Secondary ionization of chemical warfare agent simulants: atmospheric pressure ion mobility time-of-flight mass spectrometry

For the first time, the use of a traditional ionization source for ion mobility spectrometry (radioactive nickel ((63)Ni) beta emission ionization) and three alternative ionization sources (electrospray ionization (ESI), secondary electrospray ionization (SESI), and electrical discharge (corona) ionization (CI)) were employed with an atmospheric pressure ion mobility orthogonal reflector time-of-flight mass spectrometer (IM(tof)MS) to detect chemical warfare agent (CWA) simulants from both aqueous- and gas-phase samples. For liquid-phase samples, ESI was used as the sample introduction and ionization method. For the secondary ionization (SESI, CI, and traditional (63)Ni ionization) of vapor-phase samples, two modes of sample volatilization (heated capillary and thermal desorption chamber) were investigated. Simulant reference materials, which closely mimic the characteristic chemical structures of CWA as defined and described by Schedule 1, 2, or 3 of the Chemical Warfare Convention treaty verification, were used in this study. A mixture of four G/V-type nerve simulants (dimethyl methylphosphonate, pinacolyl methylphosphonate, diethyl phosphoramidate, and 2-(butylamino)ethanethiol) and one S-type vesicant simulant (2-chloroethyl ethyl sulfide) were found in each case (sample ionization and introduction methods) to be clearly resolved using the IM(tof)MS method. In many cases, reduced mobility constants (K(o)) were determined for the first time. Ion mobility drift times, flight times, relative signal intensities, and fragmentation product signatures for each of the CWA simulants are reported for each of the methods investigated.     

Environmentally acceptable sorbents of chemical warfare agent simulants

Development of environmentally acceptable decontaminants of chemical weapons and other highly toxic chemicals is important because of security, economical, health, political, and environmental reasons. The efficiency of natural zeolite (clinoptilolite) and synthetic zeolite, metal oxides, and their mixtures as chemical and physical sorbents of chemical warfare agents (CWA) was investigated. Commonly studied chemical warfare agent simulants dimethyl methylphosphonate (DMMP) and 2-chloroethyl ethyl sulfide (2-CEES) were included in this study. Organic solutions of DMMP and 2-CEES were passed through a column filled with natural and synthetic zeolites and their mixtures with metal oxides. After passing through the column filled with different sorbents, all eluents were filtered and centrifuged before the gas chromatography–mass spectroscopy (GC–MS) analysis. The efficiency of investigated adsorbents was estimated based on the obtained data. All investigated sorbents exhibited absorption efficiency for both simulants of chemical warfare agents. Infrared spectroscopy was used for the detection of DMMP and 2-CEES adsorbed to the investigated adsorbents. Since GC–MS analysis results indicate very good sorption properties of both simulants, the detection of adsorbed CWA simulants was a matter of routine.     

Detection of a chemical warfare agent simulant in various aerosol matrixes by ion mobility time-of-flight mass spectrometry

For the first time, a traditional radioactive nickel (63Ni) beta emission ionization source for ion mobility spectrometry was employed with an atmospheric pressure ion mobility orthogonal reflector time-of-flight mass spectrometer (IM(tof)MS) to detect a chemical warfare agent (CWA) simulant from aerosol samples. Aerosol-phase sampling employed a quartz cyclonic chamber for sample introduction. The simulant reference material, which closely mimicked the characteristic chemical structure of CWAs as defined and described by Schedule 1, 2, or 3 of the Chemical Warfare Convention treaty verification, was used in this study. An overall elevation in arbitrary signal intensity of approximately 1.0 orders of magnitude was obtained by the progressive increase of the thermal AP-IMS temperature from 75 to 275 degrees C. A mixture of one G-type nerve simulant (dimethyl methylphosphonate (DMMP)) in four (water, kerosene, gasoline, diesel) matrixes was found in each case (AP-IMS temperature 75-275 degrees C) to be clearly resolved in less than 2.20 x 10(4) micros using the IM(tof)MS instrument. Corresponding ions, masses, drift times, K(o) values, and arbitrary signal intensities for each of the sample matrixes are reported for the CWA simulant DMMP.     

Degradation of chemical warfare agent simulants using gas-liquid pulsed streamer discharges

This study determines the effectiveness of pulsed streamer discharges (PSD), a type of advanced oxidation technology (AOT) to clean water contaminated with chemical agents. For the purpose of this study, experiments were conducted with G and H agent simulants to determine the degradation kinetics and to determine the effects of various electrical and chemical parameters in the degradation of these contaminants. The energy efficiency of contaminant degradation shows that pulsed streamer discharges can be an efficient technology in treating water contaminated with chemical agents. The maximum energy yields of degradation of H and G agent simulants by the pulsed corona discharges are 0.029 and 0.008 molecules/100 eV, respectively, in the series configuration with ferrous sulfate salt in solution.     

