LC–MS and GC–MS Reveal Enhanced Anti-Inflammatory Metabolites in Callus Cultures of Eucalyptus camaldulensis

Researchers evaluating leaf and tissue culture (callus) extracts and corresponding volatile oils from Eucalyptus camaldulensis Dehn (Family Myrtaceae), extensively exploited for its anti-inflammatory efficacy, subjected the methanolic extracts from both plant sources to liquid chromatography–mass spectrometry (LC–MS). Their findings underscore the potential of in vitro tissue culture as a biotechnological tool for amplifying the yield and bioactivity of secondary metabolites in E. camaldulensis. A paper based on their research was published in Scientific Reports. (1)

E. camaldulensis has traditionally been used as an anti-inflammatory, antispasmodic, antipyretic, and antimalarial remedy,as well as for respiratory infectious diseases such as pulmonary tuberculosis; in addition, E. camaldulensis helps decrease various clinical respiratory symptoms such as nasal sinus infections and coughs. (2,3)

The research team’s analysis revealed 32 phytoconstituents in leaf extracts and 54 in callus extracts, with identified chemical classes including flavonoids, tannins, coumarins, and phenolic acids, with higher relative abundance in callus-derived extracts. Volatile oil analysis using gas chromatography–mass spectrometry (GC–MS) identified 58 compounds in leaf oil and 52 in callus oil. Quantitative profiling demonstrated a significant elevation in bioactive volatiles within callus oil: 1,8-cineole content showed a 2.1-fold increase compared to leaf oil, and both α-terpineol and sabinene exhibited approximately fourfold increases relative to leaf oil. Bioassays further indicated that callus extracts and volatile oils possess enhanced anti-inflammatory and antioxidant activities, exhibiting higher potency than leaf-derived counterparts. Remarkably, the anti-inflammatory activity of callus volatile oil exceeded that of the standard pharmaceutical agent indomethacin. (1)

The authors of the study write that their efforts “demonstrated the first comprehensive phytochemical investigation into the enhancement of secondary metabolite production of Eucalyptus camaldulensis using suspension culture technique. The results demonstrate that callus extracts contain significantly higher concentrations of key bioactive compounds, such as 1,8-cineole, α-terpineol, and sabinene, compared to leaf extracts. Bioassay analyses revealed that callus-derived volatile oil exhibited superior anti-inflammatory and antioxidant activities, with its anti-inflammatory effect surpassing that of piroxicam. These findings underscore the potential of in vitro tissue culture as an effective biotechnological approach to enhancing the yield and bioactivity of secondary metabolites in E. camaldulensis, paving the way for the development of more potent natural therapeutic agents.” (1)

The researchers believe that additional in vivo investigations are warranted to validate the therapeutic efficacy and safety of the bioactive extracts. In addition, complementary advanced statistical analyses are recommended for the identification and prioritization of the lead compounds contributing to the observed biological activities. They recommend that future studies focus on the optimization of culture conditions through the strategic use of various plant growth regulators, with the aim being to improve both the yield and specificity of targeted phytochemicals. Finally, mechanistic studies at the cellular and molecular levels, in their opinion, are essential to clarify the underlying pathways mediating the biological effects of these extracts. (1)

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References

1. Mahrous, M. H.; Nassar, A. M.; El-Fiky, F. K. et al. Enhancing Anti-Inflammatory Activity of Eucalyptus camaldulensis by Upregulating Secondary Metabolites Using Suspension Cultures Techniques. Sci Rep. 2026.DOI: 10.1038/s41598-025-34963-8
2. Kheder, D. A.; Al-Habib, O. A. M.; Gilardoni, G. et al. Components of Volatile Fractions from Eucalyptus camaldulensis Leaves from Iraqi-Kurdistan and Their Potent Spasmolytic Effects. Molecules 2020, 25 (4), 804. DOI: 10.3390/molecules25040804
3. Dheyab, A. S.; Ibrahim A. J. K.; Aljumily, E. K. et al. Antimycobacterial Activity and Phytochemical Properties of Eucalyptus camaldulensis (Eucalyptus) Extracted by Deep Eutectic Solvents. Materials Today: Proceedings 2022, 65, 2738–2742. DOI: 10.1016/j.matpr.2022.06.017