Investigating Impacts of Cannabis Tobacco and Vapes on Lung Cells Using GC×GC-MS

Importance of the Topic

Inhalation of tobacco smoke, electronic cigarette vapors and cannabis aerosols represents a major public health concern. Traditional cigarette smoking is linked to cardiovascular disease, lung cancer and chronic obstructive pulmonary disease, while the rapid rise of vaping and cannabis use has outpaced comprehensive toxicological evaluation. Understanding how complex aerosol mixtures alter lung cell metabolism is critical for risk assessment, regulatory decision-making and development of safer inhalation products.

Objectives and Study Overview

This study aimed to compare the biochemical impact of mainstream cigarette smoke, e-cigarette vapor and cannabis aerosol on two human lung cell models: bronchial epithelial cells (HBEC) and pulmonary neuroendocrine cells (ePNEC). Experiments addressed (1) time-dependent metabolic changes at 2 h and 6 h incubation, (2) differential response in healthy versus asthmatic cell lines, and (3) uniformity of smoke exposure across multiwell plates.

Methodology and Instrumentation

Cell cultures were exposed to controlled puffs generated by a programmable smoking machine equipped with a custom glass chamber and mass flow controllers. Smoke and vapor streams were maintained at 37 °C in a water-jacketed enclosure to mimic physiological conditions. Post-exposure, cell culture media were collected, metabolites were extracted, chemically derivatized and analyzed by comprehensive two-dimensional gas chromatography coupled to mass spectrometry (GC×GC-MS). Data processing pipelines deconvoluted more than 2 000 chromatographic features per sample and enabled comparative metabolomic profiling.

Main Findings and Discussion

• Time-dependent effects: Early (2 h) exposures induced mRNA-level changes, whereas extended (6 h) exposures produced pronounced alterations in metabolite profiles, particularly in ePNEC cells.
• Compound abundance patterns: Cigarette smoke elevated levels of tobacco alkaloids (nicotine), monoacetin and benzene derivatives, while vaping emissions were characterized by propylene glycol, ethanolamine and other humectant-derived species.
• Amino acid metabolism: Both cell types showed increased L-phenylalanine and tyrosine turnover, implicating altered phenylalanine hydroxylase activity and downstream catecholamine pathways (dopamine, norepinephrine).
• Health status comparison: Asthmatic HBEC and ePNEC cultures exhibited unique metabolic signatures compared to healthy cells, suggesting heightened susceptibility to aerosol-induced oxidative stress.
• Exposure uniformity: Custom 3D-printed enclosures and gradient testing confirmed even distribution of smoke across 24-well plates, minimizing well-to-well variability.

Benefits and Practical Applications

This GC×GC-MS approach provides high-resolution chemical fingerprints of inhalation exposures at the cellular level. It enables identification of hazardous constituents, mechanistic insights into metabolic pathways, and supports safety evaluation of emerging tobacco and cannabis products. The workflow is adaptable for QA/QC in product development, toxicology testing, and translational research.

Future Trends and Potential Uses

Integrating this platform with high-throughput 3D lung models and multiomic analyses (transcriptomics, proteomics) will deepen understanding of chronic exposure effects. Real-time aerosol monitoring and personalized cell models could advance risk assessment. Continued refinement of in vitro exposure systems and standardized analytical protocols will support regulatory frameworks and safer inhalation technologies.

Conclusion

Comprehensive GC×GC-MS profiling of lung cell exposures to cigarette smoke, e-cigarette vapor and cannabis aerosol reveals distinct metabolic perturbations influenced by exposure duration, cell type and health status. The methodology demonstrates robustness and scalability, warranting larger‐scale studies to inform public health and product regulation.

Instrumentation Used

• Customizable smoking/vaping machine with temperature-controlled glass chamber and mass flow controllers
• Circulating water bath (37 °C) and water-jacketed exposure enclosure
• Derivatization reagents for metabolite stabilization
• Comprehensive two-dimensional gas chromatograph (GC×GC) with mass spectrometer detector
• 3D-printed polycarbonate enclosure for exposure uniformity testing