Two-dimensional nonlinear wavelet compression of ion mobility spectra of chemical warfare agent simulants

Ion mobility spectrometry (IMS) affords miniaturized hand-held devices that can be used for monitoring and remote measurement. Because such instruments have limits on storage capacity or bandwidth for wireless transmission, data compression is important. Furthermore, all instruments should be operated with the fastest possible sampling rates because a signal-to-noise gain can be achieved by wavelet compression. Linear wavelet compression (LWC) applied to IMS data may cause peak distortion when the spectra are reconstructed. Nonlinear wavelet compression (NLWC) precisely preserves the peak location (i.e., drift time), height, and shape. IMS data of three chemical warfare simulants, dimethyl methylphosphonate, triethyl phosphate, and dipropyleneglycol monomethyl ether, were collected from an Ion Track ITEMISER and a Graseby Ionics detector CAM. Two-dimensional NLWC was used to compress the IMS data in the drift time and data acquisition dimensions on IMS data of chemical warfare simulants. NLWC was applied to achieve a compression factor of 1/128 with relative error of root-mean-square of <0.25% in the reconstructed spectra. A method was also developed and evaluated for optimizing compression.     

Surface-enhanced Raman scattering detection of chemical and biological agent simulants

Surface-enhanced Raman scattering spectra of chemical and biological agent simulants, such as dimethyl methylphonate, pinacolyl methylphosphonate, diethyl phosphoramidate, 2-chloroethyl ethylsulfide, bacillus globigii, erwinia herbicola, and bacillus thuringiensis were obtained from silver-oxide film-deposited substrates. Thin AgO films ranging in thickness from 50 to 250 nm were produced by chemical bath deposition onto glass slides. Further Raman intensity enhancements were noticed in UV irradiated surfaces due to photo-induced Ag nanocluster formation, which may provide a possible route to producing highly useful plasmonic sensors for the detection of chemical and biological agents upon visible-light illumination.     

Classification of chemical and biological warfare agent simulants by surface-enhanced Raman spectroscopy and multivariate statistical techniques

Initial results demonstrating the ability to classify surface-enhanced Raman (SERS) spectra of chemical and biological warfare agent simulants are presented. The spectra of two endospores (B. subtilis and B. atrophaeus), two chemical agent simulants (dimethyl methylphosphonate (DMMP) and diethyl methylphosphonate (DEMP)), and two toxin simulants (ovalbumin and horseradish peroxidase) were studied on multiple substrates fabricated from colloidal gold adsorbed onto a silanized quartz surface. The use of principal component analysis (PCA) and hierarchical clustering were used to evaluate the efficacy of identifying potential threat agents from their spectra collected on a single substrate. The use of partial least squares-discriminate analysis (PLS-DA) and soft independent modeling of class analogies (SIMCA) on a compilation of data from separate substrates, fabricated under identical conditions, demonstrates both the feasibility and the limitations of this technique for the identification of known but previously unclassified spectra.     

Detection of chemical weapon agents and simulants using chemical ionization reaction time-of-flight mass spectrometry

Chemical ionization reaction time-of-flight mass spectrometry (CIR-TOF-MS) has been used for the analysis of prepared mixtures of chemical weapon agents (CWAs) sarin and sulfur mustard. Detection of the CWA simulants 2-chloroethyl ethyl sulfide, triethyl phosphate, and dimethyl methyl phosphonate has also been investigated. Chemical ionization of all the agents and simulants was shown to be possible using the CIR-TOF-MS technique with a variety of reagent ions, and the sensitivity was optimized by variation of instrument parameters. The ionization process was found to be largely unaffected by sample humidity levels, demonstrating the potential suitability of the method to a range of environmental conditions, including the analysis of CWAs in air and in the breath of exposed individuals.     

