Antibacterial Synergy Detection of Lignin Extract from Oil Palm Empty Fruit Bunches (Opefb) Combined with Amoxicillin Against Staphylococcus Aureus Using The Azdast Method


Rifki Anshory Hendri
Herman Suryadi
Hayun Hayun


Oil palm empty fruit bunches (OPEFB) are waste generated by the palm oil industry and are generally considered an environmental problem due to their large quantity and difficult recyclability. This study's objective was to examine the potential of OPEFB lignin extract combined with amoxicillin in inhibiting the growth of Staphylococcus aureus bacteria. The Synergism Test of the OPEFB lignin extract-antibiotic combination was conducted using the AZDAST method (Ameri Ziaei Double Antibiotic Synergism Test). The lignin extraction process was a steam process using a 4% oxalic acid solution under optimal conditions. Then, Calcium ions precipitate 4% oxalate by adding Ca(OH)2 to the extract. The Lignin content was determined by UV-Vis spectrophotometry at 280 nm, and then the lignin extract was combined with antibiotics for antibacterial assay. The results showed a lignin content of 1.06 %. The AZDAST test results revealed clear zones from the combination of OPEFB lignin extract and amoxicillin with a diameter of 20.23 mm for the extract containing oxalate and 15.83 mm for the oxalate-free extract. Meanwhile, on average, the clear zone of single amoxicillin was only 14.13 mm. Based on these results, the combination of OPEFB lignin extract with amoxicillin was stronger than amoxicillin alone in inhibiting the growth of S. aureus, but it was not significantly different (p-value> 0.05). In conclusion, the potential of OPEFB lignin extract synergistically worked with the antibiotic amoxicillin in inhibiting the growth of S. aureus.


Author Biographies

Rifki Anshory Hendri, Universitas Indonesia

Faculty of Pharmacy

Herman Suryadi, Universitas Indonesia

Laboratory of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmacy

Hayun Hayun, Universitas Indonesia

Laboratory of Pharmaceutical-Medicinal Chemistry and Bioanalysis, Faculty of Pharmacy

How to Cite
Hendri, R. A., Suryadi, H., & Hayun, H. (2024). Antibacterial Synergy Detection of Lignin Extract from Oil Palm Empty Fruit Bunches (Opefb) Combined with Amoxicillin Against Staphylococcus Aureus Using The Azdast Method. Journal of Applied Agricultural Science and Technology, 8(1), 117-129.


