Antibacterial Activity of Combination of Kaffir Lime Leaf (Citrus hystrix DC) and Cherry Leaf (Muntingia calabura L.) Extracts Against Staphylococcus aureus Bacteria
Abstract
The bacterial infection caused by Staphylococcus aureus can be treated with antibiotics. Kaffir lime (Citrus hystrix) and cherry (Muntingia calabura) leaves contain alkaloids, flavonoids, tannins, saponins, and terpenoids, known for their antibacterial properties. This study aimed to evaluate the antibacterial activity of a combination of kaffir lime and cherry leaf extracts against S. aureus. The kaffir lime leaves were extracted by maceration using 70% ethanol, while cherry leaves were extracted using 96% ethanol. The chemical compounds were identified, and antibacterial activity was tested by the dilution method to determine the Minimum Inhibitory Concentration (MIC). MIC values were combined in ratios of (1:1), (1:2), and (2:1). Antibacterial activity of the combinations was evaluated by the disk diffusion method. The most effective combination was further tested using the paper disc method for combination patterns. Data were analyzed statistically using the Shapiro-Wilk test, followed by one-way ANOVA and Post Hoc tests. The results showed the MIC for the kaffir lime leaf extract was 5%, and for cherry leaf extract, it was 10%. The disk diffusion test revealed the most effective combination was at a ratio of (2:1), with an inhibition zone of 19.35±0.33 mm, indicating a synergistic effect.
Keywords:
Staphylococcus aureus, Citrus hystrix DC, Muntingia calabura L., Extracts combination, Antibacterial activity
References
[1] Agustin, B. A., Puspawaty, N., Rukmana, R. M. (2018). Aktivitas Antibakteri Kombinasi Ekstrak Etanolik Daun Beluntas (Pluchaea indica Less.) dan Meniran (Phyllanthus niruri L.) terhadap Bakteri Staphylococcus aureus. Biomedika, 11(2), 79-87.
[2] Ngamsurach, P., & Praipipat, P. (2022). Antibacterial activities against Staphylococcus aureus and Escherichia coli of extracted Piper betle leaf materials by disc diffusion assay and batch experiments. RSC advances, 12(40), 26435-26454.
[3] Alouw, G., Fatimawali, F., Lebang, J. S. (2022). Uji Aktivitas Antibakteri Ekstrak Etanol Daun Kersen (Muntingia calabura L.) Terhadap Bakteri Staphylococcus aureus Dan Pseudomonas aeruginosa Dengan Metode Difusi Sumuran. Jurnal Farmasi Medica/Pharmacy Medical Journal (PMJ), 5(1), 36-44
[4] Kurnianingsih, D., Setiabudi, L., dan Tajudin, T. (2020). Uji efektivitas sediaan krim kombinasi ekstrak daun bakau hitam (Rhizophora mucronata) dan jeruk purut (Citrus hystrix) terhadap bakteri Staphylococcus aureus. Jurnal Ilmiah Jophus: Journal of Pharmacy UMUS, 2(01).
[5] Kandida, I., Tari, M., & Fatiqin, A. (2023). Effectiveness of the Combination of Green Betel Leaf Extract (Piper betle) and Mint Leaf (Mentha piperita) as Antibacterials against Streptococcus mutans. Bioactivities, 1(1), 32-38.
[6] Zhao, Z., Wang, Y., Nian M., Hong L., Chen, J., Qiao H,. Yang X., Li, X., Chen X., Zheng X., Wu X., (2023). Citrus hystrix: A review of phytochemistry, pharmacology and industrial applications research progress. Arabian Journal of Chemistry. 16
[7] Pohan, D. J., & Djojosaputro, M. (2021). Antibacterial Effectiveness of Extracts of Lime (Citrus aurantifolia Swingle) And Kaffir Lime (Citrus hystrix Dc) Leaves Against Escherichia coli. International Journal of Modern Pharmaceutical Research, 5(6), 29-36.
[8] Fitriyanti, F., Zannah, M., dan Nazarudin, M. (2024). Skrining Fitokimia Dan Efektivitas Antibakteri Ekstrak Etanol 70% Daun Jeruk Purut (Citrus hystrix DC) Terhadap Staphylococcus aureus. Sains Medisina, 2(3), 93-98.