Rapid screening of aqueous chemical warfare agent degradation products: ambient pressure ion mobility mass spectrometry

The use of electrospray ionization ambient pressure ion mobility spectrometry with an orthogonal reflector time-of-flight mass spectrometer to analyze chemical warfare (CW) degradation products from aqueous environmental samples has been demonstrated. Certified reference materials of analytical standards for the detection of Schedule 1, 2, or 3 toxic chemicals or their precursors as defined by the chemical warfare convention treaty verification were used in this study. A combination of six G/V-type nerve and four S-type vesicant related CW agent degradation products were separated with baseline resolution by this instrumental technique. Analytical figures of merit for each CW degradation product were determined. In some cases, reduced mobility constants (K0) have been reported for the first time. linear response ranges for the selected CW degradation products were found to be generally approximately 2 orders of magnitude, where the overall dynamic response ranges were found to extend to 4 orders of magnitude. Limits of detection for five of the nine chemical products tested were found to be less than 1 ppm. To demonstrate the potential of this instrumental method with complex mixtures, four CW degradation products were separated and detected from a spiked Palouse River water sample in less than 1 min. Finally, a homologous series of n-alkylamines were used as baseline reference standards, producing a mobility/mass trend line to which the CW degradation products could be compared. Comparison of these products in this manner is expected to reduce the number of false positive/negative responses.     

SIMPLISMA and ALS applied to two-way nonlinear wavelet compressed ion mobility spectra of chemical warfare agent simulants

Ion mobility spectrometry is a rapid scanning measurement method for which compression methods that facilitate the handling of large collections of data are beneficial. Peak distortion in reconstructed ion mobility spectra from linear wavelet compression is problematic in that artifact peaks may cause false positive alarms. Peak shifting also may cause false alarms if target peaks shift out of or interfering peaks shift into detection windows. Nonlinear wavelet compression (NLWC) preserves peak shape and can lessen the degree of distortion, shifting, and artifact peaks in the reconstructed spectra. NLWC was applied to achieve high compression and fidelity in the reconstructed spectra. Another benefit is that NLWC improves signal-to-noise ratios and thus the models built from compressed data are improved. By compressing both the drift time order and the spectrum acquisition order, greater compressions maybe achieved. A two-way nonlinear wavelet compression method that incorporates alternating least squares (2W-NLWC-ALS) algorithm was devised by applying ALS to partially reconstructed wavelet coefficients generated from two-way NLWC. The number of components in a data set can be determined automatically using ASIMPLISMA. The smaller ALS models are saved as the final compressed data and can be used to reconstruct the entire data set efficiently without maintaining the compressed wavelet coefficient matrix of the original data set. The 2W-NLWC-ALS algorithm provides greater compression ratios compared to regular wavelet compression and interpretable models. Using this method, large volumes of data can be acquired and easily evaluated through a simple compressed model. A compression ratio of 510 ppm, root-mean-square error (E(RMS)) of 6.3 mV (full-scale signal is usually 1 V or larger), and relative root-mean-square error (RE(RMS)) of 1.62% were achieved for data sets collected by CAM. A compression ratio of 46 ppm, E(RMS) of 9.2 mV, and RE(RMS) of 0.42% were achieved for data sets collected with an ITEMISER instrument. The 2W-NLWC-ALS algorithm is an efficient compression method that provides the benefits of a simple model.     

Enhancing the detection of sulfate particles for laser ablation aerosol mass spectrometry

A major limitation to the application of laser ablation aerosol mass spectrometry for the detection of particles less than 200 nm in diameter is a low ablation efficiency for sulfate particles. (Ablation efficiency is the probability that an ablated particle produces a detectable ion signal.) A method is described here to enhance the ablation efficiency of sulfate particles by coating them with a UV-absorbing compound. The method can be applied in-line with the aerosol mass spectrometer in a manner that does not significantly alter the aerosol size distribution. It is shown that a 12-nm coating of 1-naphthyl acetate increases the ablation efficiency of 136-nm ammonium sulfate particles by at least a factor of 20, while similar coatings on oleic acid and ammonium nitrate particles do not significantly alter the ablation efficiency. The results suggest that "undetected" particles, presumably sulfate, in ambient aerosol can be assessed.     

Multiplexed liquid arrays for simultaneous detection of simulants of biological warfare agents

Liquid array-based multiplexed immunoassays designed for rapid, sensitive, specific, and simultaneous detection of multiple simulants of biological warfare agents have been developed. In both blind and standard laboratory trials, we demonstrate the simultaneous detection of four simulant agents from a single sample. The challenge agents comprise broad classes of pathogens (virus, protein toxins, bacterial spores, vegetative cells). Assay performance of each analyte was optimized, and dose-response curves and the limits of detection (LODs) for individual analytes are presented. Assay performance, including dynamic range, sensitivity, and LODs for liquid arrays and enzyme-linked immunosorbant assay were compared and are shown to be similar. Maximum assay sensitivity is obtained in approximately 1 h, and good sensitivity is achieved in as little as 30 min. Although the sample matrixes are very complex, even for highly multiplexed assays the samples do not exhibit evidence of nonspecific binding, demonstrating that the assays also have high specificity.     

