Monique Williams

Metal and silicate particles including nanoparticles are present in electronic cigarette cartomizer fluid and aerosol.

Background Electronic cigarettes (EC) deliver aerosol by heating fluid containing nicotine. Cartomizer EC combine the fluid chamber and heating element in a single unit. Because EC do not burn tobacco, they may be safer than conventional cigarettes. Their use is rapidly increasing worldwide with little prior testing of their aerosol. Objectives We tested the hypothesis that EC aerosol contains metals derived from various components in EC. Methods Cartomizer contents and aerosols were analyzed using light and electron microscopy, cytotoxicity testing, x-ray microanalysis, particle counting, and inductively coupled plasma optical emission spectrometry. Results The filament, a nickel-chromium wire, was coupled to a thicker copper wire coated with silver. The silver coating was sometimes missing. Four tin solder joints attached the wires to each other and coupled the copper/silver wire to the air tube and mouthpiece. All cartomizers had evidence of use before packaging (burn spots on the fibers and electrophoretic movement of fluid in the fibers). Fibers in two cartomizers had green deposits that contained copper. Centrifugation of the fibers produced large pellets containing tin. Tin particles and tin whiskers were identified in cartridge fluid and outer fibers. Cartomizer fluid with tin particles was cytotoxic in assays using human pulmonary fibroblasts. The aerosol contained particles >1 µm comprised of tin, silver, iron, nickel, aluminum, and silicate and nanoparticles (<100 nm) of tin, chromium and nickel. The concentrations of nine of eleven elements in EC aerosol were higher than or equal to the corresponding concentrations in conventional cigarette smoke. Many of the elements identified in EC aerosol are known to cause respiratory distress and disease. Conclusions The presence of metal and silicate particles in cartomizer aerosol demonstrates the need for improved quality control in EC design and manufacture and studies on how EC aerosol impacts the health of users and bystanders.

Conventional and electronic cigarettes (e-cigarettes) have different smoking characteristics

INTRODUCTION Electronic cigarettes or e-cigarettes are marketed as tobacco-free nicotine delivery devices that have received little laboratory evaluation. In this study, the smoking properties of conventional and e-cigarettes were compared by examining the vacuum required to produce smoke (conventional cigarettes) or aerosol (e-cigarettes) and the density of the smoke/aerosol over time. METHODS Vacuum was measured using a manometer coupled to a smoking machine. The density of aerosol or smoke was measured spectrophotometrically. E-cigarettes were subjected to smoke-out experiments in which vacuum and aerosol density were measured until each cartridge was exhausted. RESULTS The vacuum required to smoke conventional cigarettes varied among the eight brands tested. Lights and ultra-light brands required stronger vacuums to smoke than unfiltered and regular filtered brands. Except for one brand, higher vacuums were required to smoke e-cigarettes than conventional brands. Smoke/aerosol density was stable for conventional brands and for e-cigarettes over the first 10 puffs; however, aerosol density of e-cigarettes dropped during subsequent smoking, and higher vacuums were required to produce aerosol as the puff number increased. While conventional cigarettes were uniform in their smoking behavior within brands, vacuum and density varied within brands of e-cigarettes. DISCUSSION Generally, e-cigarettes required stronger vacuums (suction) to smoke than conventional brands, and the effects of this on human health could be adverse. The amount of aerosol produced by e-cigarettes decreased during smoking, which necessitated increasing puff strength to produce aerosol. The decreased efficiency of aerosol production during e-cigarette smoking makes dosing nonuniform over time and calls into question their usefulness as nicotine delivery devices.

