Malays. J. Anal. Sci. Volume 29 Number 5 (2025): 1522

 

Research Article

 

Preliminary study on the rapid detection of eugenol in selected plants using thin layer chromatography

 

Wan Ahmad Kamil Wan Mustapha Albakri1, Nik Nor Izah Nik Ibrahim2 and Ruzilawati Abu Bakar2*

 

1Faculty of Science, Universiti Teknologi Malaysia, Skudai, Johor, Malaysia

2Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kelantan, Malaysia

 

*Corresponding author: ruzila@usm.my

 

Received: 24 March 2025; Revised: 19 August 2025; Accepted: 21 August 2025; Published: 16 October 2025

 

Abstract

Eugenol, a naturally occurring phenolic compound with significant pharmacological properties, is found in various aromatic plants. This study aims to develop and optimise a thin-layer chromatography (TLC) method for qualitative detection of eugenol in selected plant species. Samples were collected from Piper betle, Syzygium aromaticum, Cinnamomum zeylanicum, Zingiber officinale, Mitragyna speciosa, and Cymbopogon citratus. The plant extracts were prepared using a Soxhlet apparatus with 95% methanol as extraction solvent and subjected to TLC analysis. Silica gel plates were employed as the stationary phase, and a solvent system of dichloromethane: methanol (99:1) was optimised for the mobile phase. The detection of eugenol was carried out using ultraviolet (UV) light and fast blue BB spray for visualisation. The Rf values obtained were consistent with the standard eugenol reference, confirming the presence of eugenol in the selected plants. This TLC method offers a simple, cost-effective, and efficient approach for the rapid screening of eugenol in various plant materials.

 

Keywords: eugenol, plants, thin-layer chromatography, rapid detection

 

References

1.        Bisergaeva, R. A., Takaeva, M. A., and Sirieva, Y. N. (2021). Extraction of eugenol, a natural product, and the preparation of eugenol benzoate. Journal of Physics: Conference Series, 1889(2): 022085.

2.        Nisar, M. F., Khadim, M., Rafiq, M., Chen, J., Yang, Y., and Wan, C. C. (2021). Pharmacological properties and health benefits of eugenol: A comprehensive review. Oxidative Medicine and Cellular Longevity, 2021: 1-14.

3.        Jaganathan, S. K., and Supriyanto, E. (2012). Antiproliferative and molecular mechanism of eugenol-induced apoptosis in cancer cells. Molecules, 17(6): 6290-6304.

4.        Janssen, A. M., Scheffer, J. J. C., and Svendsen, A. B. (1987). Antimicrobial activities of essential oils. Pharmaceutical Weekblad, 9(4): 193-197.

5.        Kerosenewala, J., Vaidya, P., Ozarkar, V., Shirapure, Y., and More, A. P. (2023). Eugenol: Extraction, properties and its applications on incorporation with polymers and resins - A review. Polymer Bulletin, 80(14): 7047-7099.

6.        Elbestawy, M. K. M., El-Sherbiny, G. M., and Moghannem, S. A. (2023). Antibacterial, antibiofilm and anti-inflammatory activities of eugenol clove essential oil against resistant Helicobacter pylori. Molecules, 28(6): 2448.

7.        Marchese, A., Barbieri, R., Coppo, E., Orhan, I. E., Daglia, M., Nabavi, S. F., Izadi, M., Abdollahi, M., Nabavi, S. M., and Ajami, M. (2017). Antimicrobial activity of eugenol and essential oils containing eugenol: A mechanistic viewpoint. Critical Reviews in Microbiology, 43(6): 668-689.

8.        Devi, K. P., Nisha, S. A., Sakthivel, R., and Pandian, S. K. (2010). Eugenol (an essential oil of clove) acts as an antibacterial agent against Salmonella Typhi by disrupting the cellular membrane. Journal of Ethnopharmacology, 130(1): 107-115.

9.        Begum, S. N., Ray, A. S., and Rahaman, C. H. (2022). A comprehensive and systematic review on potential anticancer activities of eugenol: From pre-clinical evidence to molecular mechanisms of action. Phytomedicine, 107: 154456.

10.     National Center for Biotechnology Information. (2024). PubChem compound summary for CID 3314, Eugenol. PubChem. https://pubchem.ncbi.nlm.nih.gov/compound/Eugenol

11.     Aburel, O. M., Pavel, I. Z., Dănilă, M. D., Lelcu, T., Roi, A., Lighezan, R., Muntean, D. M., and Rusu, L. C. (2021). Pleiotropic effects of eugenol: The good, the bad, and the unknown. Oxidative Medicine and Cellular Longevity, 2021: 3165159.

12.     Catherine, A. A., Deepika, H., and Negi, P. S. (2012). Antibacterial activity of eugenol and peppermint oil in model food systems. Journal of Essential Oil Research, 24(5): 481-486.

