEOL JMS-TQ4000GC. METHODS: Triple-quadrupole tandem mass spectrometry (standard EI + at 70 eV) was used to measure pesticides eluting from a low-pressure gas chromatograph capillary column. Three transitions for each of 244 pesticide compounds were measured within an 11-min analysis time, and the data were checked to confirm the method's reproducibility and ability to distinguish all three transitions for each pesticide. RESULTS: All three transitions for all 244 pesticides were detected in the standard mixture at 1X concentration within the 11-min analysis time. Relative standard deviation (RSD) of peak areas was less than 15% for 242 pesticides, and I/Q RSDs were less than 10% for 242 compounds. Retention time RSD over 15 replicates was less than 0.1%. CONCLUSIONS: Results show that analysis time can be markedly decreased using an LPGC column, and that the ability of the short collision cell to distinguish a large number of coeluting peaks makes the two technologies a natural pairing. The effective measurement of pesticides within a short time could benefit any scientists doing pesticide analysis work.

Cuticular hydrocarbons for the identification and geographic assignment of empty puparia of forensically important flies.

Research in social insects has shown that hydrocarbons on their cuticle are species-specific. This has also been proven for Diptera and is a promising tool for identifying important fly taxa in Forensic Entomology. Sometimes the empty puparia, in which the metamorphosis to the adult fly has taken place, can be the most useful entomological evidence at the crime scene. However, so far, they are used with little profit in criminal investigations due to the difficulties of reliably discriminate among different species. We analysed the CHC chemical profiles of empty puparia from seven forensically important blow flies Calliphora vicina, Chrysomya albiceps, Lucilia caesar, Lucilia sericata, Lucilia silvarum, Protophormia terraenovae, Phormia regina and the flesh fly Sarcophaga caerulescens. The aim was to use their profiles for identification but also investigate geographical differences by comparing profiles of the same species (here: C. vicina and L. sericata) from different regions. The cuticular hydrocarbons were extracted with hexane and analysed using gas chromatography-mass spectrometry. Our results reveal distinguishing differences within the cuticular hydrocarbon profiles allowing for identification of all analysed species. There were also differences shown in the profiles of C. vicina from Germany, Spain, Norway and England, indicating that geographical locations can be determined from this chemical analysis. Differences in L. sericata, sampled from England and two locations in Germany, were less pronounced, but there was even some indication that it may be possible to distinguish populations within Germany that are about 70 km apart from one another.

Rapid Fingerprinting of High-Molecular-Weight Polymers by Laser Desorption-Ionization Using Through-Hole Alumina Membrane High-Resolution Mass Spectrometry.

Residual acid found in the desorption ionization using through-holes alumina membranes (DIUTHAME) induces a reproducible protonation/in-source dissociation of polymers made of ester, amide, or siloxane moieties during their surface-assisted laser desorption ionization (SALDI) mass analysis. Deposited on the DIUTHAME chips in solution (solvent-based) or in pure form by melting the polymer powder in situ (solvent-free), high-molecular-weight nylons, silicone, or functionalized celluloses among other polymers are instantly fingerprinted by laser DIUTHAME high-resolution mass spectrometry (MS) with specific patterns resembling their direct analysis in real-time (DART) single-stage or tandem mass spectra. Depending on the polymer, two main types of fingerprints are observed with either the protonated monomer or product ions revealing the nature of the repeating unit or its functionalization. This technique allows a rapid molecular analysis of industrial homopolymers regardless of their molecular weight and complementary to DART with simple or no sample preparation and also promisingly applicable for copolymers.

Saccharomyces cerevisiae and S. pastorianus species and strain differentiation by direct analysis in real time time-of-flight mass spectrometry.