  1. Alzagameem, A., Klein, S. E., Bergs, M., Do, X. T., Korte, I., Dohlen, S., Hüwe, C., Kreyenschmidt, J., Kamm, B., Larkins, M., & Schulze, M. (2019). Antimicrobial activity of lignin and lignin-derived cellulose and chitosan composites against selected pathogenic and spoilage microorganisms. Polymers, 11(4), 670.
  2. BPS. (2022). Statistik Kelapa Sawit Indonesia 2021. Jakarta, Indonesia: BPS – Statistics Indonesia.
  3. Bukhari, N. A., Loh, S. K., Luthfi, A. A. I., Nasrin, A. B., Abdul, P. M., Harun, S., & Jahim, J. M. (2021). Oil palm trunk biomass pretreatment with oxalic acid and its effect on enzymatic digestibility and fermentability. Materials Today: Proceedings, 42, 119–123.
  4. Dirgantoro, M. A., & Adawiyah, R. (2018). Nilai Ekonomi Pemanfaatan Limbah Kelapa Sawit Menuju Zero Waste Production. Biowallacea, 5(2), 825–837.
  5. Dong, X., Dong, M., Lu, Y., Turley, A., Jin, T., & Wu, C. (2011). Antimicrobial and antioxidant activities of lignin from residue of corn stover to ethanol production. Industrial Crops and Products, 34(3), 1629–1634.
  6. Hidayah, N., & Wusko, I. U. (2020). Characterization and Analysis of Oil Palm Empty Fruit Bunch (OPEFB) Waste of PT Kharisma Alam Persada South Borneo. Majalah Obat Tradisional, 25(3).
  7. Kai, D., Chow, L. P., & Loh, X. J. (2018). Lignin and Its Properties. Retrieved from
  8. Kalogiannis, K. G., Matsakas, L., Aspden, J., Lappas, A. A., Rova, U., & Christakopoulos, P. (2018). Acid assisted organosolv delignification of beechwood and pulp conversion towards high concentrated cellulosic ethanol via high gravity enzymatic hydrolysis and fermentation. Molecules, 23(7), 1647.
  9. Lee, R. A., Bédard, C., Berberi, V., Beauchet, R., & Lavoie, J. M. (2013). UV-Vis as quantification tool for solubilized lignin following a single-shot steam process. Bioresource Technology, 144, 658–663.
  10. Lindsay, A. C., Kudo, S., & Sperry, J. (2019). Cleavage of lignin model compounds and ligninox using aqueous oxalic acid. Organic and Biomolecular Chemistry, 17(31), 7408–7415.
  11. Lu, F., Wang, C., Chen, M., Yue, F., & Ralph, J. (2021). A facile spectroscopic method for measuring lignin content in lignocellulosic biomass. Green Chemistry, 23(14), 5106–5112.
  12. Morena, A. G., & Tzanov, T. (2022). Antibacterial lignin-based nanoparticles and their use in composite materials. Nanoscale Advances, 4(21), 4447–4469).
  13. Risanto, L., Hermiati, E., & Sudiyani, Y. (2014). Properties of Lignin from Oil Palm Empty Fruit Bunch and Its Application for Plywood Adhesive. Makara Journal of Technology, 18(2).
  14. Ruwoldt, J., Tanase-Opedal, M., & Syverud, K. (2022). Ultraviolet Spectrophotometry of Lignin Revisited: Exploring Solvents with Low Harmfulness, Lignin Purity, Hansen Solubility Parameter, and Determination of Phenolic Hydroxyl Groups. ACS Omega, 7(50), 46371–46383.
  15. Saquib, S. A., AlQahtani, N. A., Ahmad, I., Arora, S., Asif, S. M., Javali, M. A., & Nisar, N. (2021). Synergistic antibacterial activity of herbal extracts with antibiotics on bacteria responsible for periodontitis. Journal of Infection in Developing Countries, 15(11), 1685–1693.
  16. Suryadi, H., Yanuar, A., Harmita, & Rachmadani, P. W. (2020). Response Surface Methodology Applied to Oxalic Acid Hydrolysis of Oil Palm Empty Fruit Bunch Biomass for D-Xylose Production. International Journal of Applied Pharmaceutics, 12(1), 172–176.
  17. Tribot, A., Amer, G., Alio, M. A., de Baynast, H., Delattre, C., Pons, A., Mathias, J. D., Callois, J. M., Vial, C., Michaud, P., & Dussap, C. G. (2019). Wood-lignin: Supply, extraction processes and use as bio-based material. In European Polymer Journal, 112, 228–240.
  18. Wang, G., Pang, T., Xia, Y., Liu, X., Li, S., Parvez, A. M., Kong, F., & Si, C. (2019). Subdivision of bamboo kraft lignin by one-step ethanol fractionation to enhance its water-solubility and antibacterial performance. International Journal of Biological Macromolecules, 133, 156–164.
  19. Wei, X., Cui, S., & Xie, Y. (2022). Synthesis and Antibacterial Properties of Oligomeric Dehydrogenation Polymer from Lignin Precursors. Molecules, 27(5), 1466.
  20. Yun, J., Wei, L., Li, W., Gong, D., Qin, H., Feng, X., Li, G., Ling, Z., Wang, P., & Yin, B. (2021). Isolating High Antimicrobial Ability Lignin from Bamboo Kraft Lignin by Organosolv Fractionation. Frontiers in Bioengineering and Biotechnology, 9.
  21. Ziaei-Darounkalaei, N., Ameri, M., Zahraei-Salehi, T., Ziaei-Darounkalaei, O., Mohajer-Tabrizi, T., & Bornaei, L. (2016). AZDAST the new horizon in antimicrobial synergism detection. MethodsX, 3, 43–52.