Chromatography-Based Metabolomics Reveals Dynamic Aroma Changes During Cigar Aging

A joint study by the China National Tobacco Corporation (Zhengzhou) and the Sichuan China Tobacco Industry Company, (Chengdu) used an untargeted metabolomics method based on headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) to comprehensively profile volatile compounds and track dynamic changes during cigar aging, which plays a critical role in enhancing flavor complexity and overall quality of cigars. A paper detailing this research was published in Frontiers in Plant Science (1).

Cigars are widely valued for distinctive flavor and aroma (2,3). The production process involves several stages, including cultivation, curing, fermentation, rolling, and aging of tobacco (4,5). Recently, advancements in analytical methods have been employed in the exploration of chemical transformations and mechanisms pertaining to cigar aging. Zhu and colleagues used headspace solid-phase microextraction coupled with gas chromatography-ion mobility spectrometry (HS-GC-IMS) in the analysis of volatiles under varying aging conditions, identifying aging time as the most influential factor in aroma differentiation (6). Hu and colleagues utilized sensory analysis, chemical profiling, and microbial sequencing to gauge the impact of different aging media and found that a “coffee” media appreciably heightened the complexity of the aroma as well as altered the surface microbiota within 30 days (7). Xue and colleagues exposed how aging environments regulate microbial successions and the degradation of sugars and alkaloids, emphasizing the synergistic effect of microbials and chemical transformations in the development of cigar flavor development (8). Despite these instances, current research mainly focuses on short-term aging (<90 days) or specific aging conditions, with limited insights into the long-term dynamics of aroma compounds, the identification of key volatiles, and their functional roles during extended aging, thus inspiring this joint study (1).

A total of 1836 volatile compounds were identified using the method developed by the team, primarily comprising heterocyclics, terpenoids, ketones, and esters. Multivariate analyses, including principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA), segmented the aging process into four distinct stages, with most differential metabolites showing upregulated trends. Notably, terpenoids exhibited substantial increases in both diversity and abundance. KEGG pathway enrichment analysis highlighted significant involvement of sesquiterpenoid and triterpenoid biosynthesis pathways in cigar aging process. Analysis based on relative odor activity values (rOAV) indicated a progressive enhancement of fruity, floral, honey, woody, and sweet notes, while coffee, roasted, hay, burnt, and spicy aromas declined over time. By integrating rOAV data with K-means clustering analysis, 21 key aroma-active compounds were identified to be closely associated with the aroma changes during aging, including 14 consistently upregulated compounds, such as (E)-β-damascone and δ-cadinene, and 7 downregulated ones, for example, 2-ethyl-3,5-dimethylpyrazine, 3-octen-2-one (1).

These findings provide new insights into the metabolic basis of cigar aging. The team recommends that future studies focus on the regulatory mechanisms of key enzymes and the biosynthesis of FPP in the sesquiterpenoid pathway, aiming to achieve precise modulation of cigar aroma and process optimization (1).

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References

• Yang, M.; Zhou, H.; Hu, X. et al. Untargeted Metabolomics Based on HS- SPME-GC-MS Revealing the Dynamic Evolution of Aroma Components During Cigar Aging. Front. Plant Sci. 2025, 16, 1657415. DOI: 10.3389/fpls.2025.1657415
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• Yang, L.; Liu, L.; Ji, L. et al. Analysis of Differences in Aroma and Sensory Characteristics of the Mainstream Smoke of Six Cigars. Heliyon 2024, 10 (4), e26630. DOI: 10.1016/j.heliyon.2024.e26630
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• Zhu, B.; Chen, J.; Song, G. et al. Characterization of Volatile Flavor Compounds of Cigar with Different Aging Conditions by Headspace–Gas Chromatography–Ion Mobility Spectrometry. Open Chem. 2025, 23, 20240129. DOI: 10.1515/chem-2024-0129

Hu, W.; Cai, W.; Li, D. et al. Exogenous Additives Facilitate the Fermentation of Cigar Tobacco Leaves: Improving Sensory Quality and Contents of Aroma Components. Food Sci. Technol. 2022a, 42, e68122. DOI: 10.1590/fst.68122

Xue, F.; Yang, J.; Luo, C. et al Metagenomic Insight into the Biodegradation of Biomass and Alkaloids in the Aging Process of Cigar. Bioresour. Bioprocess. 2023, 10 (1), 45. DOI: 10.1186/s40643-023-00667-y