Heterogeneous Oxidation Products of Fine Particulate Isoprene Epoxydiol-Derived Methyltetrol Sulfates Increase Oxidative Stress and Inflammatory Gene Responses in Human Lung Cells.

Hydroxyl radical (·OH)-initiated oxidation of isoprene, the most abundant nonmethane hydrocarbon in the atmosphere, is responsible for substantial amounts of secondary organic aerosol (SOA) within ambient fine particles. Fine particulate 2-methyltetrol sulfate diastereoisomers (2-MTSs) are abundant SOA products formed via acid-catalyzed multiphase chemistry of isoprene-derived epoxydiols with inorganic sulfate aerosols under low-nitric oxide conditions. We recently demonstrated that heterogeneous ·OH oxidation of particulate 2-MTSs leads to the particle-phase formation of multifunctional organosulfates (OSs). However, it remains uncertain if atmospheric chemical aging of particulate 2-MTSs induces toxic effects within human lung cells. We show that inhibitory concentration-50 (IC50) values decreased from exposure to fine particulate 2-MTSs that were heterogeneously aged for 0 to 22 days by ·OH, indicating increased particulate toxicity in BEAS-2B lung cells. Lung cells further exhibited concentration-dependent modulation of oxidative stress- and inflammatory-related gene expression. Principal component analysis was carried out on the chemical mixtures and revealed positive correlations between exposure to aged multifunctional OSs and altered expression of targeted genes. Exposure to particulate 2-MTSs alone was associated with an altered expression of antireactive oxygen species (ROS)-related genes (NQO-1, SOD-2, and CAT) indicative of a response to ROS in the cells. Increased aging of particulate 2-MTSs by ·OH exposure was associated with an increased expression of glutathione pathway-related genes (GCLM and GCLC) and an anti-inflammatory gene (IL-10).

Toxicological Responses of α-Pinene-Derived Secondary Organic Aerosol and Its Molecular Tracers in Human Lung Cell Lines.

Secondary organic aerosol (SOA) is a major component of airborne fine particulate matter (PM2.5) that contributes to adverse human health effects upon inhalation. Atmospheric ozonolysis of α-pinene, an abundantly emitted monoterpene from terrestrial vegetation, leads to significant global SOA formation; however, its impact on pulmonary pathophysiology remains uncertain. In this study, we quantified an increasing concentration response of three well-established α-pinene SOA tracers (pinic, pinonic, and 3-methyl-1,2,3-butanetricarboxylic acids) and a full mixture of α-pinene SOA in A549 (alveolar epithelial carcinoma) and BEAS-2B (bronchial epithelial normal) lung cell lines. The three aforementioned tracers contributed ∼57% of the α-pinene SOA mass under our experimental conditions. Cellular proliferation, cell viability, and oxidative stress were assessed as toxicological end points. The three α-pinene SOA molecular tracers had insignificant responses in both cell types when compared with the α-pinene SOA (up to 200 µg mL-1). BEAS-2B cells exposed to 200 µg mL-1 of α-pinene SOA decreased cellular proliferation to ∼70% and 44% at 24- and 48-h post exposure, respectively; no changes in A549 cells were observed. The inhibitory concentration-50 (IC50) in BEAS-2B cells was found to be 912 and 230 µg mL-1 at 24 and 48 h, respectively. An approximate 4-fold increase in cellular oxidative stress was observed in BEAS-2B cells when compared with untreated cells, suggesting that reactive oxygen species (ROS) buildup resulted in the downstream cytotoxicity following 24 h of exposure to α-pinene SOA. Organic hydroperoxides that were identified in the α-pinene SOA samples likely contributed to the ROS and cytotoxicity. This study identifies the potential components of α-pinene SOA that likely modulate the oxidative stress response within lung cells and highlights the need to carry out chronic exposure studies on α-pinene SOA to elucidate its long-term inhalation exposure effects.

Reaction Kinetics of Green Leaf Volatiles with Sulfate, Hydroxyl, and Nitrate Radicals in Tropospheric Aqueous Phase.