RATIONALE: Seventeen different dried yeast strains, including twelve strains of Saccharomyces cerevisiae and five strains of S. pastorianus, were analyzed using direct analysis in real time (DART) time-of-flight mass spectrometry. The resulting mass spectra were used for rapid species and strain differentiation based upon small-molecule metabolomic profiles. METHODS: Yeast strains purchased from local shops were suspended in a 1:1 water-methanol solution. Solutions were sampled by dipping the sealed end of a melting point capillary into each vial. Six replicates were measured in positive-ion and negative-ion mode for each strain using an automated linear rail with the DART source operated with helium gas and a gas heater temperature of 350°C. Averaged and centroided mass spectra were exported for analysis with chemometric software. RESULTS: Negative-ion DART mass spectra exhibited less chemical background and more distinctive components than positive-ion DART mass spectra. An on-line search of the Yeast Metabolome Database provided candidate metabolites for selection as features for chemometric analysis. Negative-ion DART mass spectra could distinguish both species and all strains. The DART analysis was also able to identify potential metabolomic differences between top-fermenting and bottom-fermenting yeast, between beer and baking yeast, and between red wine and champagne yeast. CONCLUSIONS: All strains could be distinguished by their negative-ion DART mass spectra with 97.7% validation accuracy. Clear differences were observed between dry and liquid forms and Saccharomyces strains with different applications to baking or beverage fermentation. Possible differences in metabolite profiles were suggested, but not confirmed, by accurate mass data.

Thermal desorption and pyrolysis direct analysis in real time mass spectrometry for qualitative characterization of polymers and polymer additives.

RATIONALE: Direct analysis in real time mass spectrometry (DART-MS) provides qualitative information about additives and polymer composition. However, the observed mass spectra are dependent on sampling conditions, in particular the DART gas temperature. This report describes the combination of a heated sample stage with DART-MS for polymer characterization. METHODS: Industrial polymers with different compositions were examined by thermal desorption and pyrolysis (TDPy) DART. Samples were heated on disposable copper stages from ambient temperature to 600°C, and the evolved gases were introduced directly into a DART ion source through a glass tee. Time- and temperature-dependent mass spectra were acquired using a high-resolution time-of-flight mass spectrometer. Kendrick mass analysis was applied to the interpretation of complex mass spectra observed for fluorinated polymers. RESULTS: Positive-ion DART mass spectra of common polymers exhibited peak series differing by monomer masses, often accompanied by a peak corresponding to the protonated monomer. Even polymers that did not exhibit a clear series of peaks produced characteristic mass spectra. Positive-ion and negative-ion mass spectra were recorded for fluorinated polymers, with polytetrafluoroethylene (PTFE) producing only negative ions. Thermal desorption provided characteristic temperature profiles for volatile species such as polymer additives and polymer pyrolysis products. CONCLUSIONS: In comparison with direct analysis by positioning sample directly in the heated DART gas stream, TDPy DART provides a more versatile sampling method and provides thermal separation and profiling of polymer additives, intact short polymer chains, and pyrolysis fragments.

Integrated qualitative analysis of polymer sample by pyrolysis-gas chromatography combined with high-resolution mass spectrometry: Using accurate mass measurement results from both electron ionization and soft ionization.

RATIONALE: Gas chromatography/mass spectrometry (GC/MS) is a powerful analytical tool used to separate and then identify volatile compounds through library database searches. However, as not all compounds are registered in these databases, it is not uncommon to detect unregistered components. Therefore, new analytical techniques were developed that utilize methods of identification beyond database searches alone. METHODS: Acquire data by using electron ionization (EI) and soft ionization (SI) with high-resolution mass spectrometry (HRMS). Use the EI mass spectra to library search for matches. Use the SI mass spectra for accurate mass analysis of the EI molecular ions. Conduct an isotope pattern analysis of the molecular ion to refine the possible candidate compositions. Use these compositions as a constraint for the accurate mass analysis of the EI fragment ions. If a given molecular ion formula is not correct, the EI fragment ions will not show good matches. Finally, all analytical results are integrated into a color-coded qualitative analysis report. RESULTS: The capabilities of this new integrated analytical method were assessed for a polymer resin sample that was measured by using pyrolysis-gas chromatography/high resolution time-of-flight mass spectrometry. A total of 161 compounds were detected in the total ion current chromatogram, and 154 of these compounds were identified as having only one chemical formula candidate with this new integrated qualitative analysis method. CONCLUSIONS: This new integrated qualitative analysis method gives analytical results independent of library search results. It can be applied to a variety of SI methods including chemical ionization, photoionization, field ionization, and low-energy EI.

