*3.6. Data Treatment*

The data of the anti-aging activity of various enzymes were presented in three replications and subjected to one-way ANOVA followed by Duncan's test using CoStat version 6.311 (CoHort, Monterey, CA, USA, http://www.cohort.com).

A matrix of the concentration of a total of 30 major chemical compounds (>3%) identified in the EO of five *P. dioscoridis* ecospecies was constructed, these samples were (1) present sample; (2) purchased from a market in Cairo, Egypt; (3) collected from El-Sadat City, Egypt; (4) collected from Cairo-Suez desert road, Egypt dring April; and (5) collected from Turkey. While for *E. bonariensis*, a matrix of 27 major chemical compounds (>3%) represented six samples was designed, these samples were (1) present sample; (2) collected from Alexandria, Egypt; (3) collected from Monastir, Tunisia; (4) collected from Venezuela; (5) collected from Vietnam; and (6) collected from India. The matrices were subjected to principal component analysis (PCA) and agglomerative hierarchical clustering (AHC) via XLSTAT statistical computer software package (version 2018, Addinsoft Inc., New York, NY, USA).

## **4. Conclusions**

Herein, the GC-MS analysis of EOs of the above-ground parts of *P. dioscoridis* and *E. bonariensis*, revealed the identification of 29 and 35 compounds, respectively. Sesquiterpenes were characterized as the main components of EOs derived from the two plants. The major components of EO of *P. dioscoridis* were *α*-maaliene, berkheyaradulen, dehydrocyclolongifolene oxide, aromadendrene oxide-2, and *β*-muurolene. While, *trans*-α-farnesene, *O*-ocimene, and α-maaliene represented the abundant constituents of *E. bonariensis* EO. The observed variation in the EOs composition among the studied ecospecies and that reported support the changing of the taxa names. EO of *P. dioscoridis* exhibited cytotoxicity against the two cancer cells, MCF-7 and A-549, while the EO of the *E. bonariensis* showed activity only against HepG2. The EOs of *P. dioscoridis* and *E. bonariensis* as well as the mixture of them (1:1), exhibited significant anti-aging activity in which the mixture (1:1) > *P. dioscoridis* > *E. bonariensis*. All these data deduced the studied EOs of these two plants may be used as antiaging and anticancer leading agents.

**Author Contributions:** Conceptualization, A.M.A.-E., A.M.E. and A.I.E.; methodology, A.M.E., R.F.A., A.M.A.-E., A.E.-N.G.E.-G., A.I.E., and M.I.N.; data curation, A.M.A.-E. A.I.E.; software, A.M.A.-E., A.E.-N.G.E.-G. and A.M.E.; validation, A.M.E., R.F.A., A.M.A.-E., A.E.-N.G.E.-G., A.I.E. and M.I.N.; formal analysis, A.M.E., R.F.A., A.M.A.-E., A.E.-N.G.E.G., A.I.E. and M.I.N.; investigation, A.M.E., R.F.A., A.M.A.-E., A.E.-N.G.E.-G., A.I.E. and M.I.N.; funding acquisition, A.M.E.; writing—original draft preparation, A.M.A.-E. and A.I.E.; writing—review and editing, A.M.A.-E., R.F.A., A.M.A.-E., A.E.-N.G.E.-G., M.I.N. and A.I.E. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research received no external funding.

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** The data presented in this study are available in the article.

**Acknowledgments:** The authors gratefully acknowledge the National Research Centre, Cairo, Egypt for the support.

**Conflicts of Interest:** The authors declare no conflict of interest.

**Sample Availability:** Samples of the compounds are not available from the authors.