Ion mobility spectrometry and its applications in detection of chemical warfare agents

When fast detection of chemical warfare agents in the field is required, the ion mobility spectrometer may be the only suitable option. This article provides an essential survey of the different ion mobility spectrometry detection technologies. (To listen to a podcast about this feature, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html.).     

Measured infrared optical cross sections for a variety of chemical and biological aerosol simulants

We conducted a series of spectral extinction measurements on a variety of aerosolized chemical and biological simulants over the spectral range 3-13 microm using conventional Fourier-transform IR (FTIR) aerosol spectroscopy. Samples consist of both aerosolized particulates and atomized liquids. Materials considered include Bacillus subtilis endospores, lyophilized ovalbumin, polyethylene glycol, dimethicone (SF-96), and three common background materials: kaolin clay (hydrated aluminum silicate), Arizona road dust (primarily SiO2), and diesel soot. Aerosol size distributions and mass density were measured simultaneously with the FTIR spectra. As a result, all optical parameters presented here are mass normalized, i.e., in square meters per gram. In an effort to establish the utility of using Mie theory to predict such parameters, we conducted a series of calculations. For materials in which the complex indices of refraction are known, e.g., silicone oil (SF-96) and kaolin, measured size distributions were convolved with Mie theory and the resultant spectral extinction calculated. Where there was good agreement between measured and calculated extinction spectra, absorption, total scattering, and backscatter were also calculated.     

Single-particle mass spectrometry of polystyrene microspheres and diamond nanocrystals.

High-resolution mass spectra of single submicrometer-sized particles are obtained using an electrospray ionization source in combination with an audio frequency quadrupole ion-trap mass spectrometer. Distinct from conventional methods, light scattering from a continuous Ar-ion laser is detected for particles ejected out of the ion trap. Typically, 10 particles are being trapped and interrogated in each measurement. With the audio frequency ion trap operated in a mass-selective instability mode, analysis of the particles reveals that they all differ in mass-to-charge ratio (m/z), and the individual peak in the observed mass spectrum is essentially derived from one single particle. A histogram of the spectra acquired in 10(2) repetitions of the experiment is equivalent to the single spectrum that would be observed when an ion ensemble of 10(3) particles is analyzed simultaneously using the single-particle mass spectrometer (SPMS). To calibrate such single-particle mass spectra, secular frequencies of the oscillatory motions of the individual particle within the trap are measured, and the trap parameter qz at the point of ejection is determined. A mass resolution exceeding 10(4) can readily be achieved in the absence of ion ensemble effect. We demonstrate in this work that the SPMS not only allows investigations of monodisperse polystyrene microspheres, but also is capable of detecting diamond nanoparticles with a nominal diameter of 100 nm, as well.     

Quantitative detection of aromatic compounds in single aerosol particle mass spectrometry

Most laser-based aerosol mass spectrometers rely on a single ultraviolet laser to both ablate and ionize the aerosol particle. This technique produces complex and fragmented mass spectra, especially for organic compounds. The approach presented here achieves a more robust and quantitative analysis using a CO2 laser to evaporate the aerosol particle and a vacuum ultraviolet laser to ionize the vapor plume. Vacuum ultraviolet laser ionization produces little fragmentation in the mass spectra, making the identification of an aerosol particle's constituents more straightforward. An analysis of simple, three-component mixtures of aniline, benzyl alcohol, and m-nitrotoluene shows that the technique also provides a quantitative analysis for all the components of the mixture. Furthermore, the detection of predominantly parent ion signal from anthracene particles demonstrates the utility of the technique in the analysis of lower vapor pressure, solid-phase aerosols. Finally, we discuss the potential and limitations of this technique in analyzing organic atmospheric aerosols.     

Fiber-optic microsphere-based arrays for multiplexed biological warfare agent detection

We report a multiplexed high-density DNA array capable of rapid, sensitive, and reliable identification of potential biological warfare agents. An optical fiber bundle containing 6000 individual 3.1-mum-diameter fibers was chemically etched to yield microwells and used as the substrate for the array. Eighteen different 50-mer single-stranded DNA probes were covalently attached to 3.1-mum microspheres. Probe sequences were designed for Bacillus anthracis, Yersinia pestis, Francisella tularensis, Brucella melitensis, Clostridium botulinum, Vaccinia virus, and one biological warfare agent (BWA) simulant, Bacillus thuringiensis kurstaki. The microspheres were distributed into the microwells to form a randomized multiplexed high-density DNA array. A detection limit of 10 fM in a 50-microL sample volume was achieved within 30 min of hybridization for B. anthracis, Y. pestis, Vaccinia virus, and B. thuringiensis kurstaki. We used both specific responses of probes upon hybridization to complementary targets as well as response patterns of the multiplexed array to identify BWAs with high accuracy. We demonstrated the application of this multiplexed high-density DNA array for parallel identification of target BWAs in spiked sewage samples after PCR amplification. The array's miniaturized feature size, fabrication flexibility, reusability, and high reproducibility may enable this array platform to be integrated into a highly sensitive, specific, and reliable portable instrument for in situ BWA detection.     