Graphical Ranking of Divisors to Get the Most out of a Resolution-Enhanced Kendrick Mass Defect Plot.

Resolution-enhanced Kendrick Mass Defect (KMD) analysis using the new concept of fractional base units (repeating unit R divided by integer X; R/ X as a mathematical moiety) is now a powerful data-processing tool to unravel complex mass spectra of polymers. It enhances regular KMD analysis using the chemical moiety, R, to compute mass defects with unprecedented separation of ion series differing by their isotopic or comonomeric contents, end-groups, or charge states in highly visual KMD plots. The value of the divisor, X, dictates the gain of separating power from the regular to the resolution-enhanced KMD plot, and its choice strongly affects the ease and speed of data interpretation. A simple tool to help select the best values of X depending on the users' needs is mandatory to rationalize the analysis and avoid a time-consuming trial-and-error methodology. We propose two graphical representations intuitively ranking the well-suited divisors for the appropriate separation of isotopes or co-oligomers for copolymeric mass-spectral data. Rankings are extended to any type of data set from homopolymeric blends to terpolymers by generalizing the formulas with three variables beyond the specific separation of isotopes. The RANK functions are now available in commercial or homemade spreadsheets (available upon request) to interactively select divisors and compute the associated KMD plots.

Rapid paper spray mass spectrometry characterization of uranium and exemplar molecular species.

RATIONALE: The ability to detect and quantify the presence of specific inorganic elements and complexes is essential for environmental monitoring and nuclear safeguards applications. In this work, paper spray ionization mass spectrometry was used for the rapid chemical and isotopic characterization of trace inorganic species collected on cotton swipe substrates. The direct analysis of cotton swipes using this ambient ionization technique led to fast sample analysis that retained original chemical information of the source material with minimal sample preparation. METHODS: Mass spectra were collected with an atmospheric pressure ionization, high-resolution mass spectrometer for solutions containing uranyl acetate, uranyl chloride, uranyl nitrate, and uranyl tri-n-butylphosphate complexes. Gadolinium nitrate was used as an internal standard for the quantitative analysis of uranium. To demonstrate the ability to characterize inorganic contaminants in the presence of uranium, a multi-element inorganic standard containing U, Bi, Pb, Cd, Fe, and Zn was deposited onto cotton substrates and directly analyzed without purification. RESULTS: All elements doped on the cotton substrate were detected with strong signal-to-noise ratios (ca 1000 for UO2 + on multi-element doped swipes) and high integrated intensities (>105 counts) from collection periods of approximately 1 min. Limits of detection were determined to be approximately 94 ng for UO2 + and uranyl acetate through the measurement of ppb level solutions. CONCLUSIONS: The rapid analysis of uranium and other inorganic-containing samples while still retaining original chemical information (e.g. uranyl complexation) was demonstrated. Qualitative detection and speciation were achieved in less than 1 min of analysis. For uranium isotopic quantitation, longer accumulations (>15 min) can be sustained to improve the accuracy of minor 235 U isotopic abundance measurements to approximately 1% error.

Carbon-carbon double bond position elucidation in fatty acids using ozone-coupled direct analysis in real time mass spectrometry.