Green plants exposed to abiotic or biotic stress release C-5 and C-6 unsaturated oxygenated hydrocarbons called Green Leaf Volatiles (GLVs). GLVs partition into tropospheric waters and react to form secondary organic aerosol (SOA). We explored the kinetics of aqueous-phase reactions of 1-penten-3-ol (PENTOL), (Z)-2-hexen-1-ol (HEXOL), and (E)-2-hexen-1-al (HEXAL) with SO4•-, •OH, and NO3•. At 298 K, the rate constants for reactions of PENTOL, HEXOL, and HEXAL with SO4•- were, respectively, (9.4 ± 1.0) × 108 L mol-1 s-1, (2.5 ± 0.3) × 109 L mol-1 s-1, and (4.8 ± 0.2) × 108 L mol-1 s-1; with •OH - (6.3 ± 0.1) × 109 L mol-1 s-1, (6.7 ± 0.3) × 109 L mol-1 s-1, and (4.8 ± 0.3) × 109 L mol-1 s-1; and with NO3• - (1.5 ± 0.15) × 108 L mol-1 s-1, (8.4 ± 2.3) × 108 L mol-1 s-1, and (3.0 ± 0.7) × 107 L mol-1 s-1. The rate constants increased weakly with temperatures ranging from 278 to 318 K. The diffusional limitations of the rate constants appeared significant only for the GLV-•OH reactions. The aqueous-phase reactions appeared negligible in deliquescent aerosol and haze water but not in clouds and rains. The atmospheric lifetimes of GLVs decreased from many days to hours with increasing liquid water content and radicals' concentration.

Structural Characterization of Lactone-Containing MW 212 Organosulfates Originating from Isoprene Oxidation in Ambient Fine Aerosol.

Isoprene (C5H8) is the main non-methane hydrocarbon emitted into the global atmosphere. Despite intense research, atmospheric transformations of isoprene leading to secondary organic aerosol (SOA) are still not fully understood, including its multiphase chemical reactions. Herein, we report on the detailed structural characterization of atmospherically relevant isoprene-derived organosulfates (OSs) with a molecular weight (MW) of 212 (C5H8SO7), which are abundantly present in both ambient fine aerosol (PM2.5) and laboratory-generated isoprene SOA. The results obtained from smog chamber-generated isoprene SOA and aqueous-phase laboratory experiments coupled to the S(IV)-autooxidation chemistry of isoprene, 3-methyl-2(5H)-furanone, and 4-methyl-2(5H)-furanone, allowed us for the first time to fully elucidate the isomeric structures of the MW 212 OSs. By applying liquid chromatography interfaced to electrospray ionization high-resolution mass spectrometry, we firmly confirmed six positional isomers of the MW 212 OSs in PM2.5 collected from different sites in Europe and the United States. Our results also show that despite the low solubility of isoprene in water, aqueous-phase or multiphase chemistry can play an important role in the formation of OSs from isoprene. Possible formation mechanisms for the MW 212 OSs are also tentatively proposed.

Detection of Fungi and Oomycetes by Volatiles Using E-Nose and SPME-GC/MS Platforms.

Fungi and oomycetes release volatiles into their environment which could be used for olfactory detection and identification of these organisms by electronic-nose (e-nose). The aim of this study was to survey volatile compound emission using an e-nose device and to identify released molecules through solid phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS) analysis to ultimately develop a detection system for fungi and fungi-like organisms. To this end, cultures of eight fungi (Armillaria gallica, Armillaria ostoyae, Fusarium avenaceum, Fusarium culmorum, Fusarium oxysporum, Fusarium poae, Rhizoctonia solani, Trichoderma asperellum) and four oomycetes (Phytophthora cactorum, P. cinnamomi, P. plurivora, P. ramorum) were tested with the e-nose system and investigated by means of SPME-GC/MS. Strains of F. poae, R. solani and T. asperellum appeared to be the most odoriferous. All investigated fungal species (except R. solani) produced sesquiterpenes in variable amounts, in contrast to the tested oomycetes strains. Other molecules such as aliphatic hydrocarbons, alcohols, aldehydes, esters and benzene derivatives were found in all samples. The results suggested that the major differences between respective VOC emission ranges of the tested species lie in sesquiterpene production, with fungi emitting some while oomycetes released none or smaller amounts of such molecules. Our e-nose system could discriminate between the odors emitted by P. ramorum, F. poae, T. asperellum and R. solani, which accounted for over 88% of the PCA variance. These preliminary results of fungal and oomycete detection make the e-nose device suitable for further sensor design as a potential tool for forest managers, other plant managers, as well as regulatory agencies such as quarantine services.