Variability Among Electronic Cigarettes in the Pressure Drop, Airflow Rate, and Aerosol Production

INTRODUCTION This study investigated the performance of electronic cigarettes (e-cigarettes), compared different models within a brand, compared identical copies of the same model within a brand, and examined performance using different protocols. METHODS Airflow rate required to generate aerosol, pressure drop across e-cigarettes, and aerosol density were examined using three different protocols. RESULTS First 10 puff protocol: The airflow rate required to produce aerosol and aerosol density varied among brands, while pressure drop varied among brands and between the same model within a brand. Total air hole area correlated with pressure drop for some brands. Smoke-out protocol: E-cigarettes within a brand generally performed similarly when puffed to exhaustion; however, there was considerable variation between brands in pressure drop, airflow rate required to produce aerosol, and the total number of puffs produced. With this protocol, aerosol density varied significantly between puffs and gradually declined. CONSECUTIVE TRIAL PROTOCOL: Two copies of one model were subjected to 11 puffs in three consecutive trials with breaks between trials. One copy performed similarly in each trial, while the second copy of the same model produced little aerosol during the third trial. The different performance properties of the two units were attributed to the atomizers. CONCLUSION There was significant variability between and within brands in the airflow rate required to produce aerosol, pressure drop, length of time cartridges lasted, and production of aerosol. Variation in performance properties within brands suggests a need for better quality control during e-cigarette manufacture.

Strategies to Reduce Tin and Other Metals in Electronic Cigarette Aerosol.

Background Metals are present in electronic cigarette (EC) fluid and aerosol and may present health risks to users. Objective The objective of this study was to measure the amounts of tin, copper, zinc, silver, nickel and chromium in the aerosol from four brands of EC and to identify the sources of these metals by examining the elemental composition of the atomizer components. Methods Four brands of popular EC were dissected and the cartomizers were examined microscopically. Elemental composition of cartomizer components was determined using integrated energy dispersive X-ray microanalysis, and the concentrations of the tin, copper, zinc silver, nickel, and chromium in the aerosol were determined for each brand using inductively coupled plasma optical emission spectroscopy. Results All filaments were made of nickel and chromium. Thick wires were copper coated with either tin or silver. Wires were joined to each other by tin solder, brazing, or by brass clamps. High concentrations of tin were detected in the aerosol when tin solder joints were friable. Tin coating on copper wires also contributed to tin in the aerosol. Conclusions Tin concentrations in EC aerosols varied both within and between brands. Tin in aerosol was reduced by coating the thick wire with silver rather than tin, placing stable tin solder joints outside the atomizing chamber, joining wires with brass clamps or by brazing rather than soldering wires. These data demonstrate the feasibility of removing tin and other unwanted metals from EC aerosol by altering designs and using materials of suitable quality.

Elements including metals in the atomizer and aerosol of disposable electronic cigarettes and electronic hookahs.

Objective Our purpose was to quantify 36 inorganic chemical elements in aerosols from disposable electronic cigarettes (ECs) and electronic hookahs (EHs), examine the effect of puffing topography on elements in aerosols, and identify the source of the elements. Methods Thirty-six inorganic chemical elements and their concentrations in EC/EH aerosols were determined using inductively coupled plasma optical emission spectroscopy, and their source was identified by analyzing disassembled atomizers using scanning electron microscopy and energy dispersive X-ray spectroscopy. Results Of 36 elements screened, 35 were detected in EC/EH aerosols, while only 15 were detected in conventional tobacco smoke. Some elements/metals were present in significantly higher concentrations in EC/EH aerosol than in cigarette smoke. Concentrations of particular elements/metals within EC/EH brands were sometimes variable. Aerosols generated at low and high air-flow rates produced the same pattern of elements, although the total element concentration decreased at the higher air flow rate. The relative amount of elements in the first and last 60 puffs was generally different. Silicon was the dominant element in aerosols from all EC/EH brands and in cigarette smoke. The elements appeared to come from the filament (nickel, chromium), thick wire (copper coated with silver), brass clamp (copper, zinc), solder joints (tin, lead), and wick and sheath (silicon, oxygen, calcium, magnesium, aluminum). Lead was identified in the solder and aerosol of two brands of EHs (up to 0.165 μg/10 puffs). Conclusion These data show that EC/EH aerosols contain a mixture of elements, including heavy metals, with concentrations often significantly higher than in conventional cigarette smoke. While the health effects of inhaling mixtures of heated metals is currently not known, these data will be valuable in future risk assessments involving EC/EH elements/metals.

