Characterization of Potential Harm Caused by Electronic Cigarette Flavor Chemicals and their Reaction Products

项目摘要 电子烟(ECs)使用热气雾化通常含有高浓度 调味剂。该项目将购买650个商业电子液体：a)鉴定和量化的味道 所使用的化学品及其在蒸发过程中形成的降解反应产物(RXP)；以及b)评估 细胞对这些风味化学物质的反应通常为1毫克/毫升(即“主要风味化学品”，DFC)， 以及蒸发产生的气雾剂。将测试550种“流行的”加注和汽化器液体和100种液体。 据坊间报道，这些药物会导致使用者生病。工作将在两个校区进行： 波特兰州立大学的分析化学和加州大学河滨大学的人类毒物学。 目标1(PSU)。将对650种流体的DFC和蒸发RXP进行识别和量化。用来制造气雾剂 一系列不同功率的装置将被收集在异丙醇中，并用气相色谱进行分析。 质谱仪(GC/MS)和液相色谱/质谱仪(LLC/MS/MS)将按照标准进行鉴定。 由风味化学品形成的RXP将与EC溶剂形成的RXP明确区分开来。 目标2(UCR)。使用实验室制备的含有已确认的DFC的流体制成的VAPE气雾剂将由 一系列设备，并在使用体外试验的剂量反应实验中筛选细胞毒性 与正常人BEAS-2B肺上皮细胞结合。细胞毒性最强的气雾剂 将使用带有3D EpiAirway平台的气液界面曝光系统进行深入研究 重述人类呼吸道上皮细胞。单个DFC将使用六种与癌症相关的模式进行测试- 动作分析，包括我们观察到的上皮向间充质转化(EMT)的分析 一些EC产品。基因表达的改变将使用rna-seq和受影响的途径来确定。 已确认身份。DFC混合物将按照消费品中发现的相同相对比例进行测试。 目标3(UCR)。由于AIM 2将检查产生的热气溶胶，DFC及其RxPs产品都将 一直在现场。AIM 3将通过暴露3D EpiAirway平台来区分DFC和RXP毒性 不加热产生的气溶胶，含有加热产生的个别DFC或已确定的RXP。终结点 化验将按照目标2进行，以分离对个别化学品的毒性。 总结。对于ECS中使用的风味化学物质对健康的影响，人们知之甚少。这个项目将填补一个 与全面的新数据存在重要的信息差距。具体的毒物将被识别出来，其影响 将对混合DFC的数量进行评估。结果将发布在一个在线数据库上，任何人都可以访问 感兴趣的人或组织，并将为未来对风味的理解提供基础信息 烟草产品中的化学物质。这些数据是此类数据中的第一个，将有助于建立 美国国家卫生研究院可以制定未来的研究方向，以保护消费者的健康和保护接触者 通过二手和第三手手段。

Project Summary Electronic cigarettes (ECs) use heat to aerosolize “e-liquid” mixtures that usually contain high concentrations of flavor chemicals. This project will purchase 650 commercial e-liquids to: a) identify and quantify the flavor chemicals used plus their degradation reaction products (RxPs) formed upon vaping; and b) evaluate the cellular responses to those flavor chemicals often found at >1 mg/ml (viz. “dominant flavor chemicals”, DFCs), and to the aerosols produced on vaping. Tested will be 550 “popular” refill and cartomizer fluids and 100 fluids that have been anecdotally reported to cause sickness in users. Work will be conducted at two campuses: Portland State University for analytical chemistry, and University of California Riverside for human toxicology. Aim 1 (PSU). DFCs and vaping RxPs will be identified and quantified for the 650 fluids. Aerosols made using a range of devices at varied power will be collected in isopropanol and analyzed using gas chromatography/ mass spectrometry (GC/MS) and liquid chromatography/MS/MS. Identification will proceed with standards. RxPs formed from flavor chemicals will be unambiguously distinguished from RxPs from the EC solvents. Aim 2 (UCR). Vape aerosols made using lab-prepared fluids containing the identified DFCs will be made with a range of devices, and screened for cytotoxicity in dose-response experiments using in vitro assays in conjunction with BEAS-2B lung epithelial cells from normal human adults. The aerosols that are most cytotoxic will be studied in depth using an air-liquid interface exposure system with the 3D EpiAirway platform that recapitulates human respiratory epithelium. Individual DFCs will be tested using six cancer-related mode-of- action assays including assays for an epithelial to mesenchymal transition (EMT) that we have observed with some EC products. Alterations in gene expression will be determined using RNA-seq and affected pathways identified. Mixtures of DFCs will be tested at the same relative proportions found in consumer products. Aim 3 (UCR). Since Aim 2 will examine heat generated aerosols, both DFCs and their RxPs products will have been present. Aim 3 will differentiate the DFCs and RxP toxicities by exposing the 3D EpiAirway platform to aerosols generated without heat and containing individual DFCs or the identified RxPs from heating. Endpoint assays will proceed as in Aim 2 to isolate toxicity to individual chemicals. Summary. Little is known about the health effects of flavor chemicals used in ECs. This project will fill an important informational gap with comprehensive new data. Specific toxicants will be identified and the effects of mixing DFCs will be evaluated. Results will be posted on an online database that will be accessible to any interested persons or organization, and will provide foundational information for future understanding of flavor chemicals in tobacco products. The data, the first of their kind, will contribute to a foundation of science upon which the NIH can develop future research directions to protect consumers’ health and protect those exposed by second- and thirdhand means.