Organic Hydroxy Acids as Highly Oxygenated Molecular (HOM) Tracers for Aged Isoprene Aerosol.

Highly oxygenated molecules (HOMs) are a class of compounds associated with secondary organic aerosols exhibiting high oxygen to carbon (O:C) ratios and often originating from the oxidation of biogenic compounds. Here, the photooxidation and ozonolysis of isoprene were examined under a range of conditions to identify HOM tracers for aged isoprene aerosol. The HOM tracers were identified as silylated derivatives by gas chromatography-mass spectrometry and by detecting their parent compounds by liquid chromatography-high resolution mass spectrometry. In addition to the previously observed methyltetrols and 2-methylglyceric acid, seven tracer compounds were identified, including 2-methyltartronic acid (MTtA), 2-methylerythronic acid (2MeTrA), 3-methylerythronic acid (3MeTrA), 2-methylthreonic acid (2MTrA), 3-methylthreonic acid (3MTrA), erythro-methyltartaric acid (e-MTA), and threo-methyltartaric acid (t-MTA). The molecular structures were confirmed with authentic standards synthesized in the laboratory. The presence of some of these HOMs in the gas and particle phases simultaneously provides evidence of their gas/particle partitioning. To determine the contributions of aged isoprene products to ambient aerosols, we analyzed ambient PM2.5 samples collected in the southeastern United States in summer 2003 and at two European monitoring stations located in Zielonka and Godów (Poland). Our findings show that methyltartaric acids (MTA) and 2- and 3-methylthreonic acids (and their stereoisomers) are representative of aged isoprene aerosol because they occur both in the laboratory chamber aerosol obtained and in ambient PM2.5. On the basis of gas chromatography-mass spectrometry (GC-MS) analysis, their concentrations were found to range from 0.04 ng for 3-methylthreonic acid to 6.3 ng m-3 for methyltartaric acid at the southeast site in Duke Forest, NC, USA.

Carbon-dot wrapped ZnO nanoparticle-based photoelectrochemical sensor for selective monitoring of H2O2 released from cancer cells.

This study reports on a simple approach for the fabrication of an electrode modified with biocompatible C-dot wrapped ZnO nanoparticles for selective photoelectrochemical monitoring of H2O2 released from living cells. The biocompatibility of the ZnO nanoparticles was confirmed through in-vitro cellular testing using the MTT assay on Huh7 cell lines. The ZnO nanoparticles wrapped with dopamine-derived C-dots possess numerous catalytically active sites, excessive surface defects, good electrical conductivity, and efficient separation ability of photo-induced electrons and holes. These properties offer highly sensitive and selective non-enzymatic photo-electrochemical monitoring of H2O2 released from HeLa cells after stimulation with N-formylmethionyl-leucyl-phenylalanine. The sensor has a wide linear range (20-800 nM), low detection limit (2.4 nM), and reliable reproducibility, this implying its suitability for biological and biomedical applications. Graphical abstract Schematic of the fabrication of ZnO nanoparticles by using a plant extract as a reducing agent. Wrapping of ZnO with C-dots enhances the photoelectrocatalytic efficacy. Sensitive and selective photoelectrochemical monitoring of H2O2 released from cancer cells is demonstrated.

Improved UHPLC-MS/MS Methods for Analysis of Isoprene-Derived Organosulfates.