The carbon-carbon double bond positions of unsaturated fatty acids can have markedly different effects on biological function and also serve as biomarkers of disease pathology, dietary history, and species identity. As such, there is great interest in developing methods for the facile determination of double bond position for natural product chemistry, the pharmaceutical industry, and forensics. We paired ozonolysis with direct analysis in real time mass spectrometry (DART MS) to cleave and rapidly identify carbon-carbon double bond position in fatty acids, fatty alcohols, wax esters, and crude fatty acid extracts. In addition, ozone exposure time and DART ion source temperature were investigated to identify optimal conditions. Our results reveal that brief, offline exposure to ozone-generated aldehyde and carboxylate products that are indicative of carbon-carbon double bond position. The relative abundance of diagnostic fragments quantitatively reflects the ratios of isobaric fatty acid positional isomers in a mixture with a correlation coefficient of 0.99. Lastly, the unsaturation profile generated from unfractionated, fatty acid extracts can be used to differentiate insect species and populations. The ability to rapidly elucidate lipid double bond position by combining ozonolysis with DART MS will be useful for lipid structural elucidation, assessing isobaric purity, and potentially distinguishing between animals fed on different diets or belonging to different ecological populations.

"Reverse Kendrick Mass Defect Analysis": Rotating Mass Defect Graphs to Determine Oligomer Compositions for Homopolymers.

A new approach to determining the repeat unit compositions of homopolymers is reported in which a mass defect graph is rotated to zero slope to give a graph identical to a Kendrick mass defect graph. Because the Kendrick mass defect (KMD) is directly related to the elemental composition of the base unit, the process can be reversed. A mass defect graph (fractional m/ z plotted against exact m/ z) of a homopolymer can be rotated until the slope of the data points is zero. This is equivalent to finding a new constant factor by which the measured exact masses would have to be multiplied to create a Kendrick mass defect graph with zero slope. The elemental composition of the repeat unit can be determined by matching the new factor against the calculated factors for candidate compositions. This approach provides some benefits over simply looking for pairs of peaks corresponding to oligomer units. The primary benefit is to assist in visualization of the data. Rotating the data points corresponding to polymer masses to zero slope makes it easier to visualize the polymer data, and it facilitates the graphical isolation of polymer masses from background interferences. The repeat unit composition is determined not from a single pair of peaks but from multiple data points, and systematic errors in mass assignment can be visualized as deviations from linearity. Resolution-enhanced KMD graphs can be constructed for the calculated repeat unit composition by using fractional base units.

DART-MS: A New Analytical Technique for Forensic Paint Analysis.

Automotive paint evidence is one of the most significant forms of evidence obtained in automotive-related incidents. Therefore, the analysis of automotive paint evidence is imperative in forensic casework. Most analytical schemes for automotive paint characterization involve optical microscopy, followed by infrared spectroscopy and pyrolysis-gas chromatography mass spectrometry ( py-GCMS) if required. The main drawback with py-GCMS, aside from its destructive nature, is that this technique is relatively time intensive in comparison to other techniques. Direct analysis in real-time-time-of-flight mass spectrometry (DART-TOFMS) may provide an alternative to py-GCMS, as the rapidity of analysis and minimal sample preparation affords a significant advantage. In this study, automotive clear coats from four vehicles were characterized by DART-TOFMS and a standard py-GCMS protocol. Principal component analysis was utilized to interpret the resultant data and suggested the two techniques provided analogous sample discrimination. Moreover, in some instances DART-TOFMS was able to identify components not observed by py-GCMS and vice versa, which indicates that the two techniques may provide complementary information. Additionally, a thermal desorption/pyrolysis DART-TOFMS methodology was also evaluated to characterize the intact paint chips from the vehicles to ascertain if the linear temperature gradient provided additional discriminatory information. All the paint samples were able to be discriminated based on the distinctive thermal desorption plots afforded from this technique, which may also be utilized for sample discrimination. On the basis of the results, DART-TOFMS may provide an additional tool to the forensic paint examiner.

Development of "Laser Ablation Direct Analysis in Real Time Imaging" Mass Spectrometry: Application to Spatial Distribution Mapping of Metabolites Along the Biosynthetic Cascade Leading to Synthesis of Atropine and Scopolamine in Plant Tissue.