Disposable Electronic Cigarettes and Electronic Hookahs: Evaluation of Performance

INTRODUCTION The purpose of this study was to characterize the performance of disposable button-activated and disposable airflow-activated electronic cigarettes (EC) and electronic hookahs (EH). METHODS The airflow rate required to produce aerosol, pressure drop, and the aerosol absorbance at 420 nm were measured during smoke-outs of 9 disposable products. Three units of each product were tested in these experiments. RESULTS The airflow rates required to produce aerosol and the aerosol absorbances were lower for button-activated models (3 mL/s; 0.41-0.55 absorbance) than for airflow-activated models (7-17 mL/s; 0.48-0.84 absorbance). Pressure drop was also lower across button-activated products (range = 6-12 mm H2O) than airflow-activated products (range = 15-67 mm H20). For 25 of 27 units tested, airflow did not have to be increased during smoke-out to maintain aerosol production, unlike earlier generation models. Two brands had uniform performance characteristics for all parameters, while 3 had at least 1 product that did not function normally. While button-activated models lasted 200 puffs or less and EH airflow-activated models often lasted 400 puffs, none of the models produced as many puffs as advertised. Puff number was limited by battery life, which was shorter in button-activated models. CONCLUSION The performance of disposable products was differentiated mainly by the way the aerosol was produced (button vs airflow-activated) rather than by product type (EC vs EH). Users needed to take harder drags on airflow-activated models. Performance varied within models, and battery life limited the number of puffs. Data suggest quality control in manufacturing varies among brands.

NEIL2 protects against oxidative DNA damage induced by sidestream smoke in human cells.

Secondhand smoke (SHS) is a confirmed lung carcinogen that introduces thousands of toxic chemicals into the lungs. SHS contains chemicals that have been implicated in causing oxidative DNA damage in the airway epithelium. Although DNA repair is considered a key defensive mechanism against various environmental attacks, such as cigarette smoking, the associations of individual repair enzymes with susceptibility to lung cancer are largely unknown. This study investigated the role of NEIL2, a DNA glycosylase excising oxidative base lesions, in human lung cells treated with sidestream smoke (SSS), the main component of SHS. To do so, we generated NEIL2 knockdown cells using siRNA-technology and exposed them to SSS-laden medium. Representative SSS chemical compounds in the medium were analyzed by mass spectrometry. An increased production of reactive oxygen species (ROS) in SSS-exposed cells was detected through the fluorescent detection and the induction of HIF-1α. The long amplicon–quantitative PCR (LA-QPCR) assay detected significant dose-dependent increases of oxidative DNA damage in the HPRT gene of cultured human pulmonary fibroblasts (hPF) and BEAS-2B epithelial cells exposed to SSS for 24 h. These data suggest that SSS exposure increased oxidative stress, which could contribute to SSS-mediated toxicity. siRNA knockdown of NEIL2 in hPF and HEK 293 cells exposed to SSS for 24 h resulted in significantly more oxidative DNA damage in HPRT and POLB than in cells with control siRNA. Taken together, our data strongly suggest that decreased repair of oxidative DNA base lesions due to an impaired NEIL2 expression in non-smokers exposed to SSS would lead to accumulation of mutations in genomic DNA of lung cells over time, thus contributing to the onset of SSS-induced lung cancer.