Secondary organic aerosol (SOA) is an important yet not fully characterized constituent of atmospheric particulate matter. A number of different techniques and chromatographic methods are currently used for the analysis of SOA, so the comparison of results from different laboratories poses a challenge. So far, tentative structures have been suggested for many organosulfur compounds that have been identified as markers for the formation of SOA, including isoprene-derived organosulfates. Despite the effectiveness and robustness of LC-MS/MS analyses, the structural profiling of positional isomers of recently discovered organosulfates with molecular weights (MWs) of 214 and 212 from isoprene was entirely unsuccessful. Here, we developed a UHPLC combined with high-resolution tandem mass spectrometric method that significantly improves the separation efficiency and detection sensitivity of these compounds in aerosol matrices. We discovered that selection of the proper solvent for SOA extracts was a key factor in improving the separation parameters. Later, we took advantage of the enhanced sensitivity, combined with a short scan time window, to perform detailed structural mass-spectrometric studies. For the first time, we elucidate a number of isomers of the MW 214 and the MW 212 organosulfates and provide strong evidence for their molecular structures. The structure of trihydroxyketone sulfate MW 214 that we propose has not been previously reported. The methods we designed can be easily applied in other laboratories to foster an easy comparison of related qualitative and quantitative data obtained throughout the world.

Physical and Chemical Properties of 3-Methyl-1,2,3-butanetricarboxylic Acid (MBTCA) Aerosol.

The properties and the chemical fate of later generation products of the oxidation of biogenic organic compounds are mostly unknown. The properties of fresh MBTCA aerosol, a later generation product of the oxidation of monoterpenes in the atmosphere, were determined combining an aerosol mass spectrometer (AMS), a thermodenuder, and a scanning mobility particle sizer. Based on its AMS spectrum m/z 141.055 (C7H9O3+) could be used as an MBTCA signature. The MBTCA particle density was 1.43 ± 0.04 g cm-3, its saturation concentration was (1.8 ± 1.3) × 10-3 µg m-3 at 298 K, and its vaporization enthalpy was 150 ± 15 kJ mol-1. After OH radical exposure (∼1.2 days) and UV illumination the average aerosol O:C ratio decreased from 0.72 to 0.58-0.64 suggesting net fragmentation. Our findings suggest that the reactions of MBTCA with OH lead to CO2 loss with or without an oxygen addition.

Atmospheric aqueous phase radical chemistry of the isoprene oxidation products methacrolein, methyl vinyl ketone, methacrylic acid and acrylic acid--kinetics and product studies.

Kinetic and mechanistic studies were conducted on the isoprene oxidation products methacrolein, methyl vinyl ketone, methacrylic and acrylic acid reacting with hydroxyl and nitrate radicals and sulfate radical anions in aqueous solution by use of the laser flash photolysis technique and a reversed-rate method for kinetics. High-performance liquid chromatography/mass spectrometry was applied for product analysis. The kinetic investigations show the highest reactivity of the hydroxyl radical followed by sulfate and nitrate radicals. For methacrolein and methyl vinyl ketone the following rate constants have been determined at 298 K: k(OH+methacrolein) = (9.4 ± 0.7) × 10(9) M(-1) s(-1), k(OH+methyl vinyl ketone) = (7.3 ± 0.5) × 10(9) M(-1) s(-1), k(NO3+methacrolein) = (4.0 ± 1.0) × 10(7) M(-1) s(-1), k(NO3+methyl vinyl ketone) = (9.7 ± 3.4) × 10(6) M(-1) s(-1), k(SO4(-)+methacrolein) = (9.9 ± 4.9) × 10(7) M(-1) s(-1) and k(SO4(-)+methyl vinyl ketone) = (1.0 ± 0.2) × 10(8) M(-1) s(-1). Temperature and pH dependencies of the reactions of OH, NO3 and SO4(-) with methacrolein, methyl vinyl ketone, methacrylic and acrylic acid as well as Arrhenius parameters have been obtained and discussed. Product studies were performed on the OH radical induced oxidation of methacrolein and methyl vinyl ketone. In the reaction of methacrolein + OH methylglyoxal and hydroxyacetone were identified as first oxidation products with yields of 0.099 and 0.162. Methylglyoxal was primarily produced in the oxidation of methyl vinyl ketone with a yield of 0.052. For both precursor compounds the formation of glycolaldehyde was observed for the first time with yields of 0.051 and 0.111 in the oxidation of methacrolein and methyl vinyl ketone, respectively. Furthermore, highly functionalised C4 compounds were determined from the oxidation of both precursor compounds, but for the first time for methyl vinyl ketone. Reaction schemes were developed based on known peroxyl radical reaction mechanisms. The aqueous phase conversion of the first generation isoprene oxidation products can potentially contribute to tropospheric aqueous phase budgets of important carbonyl and dicarbonyl components which are expected to be conducive to the formation of aqSOA.

