and technical progress achieved in FY16, as well as a list of selected publications and presentations that resulted from the research. For projects that concluded in FY16, a more detailed final report is provided that is technical in nature and includes the background, objectives, scientific approach, accomplishments, and impacts on the Laboratory missions, as well as a list of publications and presentations that resulted from the research. Project reports are listed under their research topics and organized by year and type, such as exploratory research (ER), feasibility study (FS), laboratory-wide competition (LW), and strategic initiative (SI). Each project is assigned a unique tracking code, an identifier that consists of three elements. The first is the fiscal year in which the project began, the second represents the project type, and the third identifies the serial number of the project for that fiscal year. For example, 16-ERD-100 means the project is an exploratory research project that began in FY16. The three-digit number (100) represents the serial number for the project.

and technical progress achieved in FY16, as well as a list of selected publications and presentations that resulted from the research. For projects that concluded in FY16, a more detailed final report is provided that is technical in nature and includes the background, objectives, scientific approach, accomplishments, and impacts on the Laboratory missions, as well as a list of publications and presentations that resulted from the research. Project reports are listed under their research topics and organized by year and type, such as exploratory research (ER), feasibility study (FS), laboratory-wide competition (LW), and strategic initiative (SI). Each project is assigned a unique tracking code, an identifier that consists of three elements. The first is the fiscal year in which the project began, the second represents the project type, and the third identifies the serial number of the project for that fiscal year. For example, 16-ERD-100 means the project is an exploratory research project that began in FY16. The three-digit number (100) represents the serial number for the project.

and technical progress achieved in FY16, as well as a list of selected publications and presentations that resulted from the research. For projects that concluded in FY16, a more detailed final report is provided that is technical in nature and includes the background, objectives, scientific approach, accomplishments, and impacts on the Laboratory missions, as well as a list of publications and presentations that resulted from the research. Project reports are listed under their research topics and organized by year and type, such as exploratory research (ER), feasibility study (FS), laboratory-wide competition (LW), and strategic initiative (SI). Each project is assigned a unique tracking code, an identifier that consists of three elements. The first is the fiscal year in which the project began, the second represents the project type, and the third identifies the serial number of the project for that fiscal year. For example, 16-ERD-100 means the project is an exploratory research project that began in FY16. The three-digit number (100) represents the serial number for the project.

At Lawrence Livermore National Laboratory, we have developed a single-particle aerosol mass spectrometry (SPAMS) system that can rapidly analyze individual micrometer-sized biological aerosol particles or cells that are sampled directly from air or a lab-generated aerosol into a mass spectrometer. As particles enter the SPAMS system, their aerodynamic size and fluorescence properties are measured before mass spectra from both positive and negative ions created by matrix-free laser desorption and ionization are recorded. All the correlated data obtained from a particle can be analyzed and classified in real-time. The SPAMS system is capable of discriminating, particle by particle, between bacterial spores, vegetative cells and other biological and non-biological background materials using the mass fingerprints obtained from those particles. In addition, selected species of bacteria can be discriminated from each other with this method. Here we describe the overall architecture of the SPAMS system and the related algorithms. We present selected results from applying the SPAMS technique to the analysis of biological agent simulants and single cells. We also describe results from first proof-of-concept experiments using SPAMS for the rapid screening of human effluents for tuberculosis. Lastly, we present results from a field study in a large airport using SPAMS to assess biological content in ambient aerosol.

The Bioaerosol Mass Spectrometry (BAMS) system was developed for the real-time detection and identification of biological aerosols using laser desorption ionization. Greater differentiation of particle types is desired; consequently MALDI techniques are being investigated. The small sample size (∼1 μm3), lack of substrate, and ability to simultaneously monitor both positive and negative ions provide a unique opportunity to gain new insight into the MALDI process. Several parameters known to influence MALDI molecular ion yield and formation are investigated here in the single particle phase. A comparative study of five matrices (2,6-dihydroxyacetophenone, 2,5-dihydroxybenzoic acid, α-cyano-4-hydroxycinnamic acid, ferulic acid, and sinapinic acid) with a single analyte (angiotensin I) is presented and reveals effects of matrix selection, matrix-to-analyte molar ratio, and aerosol particle diameter. The strongest analyte ion signal is found at a matrix-to-analyte molar ratio of 100:1. At this ratio, the matrices yielding the least and greatest analyte molecular ion formation are ferulic acid and α-cyano-4-hydroxycinnamic acid, respectively. Additionally, a significant positive correlation is found between aerodynamic particle diameter and analyte molecular ion yield for all matrices. SEM imaging of select aerosol particle types reveals interesting surface morphology and structure.

An in situ method to image individual aerosol particles upon desorption/ionization in a single particle aerosol mass spectrometer using shadowgraph imaging is presented and applied directly to determine a 2D spatial distribution of 5.0μm particles. This new technique is complementary to previously reported methods for the determination of focused particle beam profiles and has several advantages, including a very low particle concentration requirement and the ability to collect the image of an individual particle with a correlated mass spectrum. Images are presented for 1.3, 2.1, 3.0, and 5.0μm polystyrene latex spheres. Particle beam profiles are presented in the directions perpendicular and parallel to the particle beam axis. These results are qualitatively compared to those of a complementary particle collection method (dusting) used at Lawrence Livermore National Laboratory to characterize aerosol beam focusing.