Methods for the accomplishment of small-molecule imaging by mass spectrometry are challenged by the need for sample pretreatment steps, such as cryo-sectioning, dehydration, chemical fixation, or application of a matrix or solvent, that must be performed to obtain interpretable spatial distribution data. Furthermore, these steps along with requirements of the mass analyzer such as high vacuum, can severely limit the range of sample types that can be analyzed by this powerful method. Here, we report the development of a laser ablation-direct analysis in real time imaging mass spectrometry approach which couples a 213 nm Nd:YAG solid state UV laser to a direct analysis in a real time ion source and high-resolution time-of-flight mass spectrometer. This platform enables facile determination of the spatial distribution of small-molecules spanning a range of polarities in a diversity of sample types and requires no matrix, vacuum, solvent, or complicated sample pretreatment steps. It furnishes high-resolution data, can be performed under ambient conditions on samples in their native form, and results in little to no fragmentation of analytes. We demonstrate its application through determination of the spatial distribution of molecules involved in the biosynthetic cascade leading to formation of the clinically relevant alkaloids atropine and scopolamine in Datura leichhardtii seed tissue.

Molecular Characterization of Volatiles and Petrochemical Base Oils by Photo-Ionization GC×GC-TOF-MS.

The characterization of organic mixtures by comprehensive two-dimensional gas chromatography (GC×GC) coupled to electron impact (EI) ionization time-of-flight mass spectrometry (TOF-MS) allows the detection of thousands of compounds. However, owing to the exhaustive fragmentation following EI ionization, despite the use of mass spectral libraries, a majority of the compounds remains unidentified because of the lack of parent ion preservation. Thus, soft-ionization energies leading to organic compounds being ionized with limited or no fragmentation, retaining the molecular ion, has been of interest for many years. In this study, photoionization (PI) was evaluated as the ion source for GC×GC-TOF-MS measurements. First, capabilities and limitations of PI were tested using an authentic mixture of compounds of several chemical classes. Ionization energy exhibited by PI, equivalent to 10.8 eV, resulted in significant retention of molecular ion information; [M]+• for alkanes, ketones, FAMEs, aromatics, [M-H]+• for chloroalkanes, and [M-H2O]+• for alcohols. Second, considering the potential of PI for hydrocarbons, base oils, complex mixtures of saturated and unsaturated hydrocarbons blended for finished lubricant formulations, were extensively evaluated. Several chemical classes of hydrocarbons were positively identified including a large number of isomeric compounds, both aliphatics and cyclics. Interestingly, branched-alkanes were ionized with lower excess internal energy, not only retaining the molecular ions but also exhibiting unique fragmentation patterns. The results presented herein offer a unique perspective into the detailed molecular characterization of base oils. Such unprecedented identification power of PI coupled with GC×GC-TOF-MS is the first report covering volatiles to low-volatile organic mixtures.

Mechanosensitivity below Ground: Touch-Sensitive Smell-Producing Roots in the Shy Plant Mimosa pudica.

The roots of the shy plant Mimosa pudica emit a cocktail of small organic and inorganic sulfur compounds and reactive intermediates into the environment, including SO2, methanesulfinic acid, pyruvic acid, lactic acid, ethanesulfinic acid, propanesulfenic acid, 2-aminothiophenol, S-propyl propane 1-thiosulfinate, phenothiazine, and thioformaldehyde, an elusive and highly unstable compound that, to our knowledge, has never before been reported to be emitted by a plant. When soil around the roots is dislodged or when seedling roots are touched, an odor is detected. The perceived odor corresponds to the emission of higher amounts of propanesulfenic acid, 2-aminothiophenol, S-propyl propane 1-thiosulfinate, and phenothiazine. The mechanosensitivity response is selective. Whereas touching the roots with soil or human skin resulted in odor detection, agitating the roots with other materials such as glass did not induce a similar response. Light and electron microscopy studies of the roots revealed the presence of microscopic sac-like root protuberances. Elemental analysis of these projections by energy-dispersive x-ray spectroscopy revealed them to contain higher levels of K(+) and Cl(-) compared with the surrounding tissue. Exposing the protuberances to stimuli that caused odor emission resulted in reductions in the levels of K(+) and Cl(-) in the touched area. The mechanistic implications of the variety of sulfur compounds observed vis-à-vis the pathways for their formation are discussed.