Dissociation constants of relevant secondary organic aerosol components in the atmosphere.

The presented studies focus on measuring the determination of the acidity constant (pKa) of relevant secondary organic aerosol components. For our research, we selected important oxidation products (mainly carboxylic acids) of the most abundant terpene compounds, such as α-pinene, ß-pinene, ß-caryophyllene, and δ-3-carene. The research covered the synthesis and determination of the acidity constant of selected compounds. We used three methods to measure the acidity constant, i.e., 1H NMR titration, pH-metric titration, Bates-Schwarzenbach spectrophotometric method. Moreover, the pKa values were calculated with Marvin 21.17.0 software to compare the experimentally derived values with those calculated from the chemical structure. pKa values measured with 1H NMR titration ranged from 3.51 ± 0.01 for terebic acid to 5.18 ± 0.06 for ß-norcaryophyllonic acid. Moreover, the data determined by the 1H NMR method revealed a good correlation with the data obtained with the commonly used potentiometric and UV-spectroscopic methods (R2 = 0.92). In contrast, the comparison with in silico results exhibits a relatively low correlation (R2Marvin = 0.66). We found that most of the values calculated with the Marvin Program are lower than experimental values obtained with pH-metric titration with an average difference of 0.44 pKa units. For di- and tricarboxylic acids, we obtained two and three pKa values, respectively. A good correlation with the literature values was observed, for example, Howell and Fisher (1958) used pH-metric titration and measured pKa1 and pKa2 to be 4.48 and 5.48, while our results are 4.24 ± 0.10 and 5.40 ± 0.02, respectively.

Evidence for cytotoxicity and mitochondrial dysfunction in human lung cells exposed to biomass burning aerosol constituents: Levoglucosan and 4-nitrocatechol.

Biomass burning (BB) emissions are one of the largest sources of carbonaceous aerosol, posing a significant risk as an airway irritant. Important BB markers include wood pyrolysis emissions, such as levoglucosan (LG) that is an anhydrous sugar bearing a six-carbon ring structure (i.e., 1,6-anhydro-ß-D-glucopyranose). Atmospheric chemical aging of BB-derived aerosol (BBA) in the presence of nitrogen oxides (NOx) can yield nitro-aromatic compounds, including 4-nitrocatechol (4NC). There is building evidence that NOx-mediated chemical aging of BBA poses a more serious exposure effect than primary pyrolysis emissions. This study provides a comparative toxicological assessment following the exposure to important BBA marker compounds in human lung cells (i.e., A549 and BEAS-2B) to determine whether aromatic 4NC is more toxic than BBA-bound anhydrous carbohydrate (i.e., LG). We determined inhibitory concentration-50 (IC50) and examined reactive oxygen species (ROS) changes, mitochondrial dysfunction, and apoptosis induction in the two cell lines following exposure to LG and 4NC in a dose-response manner. In the BEAS-2B cells, estimated IC50 values for 4NC were 33 and 8.8 µg mL-1, and for LG were 2546 and ∼3 × 107 µg mL-1 at 24 h and 48 h of exposure, respectively. A549 cells exhibited a much higher IC50 value than BEAS-2B cells. LG exposures resulted in mitochondrial stress with viability inhibition, but cells recovered with increasing exposure time. 4NC exposures at 200 µg mL-1 resulted in the induction of apoptosis at 6 h. Mitochondrial dysfunction and ROS imbalance induced the intrinsic apoptotic pathway induction following 4NC exposures. While increased ROS is caused by LG exposure in lung cells, 4NC is a marker of concern during BB emissions, as we observed apoptosis and high mitochondrial ROS in both lung cells at atmospherically-relevant aerosol concentrations. It may be associated with higher airway or inhalation pathologies in higher BBA emissions, such as wildfires or during wood combustion.