iQOS: evidence of pyrolysis and release of a toxicant from plastic

Objective To evaluate performance of the I quit original smoking (iQOS) heat-not-burn system as a function of cleaning and puffing topography, investigate the validity of manufacturer’s claims that this device does not burn tobacco and determine if the polymer-film filter is potentially harmful. Methods iQOS performance was evaluated using five running conditions incorporating two different cleaning protocols. Heatsticks were visually and stereomicroscopically inspected preuse and postuse to determine the extent of tobacco plug charring (from pyrolysis) and polymer-film filter melting, and to elucidate the effects of cleaning on charring. Gas chromatography–mass spectrometry headspace analysis was conducted on unused polymer-film filters to determine if potentially toxic chemicals are emitted from the filter during heating. Results For all testing protocols, pressure drop decreased as puff number increased. Changes in testing protocols did not affect aerosol density. Charring due to pyrolysis (a form of organic matter thermochemical decomposition) was observed in the tobacco plug after use. When the manufacturer’s cleaning instructions were followed, both charring of the tobacco plug and melting of the polymer-film filter increased. Headspace analysis of the polymer-film filter revealed the release of formaldehyde cyanohydrin at 90°C, which is well below the maximum temperature reached during normal usage. Discussion Device usage limitations may contribute to decreases in interpuff intervals, potentially increasing user’s intake of nicotine and other harmful chemicals. This study found that the tobacco plug does char and that charring increases when the device is not cleaned between heatsticks. Release of formaldehyde cyanohydrin is a concern as it is highly toxic at very low concentrations.

Comparison of the Performance of Cartomizer Style Electronic Cigarettes from Major Tobacco and Independent Manufacturers

Objective This study compared the performance of 12 brands of cartomizer style electronic cigarettes (EC) using different puffing protocols and measured the concentrations of nicotine in each product. Methods Air flow rate, pressure drop, and aerosol absorbance were measured using two different protocols, first 10 puffs and a modified smoke-out protocol. Results First 10 puff protocol: The air flow rate required to produce aerosol ranged between brands from 4–21 mL/s. Pressure drop was relatively stable within a brand but ranged between brands from 14–71 mmH2O and was much lower than the earlier classic 3-piece models. Absorbance, a measure of aerosol density, was relatively consistent between puffs, but varied between brands. With the modified smoke-out protocol, most brands were puffed until 300 puffs. The pressure drop was relatively stable for all brands except three. Absorbance of the aerosol decreased as the number of puffs increased. Although there was some uniformity in performance within some brands, there was large variation between brands. The labeled and measured nicotine concentrations were within 10% of each other in only 1 out of 10 brands. Conclusions Over 10 puffs, the cartomizers all perform similarly within a brand but varied between brands. In smoke-out trials, most brands lasted at least 300 puffs, and performed similarly within brands with respect to pressure drop and absorbance. For five brands, products purchased at different times performed differently. These data show some improvement in performance during evolution of these products, but nevertheless indicate problems with quality control in manufacture.

Aberrant Cell and Basement Membrane Architecture Contribute to Sidestream Smoke-induced Choroidal Endothelial Dysfunction

PURPOSE Environmental tobacco smoke (ETS) is widely regarded as a major modifiable risk factor for age-related macular degeneration (AMD). Yet, precisely how it exerts its pathologic effects is poorly understood. Since early-stage AMD is characterized by choroidal capillary loss, this study examined the effect of sidestream smoke (SS), the major component of ETS, on the viability of choroidal endothelial cells (EC), with an emphasis on the role of aberrant cell and basement membrane (BM) architecture in mediating SS-induced response. METHODS Chorioretinal ECs (RF/6A) were treated with SS, and cell viability and architecture were analyzed by colorimetric assay and actin cytoskeletal organization, respectively. The structure of RF/6A EC-secreted BM was examined by immunofluorescence for collagen IV and immunoblotting for lysyl oxidase (LOX), a collagen-crosslinking enzyme. Finally, fresh RF/6A ECs were cultured on decellularized SS-treated BM to evaluate its active role in EC dysfunction. RESULTS The RF/6A EC viability decreased progressively with increasing SS dose, which correlated strongly with a significant decline in actin cytoskeleton-dependent EC spreading. Sidestream smoke also caused marked disruption of the RF/6A EC-secreted BM that was accompanied by suppression of LOX expression. Further, fresh, non-SS-treated RF/6A ECs exhibited a significant loss in viability and actin cytoskeletal organization when cultured on SS-treated corrupt BM. CONCLUSIONS These findings indicate that aberrant physical cues in the form of EC and BM architecture likely have an important role in choriocapillaris dysfunction seen in SS-associated early AMD and implicate choroidal BM as a potential target for AMD management strategies.