Rapid Species-level Identification of Salvias by Chemometric Processing of Ambient Ionisation Mass Spectrometry-derived Chemical Profiles.

INTRODUCTION: The Salvia genus contains numerous economically important plants that have horticultural, culinary and nutraceutical uses. They are often similar in appearance, making species determination difficult. Species identification of dried Salvia products is also challenging since distinguishing plant morphological features are no longer present. OBJECTIVE: The development of a simple high-throughput method of analysis of fresh and dried Salvia leaves that would permit rapid species-level identification and detection of diagnostic biomarkers. METHODOLOGY: Plant leaves were analysed in their native form by DART-MS without the need for any sample preparation steps. This furnished chemical fingerprints characteristic of each species. In the same experiment, in-source collision-induced dissociation was used to identify biomarkers. Biomarker presence was also independently confirmed by GC-MS. Chemometric processing of DART-MS profiles was performed by kernel discriminant analysis (KDA) and soft independent modelling of class analogy (SIMCA) to classify the fingerprints according to species. RESULTS: The approach was successful despite the occurrence of diurnal cycle and plant-age related chemical profile variations within species. In a single rapid experiment, the presence of essential oil biomarkers such as 3-carene, α-pinene, ß-pinene, ß-thujone, ß-caryophyllene, camphor and borneol could be confirmed. The method was applied to rapid identification and differentiation of Salvia apiana, S. dominica, S. elegans, S. officinalis, S. farinacea and S. patens. CONCLUSION: Species-level identification of Salvia plant material could be accomplished by chemometric processing of DART-HRMS-derived chemical profiles of both fresh and dried Salvia material. Copyright © 2016 John Wiley & Sons, Ltd.

Dopant-assisted direct analysis in real time mass spectrometry with argon gas.

RATIONALE: Dopants used with Atmospheric Pressure Photoionization (APPI) were examined with the Direct Analysis in Real Time (DART® ) ion source operated with argon gas. Charge-exchange and proton transfer reactions were observed by adding toluene, anisole, chlorobenzene and acetone to the DART gas stream, complementing the information obtained by helium DART. METHODS: Mass spectra were acquired with a time-of-flight mass spectrometer equipped with a DART ion source operated with argon gas. A syringe pump was used to introduce dopants directly into the DART gas stream through deactivated fused-silica capillary tubing. Samples including polycyclic aromatic hydrocarbons (PAHs), diesel fuel, trinitrotoluene and cannabinoids were deposited onto the sealed end of melting tube, allowed to dry, and the tube was then suspended in the dopant-enhanced DART gas stream. RESULTS: PAHs could be detected as molecular ions at concentrations in the low parts-per-billion range by using a solution of 0.5% anisole in toluene as a dopant. Argon DART analysis of a diesel fuel sample with the same dopant mixture showed a simpler mass spectrum than obtained by using helium DART. The argon DART mass spectrum was dominated by molecular ions for aromatic compounds, whereas the helium DART mass spectrum showed both molecular ions and protonated molecules. In contrast O2- attachment DART showed saturated hydrocarbons and oxygen-containing species. Mass spectra for trinitrotoluene with argon DART in negative-ion mode showed a prominent [M - H]- peak, whereas conventional helium DART showed both M- and [M - H]- . Lastly, in analogy to a report in the literature using APPI, positive ions produced by argon DART ionization for delta-9-tetrahydrocannabinol (THC) and cannabidiol showed distinctive product-ion mass spectra. CONCLUSIONS: Dopant-assisted argon DART operates by a mechanism that is analogous to those proposed for dopant-assisted atmospheric-pressure photoionization. Copyright © 2016 John Wiley & Sons, Ltd.

Alternative mass reference standards for direct analysis in real time mass spectrometry.