Aqueous-phase photo-oxidation of selected green leaf volatiles initiated by OH radicals: Products and atmospheric implications.

C5- and C6- unsaturated oxygenated organic compounds emitted by plants under stress like cutting, freezing or drying, known as Green Leaf Volatiles (GLVs), may clear some of the existing uncertainties in secondary organic aerosol (SOA) budget. The transformations of GLVs are a potential source of SOA components through photo-oxidation processes occurring in the atmospheric aqueous phase. Here, we investigated the aqueous photo-oxidation products from three abundant GLVs (1-penten-3-ol, (Z)-2-hexen-1-ol, and (E)-2-hexen-1-al) induced by OH radicals, carried out in a photo-reactor under simulated solar conditions. The aqueous reaction samples were analyzed using advanced hyphenated mass spectrometry techniques: capillary gas chromatography mass spectrometry (c-GC-MS); and reversed-phase liquid chromatography high resolution mass spectrometry (LC-HRMS). Using carbonyl-targeted c-GC-MS analysis, we confirmed the presence of propionaldehyde, butyraldehyde, 1-penten-3-one, and 2-hexen-1-al in the reaction samples. The LC-HRMS analysis confirmed the presence of a new carbonyl product with the molecular formula C6H10O2, which probably bears the hydroxyhexenal or hydroxyhexenone structure. Density functional theory (DFT)-based quantum calculations were used to evaluate the experimental data and obtain insight into the formation mechanism and structures of the identified oxidation products via the addition and hydrogen-abstraction pathways. DFT calculations highlighted the importance of the hydrogen abstraction pathway leading to the new product C6H10O2. Atmospheric relevance of the identified products was evaluated using a set of physical property data like Henry's law constant (HLC) and vapor pressure (VP). The unknown product of molecular formula C6H10O2 has higher HLC and lower VP than the parent GLV and thus has potential to remain in the aqueous phase leading to possible aqueous SOA formation. Other observed carbonyl products are likely first stage oxidation products and precursors of aged SOA.

Winter sources of PM2.5 pollution in Podkowa Lesna, a Central-European garden town (Mazovia, Poland).

The paper analyzes PM2.5 aerosol samples collected in Podkowa Lesna, a garden town in Mazovia, Central Poland, for 15 days in winter 2019. We determined the mass concentrations in the air of PM2.5 and PM2.5-bound organic carbon, elemental carbon, levoglucosan, and nine polycyclic aromatic hydrocarbons (PAHs). PM2.5 ranged from 11 to 51 µg m-3 (mean 31 µg m-3) and contained less than 32% organic carbon, 4% elemental carbon, 1% levoglucosan, and 0.12% total PAHs. The analysis based on positive matrix factorization (PMF) indicated two sources of PM2.5 of similar strength - burning vehicle fuels and biomass burning for residential heating. Levoglucosan originated exclusively from biomass burning, while 90% of elemental carbon was from vehicle emissions. About 62% of organic carbon, 85% of benzo(a)anthracene and 55-65% of the remaining PAHs originated from biomass burning. Compared to many worldwide locations, PM2.5, organic carbon, elemental carbon, and levoglucosan in Podkowa were among the lowest. The benzo(a)pyrene concentrations were the highest, while other PAHs were intermediate. However, the mass fractions of PAHs in Podkowa PM2.5 were the highest among the four locations available for comparison. That may indicate the low quality of fuel-burning processes. PAH-related inhalation cancer risk based on PAH carcinogenic potency in Podkowa appeared marginal. This work aims to induce local administrative actions to improve air quality in garden towns.

Newly Discovered Components of Dendrolimus pini Sex Pheromone.