13.     Balkrishna, A., Sharma, P., Joshi, M., Srivastava, J., and Varshney, A. (2021). Development and validation of a rapid high‐performance thin‐layer chromatographic method for quantification of gallic acid, cinnamic acid, piperine, eugenol, and glycyrrhizin in Divya‐Swasari‐Vati, an ayurvedic medicine for respiratory ailments. Journal of Separation Science, 44(17): 3146-3157.

14.     Murugananthan, Y., Razali, N. F. B., and Mansur, S. A. B. (2022). Comparison between eugenol in essential oil from cinnamon leaves (Cinnamomum verum) and clove (Eugenia caryophyllata) extracted by Soxhlet extraction. Progress in Engineering and Applied Technology, 3(1): 118-128.

15.     Kowalska, T., and Sajewicz, M. (2022). Thin-layer chromatography (TLC) in the screening of botanicals: Its versatile potential and selected applications. Molecules, 27(19): 6607.

16.     Sigma-Aldrich. Fast Blue BB salt hemi(zinc chloride) salt. Merck. https://www.sigma aldrich.com/MY/en/product/sigma/f3378

17.     Zollinger, H. (1952). The kinetics of the diazo coupling reaction. Chemical Reviews, 51(3): 347-361.

18.     dos Santos, N. A., Souza, L. M., Domingos, E., França, H. S., Lacerda, V., Beatriz, A., Vaz, B. G., Rodrigues, R. R. T., Carvalho, V. V., Merlo, B. B., Kuster, R. M., and Romão, W. (2016). Evaluating the selectivity of colorimetric test (Fast Blue BB salt) for the cannabinoids identification in marijuana street samples by UV–Vis, TLC, ESI(+)FT-ICR MS and ESI(+)MS/MS. Forensic Chemistry, 1: 13-21.

19.     Wiley. winCATS software. https://pro-4-pro.com/en/product/3761-camag-tlc-scanner-3-with-wincats-software-a-class-of-its-own.html

20.     Khalil, A. A., ur Rahman, U., Khan, M. R., Sahar, A., Mehmood, T., and Khan, M. (2017). Essential oil eugenol: Sources, extraction techniques and nutraceutical perspectives. RSC Advances, 7(52): 32669-32681.

21.     Zhou, L., Zheng, H., Tang, Y., Yu, W., and Gong, Q. (2013). Eugenol inhibits quorum sensing at sub-inhibitory concentrations. Biotechnology Letters, 35(4): 631-637.

22.     Cortés-Rojas, D. F., de Souza, C. R. F., & Oliveira, W. P. (2014). Clove (Syzygium aromaticum): A precious spice. Asian Pacific Journal of Tropical Biomedicine, 4(2): 90-96.

23.     Kramer, R. E. (1985). Antioxidants in clove. Journal of the American Oil Chemists’ Society, 62(1): 111-113.

24.     Fazal, F., Mane, P. P., Rai, M. P., Thilakchand, K. R., Bhat, H. P., Kamble, P. S., Palatty, P. L., and Baliga, M. S. (2014). The phytochemistry, traditional uses and pharmacology of Piper betel Linn. (betel leaf): A pan-asiatic medicinal plant. Chinese Journal of Integrative Medicine, 20(9): 707-720.

25.     Raja, M. C., Srinivasan, V., Selvaraj, S., and Mahapatra, S. (2015). Versatile and synergistic potential of eugenol: A review. Pharmaceutica Analytica Acta, 6(4): 1-6.

26.     Ulanowska, M., and Olas, B. (2021). Biological properties and prospects for the application of eugenol—A review. International Journal of Molecular Sciences, 22(7): 3671.

27.     Vangalapati, M., Satya, N. S., Prakash, D. S., & Avanigadda, S. S. (2012). A review on pharmacological activities and clinical effects of cinnamon species. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, 7(31): 1-11.

28.     Li, H., Liu, Y., Luo, D., Ma, Y., Zhang, J., Li, M., Yao, L., Shi, X., Liu, X., and Yang, K. (2019). Ginger for health care: An overview of systematic reviews. Complementary Therapies in Medicine, 45: 114-123.

29.     Adkins, J. E., Boyer, E. W., and McCurdy, C. R. (2011). Mitragyna speciosa, a psychoactive tree from Southeast Asia with opioid activity. Current Topics in Medicinal Chemistry, 11(9): 1165-1175.

30.     Shah, G., Shri, R., Panchal, V., Sharma, N., Singh, B., and Mann, A. S. (2011). Scientific basis for the therapeutic use of Cymbopogon citratus Stapf (lemon grass). Journal of Advanced Pharmaceutical Technology & Research, 2(1): 3-8.