RATIONALE: Mass spectra were acquired with the Direct Analysis in Real Time (DART®) ion source for an amine-terminated polyether used as positive-ion mass reference standards and for several fluorinated materials commonly used as negative-ion reference standards for mass spectrometry. METHODS: A commercial time-of-flight mass spectrometer equipped with a DART ion source was used for all measurements. Mass reference standards deposited onto the sealed end of a glass melting point tube were suspended in the DART gas stream for analysis. RESULTS: A polyetheramine (Jeffamine® M-600) produced intense peaks corresponding to protonated molecules. Perfluorotributylamine (PFTBA), and perfluorotripentylamine, gave useful reference spectra for different m/z ranges. DART mass spectra of Ultramark 1621® resembled those previously reported for Fast Atom Bombardment (FAB) and Electrospray Ionization (ESI). Fomblin®Y, a fluorinated ether, was the most useful negative-ion reference standard of the materials tested. The material is commercially available, inexpensive, and provides reference peaks covering the m/z range 85 to >3000. CONCLUSIONS: Jeffamine-M600 was found to be a convenient alternative to polyethers such as polyethylene glycol (PEG) for DART positive-ion mass calibration. Fomblin Y was suitable for use as a negative-ion reference standard. Copyright © 2016 John Wiley & Sons, Ltd.

Plant seed species identification from chemical fingerprints: a high-throughput application of direct analysis in real time mass spectrometry.

Plant species identification based on the morphological features of plant parts is a well-established science in botany. However, species identification from seeds has largely been unexplored, despite the fact that the seeds contain all of the genetic information that distinguishes one plant from another. Using seeds of genus Datura plants, we show here that the mass spectrum-derived chemical fingerprints for seeds of the same species are similar. On the other hand, seeds from different species within the same genus display distinct chemical signatures, even though they may contain similar characteristic biomarkers. The intraspecies chemical signature similarities on the one hand, and interspecies fingerprint differences on the other, can be processed by multivariate statistical analysis methods to enable rapid species-level identification and differentiation. The chemical fingerprints can be acquired rapidly and in a high-throughput manner by direct analysis in real time mass spectrometry (DART-MS) analysis of the seeds in their native form, without use of a solvent extract. Importantly, knowledge of the identity of the detected molecules is not required for species level identification. However, confirmation of the presence within the seeds of various characteristic tropane and other alkaloids, including atropine, scopolamine, scopoline, tropine, tropinone, and tyramine, was accomplished by comparison of the in-source collision-induced dissociation (CID) fragmentation patterns of authentic standards, to the fragmentation patterns observed in the seeds when analyzed under similar in-source CID conditions. The advantages, applications, and implications of the chemometric processing of DART-MS derived seed chemical signatures for species level identification and differentiation are discussed.

Identification of bacteria by fatty acid profiling with direct analysis in real time mass spectrometry.

RATIONALE: Bacterial fatty acid profiling is a well-established technique for bacterial identification. Current methods involving esterification and gas chromatography/mass spectrometry (GC/MS) or matrix-assisted laser desorption/ionization (MALDI) analysis are effective, but there are potential benefits to be gained by investigating ambient ionization methods that can provide rapid analysis without derivatization or additional sample handling. METHODS: Lipid extracts from colonies of five Gram-positive and five Gram-negative pathogenic bacteria were analyzed by Direct Analysis in Real Time (DART) ionization coupled with a time-of-flight mass spectrometer. Fatty acid profiles were obtained from the negative-ion DART mass spectra without additional derivatization or sample preparation. RESULTS: Fatty acid profiles obtained from the deprotonated molecules [M - H](-) were found to be highly species-specific and reproducible. Leave-one-out cross validation (LOOCV) for principal component analysis (PCA) showed 100% correct classification accuracy. CONCLUSIONS: The results of this preliminary feasibility study show good precision and accuracy, and the fatty acid patterns are clearly distinctive for each of the ten species examined. The speed and ease of analysis and the high classification accuracy for this initial study indicate that DART is an effective method for bacterial fatty acid profiling.