The pine-tree lappet moth, D. pini, is a harmful defoliator of pine forests in Europe and Asia and a potentially invasive species in North America. The lures for trapping D. pini males based on two known components of its sex pheromone appeared weakly attractive to male moths. Identification of all components of the sex pheromone might allow for the development of more effective lures. The pheromone was sampled from virgin females using SPME and analyzed using gas chromatography coupled with mass spectrometry. Four new likely components ((Z5)-dodecenal, (Z5)-dodecen-1-ol, (Z5)-decen-1-yl acetate, (Z5)-tetradecen-1-yl acetate) and two known components ((Z5,E7)-dodecadienal, (Z5,E7)-dodecadien-1-ol) were identified based on comparison against authentic standards, Kováts indices and spectra libraries. The samples also contained several sesquiterpenes. Wind tunnel and field experiments showed that some blends of synthetic pheromone components alone or enriched with Scots pine essential oil (SPEO) were attractive to D. pini males. One component-(Z5)-decen-1-yl acetate-had a repelling effect. The presented knowledge of D. pini sex pheromone provides a basis for developing optimal lures for monitoring or controlling insect populations.

Cytotoxicity and oxidative stress induced by atmospheric mono-nitrophenols in human lung cells.

Nitrophenols (NPs) are hazardous pollutants found in various environmental matrices, including ambient fine particulate matter (PM2.5), agricultural residues, rainwater, wildfires, and industrial wastes. This study showed for the first time the effect of three pure nitrophenols and their mixture on human lung cells to provide basic understanding of the NP influence on cell elements and processes. We identified NPs in ambient PM2.5 and secondary organic aerosol (SOA) particles generated from the photooxidation of monocyclic aromatic hydrocarbons in the U.S. EPA smog chamber. We assessed the toxicity of identified NPs and their equimolar mixture in normal bronchial epithelial (BEAS-2B) and alveolar epithelial cancer (A549) lung cell lines. The inhibitory concentration-50 (IC50) values were highest and lowest in BEAS-2B cells treated with 2-nitrophenol (2NP) and 4-nitrophenol (4NP), respectively, at 24 h of exposure. The lactate dehydrogenase (LDH) assay showed that 4NP, the most abundant NP we identified in PM2.5, was the most cytotoxic NP examined in both cell lines. The annexin-V/fluorescein isothiocyanate (FITC) analysis showed that the populations of late apoptotic/necrotic BEAS-2B and A549 cells exposed to 3NP, 4NP, and NP equimolar mixture increased between 24 and 48 h. Cellular reactive oxygen species (ROS) buildup led to cellular death post exposure to 3NP, 4NP and the NP mixtures, while 2NP induced the lowest ROS buildup. An increased mitochondrial ROS signal following NP exposure occurred only in BEAS-2B cells. The tetramethylrhodamine, methyl ester, perchlorate (TMRM) assay showed that exposed cells exhibited collapse of the mitochondrial membrane potential. TMRM signals decreased significantly only in BEAS-2B cells, and most strongly with 4NP exposures. Our results suggest that acute atmospheric exposures to NPs may be toxic at high concentrations, but not at ambient PM2.5 concentrations. Further chronic studies with NP and NP-containing PM2.5 are warranted to assess their contribution to lung pathologies.

Toxicity of selected airborne nitrophenols on eukaryotic cell membrane models.

Nitroaromatics belong to the group of toxic components of aerosol particles and atmospheric hydrometeors that enter the atmosphere through biomass burning and fuel combustion. In the present work, we report on the cytotoxic effects of a 2-, 3- and 4-nitrophenol mixture on a model eukaryotic-like cell membrane and compared it with in vitro cellular models BEAS-2B (immortalized bronchial epithelial cells) and A549 (cancerous alveolar epithelial cells). A selected model biomembrane comprised of DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) was studied. The electrochemical-based method, combined with atomic force microscopy (AFM) and phase-contrast microscopy imaging, allowed to get insights into the mechanism of cellular function disruption caused by airborne nitrophenols. The efficacy of the method is supported by the data obtained from in vitro experiments performed on cell models. The nitrophenol mixture exhibited cytotoxic effects at concentrations above 100 µg mL-1, as demonstrated by phase-contrast microscopy in real lung cell lines. Electrochemical impedance spectroscopy (EIS) revealed the formation of membrane defects at a nitrophenol concentration of 200 µg mL-1. AFM imaging confirmed the model membrane disintegration and phospholipids rearrangement in the presence of nitrophenols. These observations indicate that particle-bound nitrophenols induce substantial changes in cell membranes and make them more permeable to aerosol, resulting in major cellular damage in the lungs when inhaled. The study provides initial evidence of cellular membrane damage induced by three important nitrated phenols present in the environment.