Actual or surrogate chemical, biological, radiological, nuclear, and explosive materials and illicit drug precursors can be rapidly detected and identified when in aerosol form by a Single-Particle Aerosol Mass Spectrometry (SPAMS) system. This entails not only the sampling of such particles but also the physical analysis and subsequent data analysis leading to a highly reliable alarm state. SPAMS hardware is briefly reviewed. SPAMS software algorithms are discussed in greater detail. A laboratory experiment involving actual threat and surrogate releases mixed with ambient background aerosols demonstrates broad-spectrum detection within seconds. Data from a field test at the San Francisco International Airport demonstrate extended field operation with an ultralow false alarm rate. Together these data sets demonstrate a significant and important advance in rapid aerosol threat detection.

Single-particle aerosol mass spectrometry (SPAMS) was used for the real-time detection of liquid nerve agent simulants. A total of 1000 dual-polarity time-of-flight mass spectra were obtained for micrometer-sized single particles each of dimethyl methyl phosphonate, diethyl ethyl phosphonate, diethyl phosphoramidate, and diethyl phthalate using laser fluences between 0.58 and 7.83 nJ/microm2, and mass spectral variation with laser fluence was studied. The mass spectra obtained allowed identification of single particles of the chemical warfare agent (CWA) simulants at each laser fluence used although lower laser fluences allowed more facile identification. SPAMS is presented as a promising real-time detection system for the presence of CWAs.

The analysis of poly(ethylene glycol) (PEG)-containing particles by online single particle aerosol mass spectrometers equipped with laser desorption/ionization (LDI) is reported. We demonstrate that PEG-containing particles are useful in the development of aerosol mass spectrometers because of their ease of preparation, low cost, and inherently recognizable mass spectra. Solutions containing millimolar quantities of PEGs were nebulized and, after drying, the resultant micrometer-sized PEG-containing particles were sampled. LDI (266 nm) of particles containing NaCl and PEG molecules of average molecular weight<500 Da generated mass spectra reminiscent of mass spectra of PEG collected by other mass spectrometer platforms including the characteristic distribution of positive ions (Na+ adducts) separated by the 44 m/z units of the ethylene oxide units separating each degree of polymerization. PEGs of average molecular weight>500 Da were detected from particles that also contained the tripeptide tyrosine-tyrosine-tyrosine or 2,5-dihydroxybenzoic acid, which were added to nebulized solutions to act as matrices to assist LDI using pulsed 266 nm and 355 nm lasers, respectively. Experiments were performed on two aerosol mass spectrometers, one reflectron and one linear, that each utilize two time-of-flight mass analyzers to detect positive and negative ions created from a single particle. PEG-containing particles are currently being employed in the optimization of our bioaerosol mass spectrometers for the application of measurements of complex biological samples, including human effluents, and we recommend that the same strategies will be of great utility to the development of any online aerosol LDI mass spectrometer platform.

The application of single-particle aerosol mass spectrometry (SPAMS) to the real-time detection of micrometer-sized single particles of high explosives is described. Dual-polarity time-of-flight mass spectra from 1000 single particles each of 2,4,6-trinitrotoluene (TNT), 1,3,5-trinitro-1,3,5-triazinane (RDX), and pentaerythritol tetranitrate (PETN), as well as those of complex explosives, Composition B, Semtex 1A, and Semtex 1H, were obtained over a range of desorption/ionization laser fluences between 0.50 and 8.01 nJ/microm2. Mass spectral variability with laser fluence for each explosive is discussed. The ability of the SPAMS system to identify explosive components in a single complex explosive particle ( approximately 1 pg) without the need for consumables is demonstrated.

Bioaerosol Mass Spectrometry (BAMS), a real-time single cell analytical technique, was used to follow the biochemical and morphological changes within a group of Bacillus atrophaeus cells by measuring individual cells during the process of sporulation. A mutant of B. atrophaeus that lacks the ability to produce dipicolinic acid (DPA) was also analyzed. Single cell aerodynamic sizing was used to follow gross morphological changes, and chemical analysis of single cells by mass spectrometry was used to follow some biochemical changes of B. atrophaeus cells during endospore formation.

The Bio-Aerosol Mass Spectrometry (BAMS) system is an instrument used for the real time detection and identification of biological aerosols. Particles are drawn from the atmosphere directly into vacuum and tracked as they scatter light from several continuous wave lasers. After tracking, the fluorescence of individual particles is excited by a pulsed 266nm or 355nm laser. Molecules from those particles with appropriate fluorescence properties are subsequently desorbed and ionized using a pulsed 266nm laser. Resulting ions are analyzed in a dual polarity mass spectrometer. During two field deployments at the San Francisco International Airport, millions of ambient particles were analyzed and a small but significant fraction were found to have fluorescent properties similar to Bacillus spores and vegetative cells. Further separation of non-biological background particles from potential biological particles was accomplished using laser desorption/ionization mass spectrometry. This has been shown to enable some level of species differentiation in specific cases, but the creation and observation of higher mass ions is needed to enable a higher level of specificity across more species. A soft ionization technique, matrix-assisted laser desorption/ionization (MALDI) is being investigated for this purpose. MALDI is particularly well suited for mass analysis of biomolecules since it allows for the generation of molecular ions from large mass compounds that would fragment under normal irradiation. Some of the initial results from a modified BAMS system utilizing this technique are described.