In situ infrared aerosol spectroscopy for a variety of nerve agent simulants using flow-through photoacoustics

We present newly measured results of an ongoing experimental program established to measure optical cross sections in the mid- and long-wave infrared for a variety of chemically and biologically based aerosols. For this study we consider only chemically derived aerosols, and in particular, a group of chemical compounds often used as simulants for the detection of extremely toxic organophosphorus nerve agents. These materials include: diethyl methylphosphonate (DEMP), dimethyl methylphosphonate (DMMP), diisopropyl methylphosphonate (DIMP), and diethyl phthalate (DEP). As reported in a prior study [Appl. Opt. 44, 4001 (2005)], we combine two optical techniques well suited for aerosol spectroscopy [i.e., flow-through photoacoustics and Fourier transform infrared (FTIR) emission spectroscopy], to measure in situ the absolute extinction and absorption cross sections over a variety of wavelengths spanning the IR spectral region from 3 to 13 mum. Aerosol size distribution(s), particle number density, and dosimetric measurements are recorded simultaneously in order to present optical cross sections that are aerosol mass normalized, i.e., m(2)/gram. Photoacoustic results, conducted at a series of CO(2) laser lines, compare well with measured broadband FTIR spectral extinction. Both FTIR and photoacoustic data also compare well with Mie theory calculations based on measured size distributions and previously published complex indices of refraction.     

Simultaneous detection and identification of O-GlcNAc-modified glycoproteins using liquid chromatography-tandem mass spectrometry

Glycoproteins carrying O-linked N-acetylglucosamine (O-GlcNAc) modifications have been isolated from a wide range of organisms ranging from trypanosomes to humans. Interest in this modification is increasing as evidence accumulates that it is an abundant and transient modification that is dynamic and responsive to cellular stimuli. Concurrent advances in biological mass spectrometry (MS) have facilitated high-sensitivity protein identification by tandem MS. In this study, we show that the lability of the O-GlcNAc moiety to low-energy collision in tandem MS offers a means of distinguishing such peptides from others that are not modified. The differential between the energy required to remove the O-GlcNAc group and the energy required to fragment the peptide chain allows the O-GlcNAc group to be detected and the peptide sequence, and therefore the protein, to be identified. This technique thus allows the simultaneous detection and identification of O-GlcNAc-modified peptides, even when present at low levels in complex mixtures. The method was initially developed and validated using a synthetic O-GlcNAc-modified peptide and then applied to the detection of an extremely low abundance O-GlcNAc-modified peptide from bovine alpha-crystallin. We believe that with further development this assay system may prove to be a useful tool for the direct investigation of intracellular O-GlcNAc levels, thus providing valuable insights into the physiological role of O-GlcNAc modified proteins.     

Enhanced detection and identification of glycopeptides in negative ion mode mass spectrometry

A combined mass spectrometry (MS) and tandem mass spectrometry (MS/MS) approach implemented with matrix-assisted laser desorption ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI FTICR MS) in the negative ion mode is described for enhanced glycopeptide detection and MS/MS analysis. Positive ion mode MS analysis is widely used for glycopeptide characterization, but the analyses are hampered by potential charge-induced fragmentation of the glycopeptides and poor detection of the glycopeptides harboring sialic acids. Furthermore, tandem MS analysis (MS/MS) via collision-induced dissociation (CID) of glycopeptides in the positive ion mode predominantly yields glycan fragmentation with minimal information to verify the connecting peptide moiety. In this study, glycoproteins such as, bovine lactoferrin (b-LF) for N-glycosylation and kappa casein (k-CN) for O-glycosylation were analyzed in both the positive- and negative ion modes after digestion with bead-immobilized Pronase. For the b-LF analysis, 44 potential N-linked glycopeptides were detected in the positive ion mode while 61 potential N-linked glycopeptides were detected in the negative ion mode. By the same token, more O-linked glycopeptides mainly harboring sialic acids from k-CN were detected in the negative ion mode. The enhanced glycopeptide detection allowed improved site-specific analysis of protein glycosylation and superior to positive ion mode detection. Overall, the negative ion mode approach is aimed toward enhanced N- and O-linked glycopeptide detection and to serve as a complementary tool to positive ion mode MS/MS analysis.