Rapid production of highly oxidized molecules in isoprene aerosol via peroxy and alkoxy radical isomerization pathways in low and high NOx environments: Combined laboratory, computational and field studies.

Recently, we identified seven novel hydroxy-carboxylic acids resulting from gas-phase reactions of isoprene in the presence of nitrogen oxides (NOx), ozone (O3), and/or hydroxyl radicals (OH). In the present study, we provide evidence that hydroxy-carboxylic acids, namely methyltartaric acids (MTA) are: (1) reliable isoprene tracers, (2) likely produced via rapid peroxy radical hydrogen atom (H) shift reactions (autoxidation mechanism) and analogous alkoxy radical H shifts in low and high NOx environments respectively and (3) representative of aged ambient aerosol in the low NOx regime. Firstly, MTA are reliable tracers of isoprene aerosol because they have been identified in numerous chamber experiments involving isoprene conducted under a wide range of conditions and are absent in the oxidation of mono- and sesquiterpenes. They are also present in numerous samples of ambient aerosol collected during the past 20 years at several locations in the U.S. and Europe. Furthermore, MTA concentrations measured during a year-long field study in Research Triangle Park (RTP), NC in 2003 show a seasonal trend consistent with isoprene emissions and photochemical activity. Secondly, an analysis of chemical ionization mass spectrometer (CIMS) data of several chamber experiments in low and high NOx environments show that highly oxidized molecules (HOMs) derived from isoprene that lead to MTAs may be produced rapidly and considered as early generation isoprene oxidation products in the gas phase. Density functional theory calculations show that rapid intramolecular H shifts involving peroxy and alkoxy radicals possess low barriers for methyl-hydroxy-butenals (MHBs) that may represent precursors for MTA. From these results, a viable rapid H shift mechanism is proposed to occur that produces isoprene derived HOMs like MTA. Finally, an analysis of the mechanism shows that autoxidation-like pathways in low and high NOx may produce HOMs in a few OH oxidation steps like commonly detected methyl tetrol (MT) isoprene tracers. The ratio of MTA/MT in isoprene aerosol is also shown to be significantly greater in field versus chamber samples indicating the importance of such pathways in the atmosphere even for smaller hydrocarbons like isoprene.

Tracing the biogenic secondary organic aerosol markers in rain, snow and hail.

The molecular characterization of secondary organic aerosol (SOA) is based mainly on LC-MS analyses of particulate matter (PM) samples collected with aerosol samplers. Several studies have analyzed atmospheric waters, including rain and cloud water, for the presence of SOA components, however, no separation techniques were used making identification of the individual components in these complex mixtures impossible. We have applied our improved UHPLC-HR-MS methodology to analyze atmospheric precipitates (hailstone, rain and snow), as well as SOA collected with high-volume samplers. We achieved sensitivity levels and separation efficiencies that were sufficient for molecular-level identification of individual compounds. Tracing commonly known SOA markers such as organosulfates (OS), C4-C6 dicarboxylic acids and terpenoic acids revealed that the chromatographic profiles for both atmospheric precipitate and PM samples were very similar, with both giving similar component ratios, especially for OS. We also demonstrated that SOA markers can be detected directly from raw rain samples. Our results show that LC-MS techniques are suitable for the convenient analysis of atmospheric precipitates containing SOA markers at the molecular level. It complements traditional SOA analyses and provides additional sampling opportunities which will no doubt allow for better elucidation of chemical transformations of volatile organic compounds in the atmosphere.