[9] Bamasri, T. H. (2021). Daun Kersen (Muntingia calabura L.) sebagai antibakteri. Jurnal Penelitian Perawat Profesional, 3(2), 231-236.
[10] Gurning, K., Simanjuntak, H. A., Purba, H., Situmorang, R. F. R., Barus, L., & Silaban, S. (2021, March). Determination of total tannins and antibacterial activities ethanol extraction seri (Muntingia calabura L.) leaves. In Journal of Physics: Conference Series (Vol. 1811, No. 1, p. 012121). IOP Publishing.
[11] Binawati, D. K., dan Amilah, S. (2013). Effect of cherry leaf (Muntingia calabura) bioinsecticides extract towards mortality of worm soil (Agrotis ipsilon) and armyworm (Spodoptera exiqua) on plant leek (Allium fistolum). Wahana, 61(2).
[12] Nurjannah, I., Mustariani, B. A. A., dan Suryani, N. (2022). Skrining Fitokimia dan Uji Antibakteri Ekstrak Kombinasi Daun Jeruk Purut (Citrus hystrix) dan Kelor (Moringa oleifera L.) Sebagai Zat Aktif Pada Sabun Antibakteri. SPIN Jurnal kimia dan Pendidikan Kimia, 4(1), 23-36.
[13] Vakil, V., dan Trappe, W. (2019). Drug combinations: mathematical modeling and networking methods. Pharmaceutics, 11(5), 208.
[14] Friday, A., Edisha, G. D., Unekwuojo, A. B., Zainab, Y., dan Shadrach, A. U. (2021). Synergistic Efficacy Of Honey Single-Dose Combination Therapy: A Solution To Superbug Menaces. GSJ, 9(1).
[15] Nurafifah, D. A., Widyastuti, D. A., Minarti, I. B. (2021). Activity of Moringa oleifera seed ethanolic extract against E. coli. Advance Sustainable Science, Engineering and Technology. 3(2).
[16] Adlina, S., Vikandari, S. N., Fauziah, N. S., Endah, S. R. N. (2024). Potensi Ekstrak Daun Cherry (Mutingia calabura L.) sebagai Sumber Antioksidan Alami Dalam Sediaan Eye Gel. Jurnal Mandala Pharmacon Indonesia, 10(2), 715-721.
[17] Karlina, V. R., dan Nasution, H. M. (2022). Skrining Fitokimia Dan Uji Aktivitas Antibakteri Ekstrak Etanol Daun Jeruk Purut (Citrus hystrix DC) Terhadap Bakteri Staphylococcus aureus Dan Escherichia Coli. Journal of Health and Medical Science, 131-139.
[18] Maifitrianti, M., Sjahid, L. R., Nuroh, N., Acepa, R. A. M., & Murti, W. D. (2019). Aktifitas Antiinflamasi Fraksi-Fraksi Ekstrak Etanol 95% dari Daun Kersen (Muntingia Calabura L.) pada Tikus Putih Jantan. PHARMACY: Jurnal Farmasi Indonesia (Pharmaceutical Journal of Indonesia), 16(1), 1-16.
[19] Azhar, G., Ridwanto, R., Nasution, H. M., Pulungan, A. F., & Rani, Z. (2024). Antibacterial Activity of Kaffir Lime Leaf (Citrus hystrix DC) Ethanol Extract against Staphylococcus aureus and Escherichia coli. Indonesian Journal of Science and Pharmacy, 2(1), 16-21.
[20] Suryanita, S., Aliyah, A., Djabir, Y. Y., Wahyudin, E., Rahman, L., dan Yulianty, R. (2019). Identifikasi senyawa kimia dan uji aktivitas antioksidan ekstrak etanol kulit jeruk bali (Citrus maxima Merr.). Majalah Farmasi dan Farmakologi, 23(1), 16-20.
[21] Rai, A., & Khairnar, K. (2021). Isolation and characterization of bacteriophage against wastewater isolates of methicillin-resistant Staphylococcus aureus. Jundishapur Journal of Microbiology, 14(9).
[22] Triadi, B., Suwarno, S., Damayanti, R., Estoepangesti, A. T. S., Sugihartuti, R., & Sarudji, S. (2022). Antibiotic sensitivity test of Escherichia coli and Staphylococcus aureus isolated from the reproductive tract of dairy cows. Ovozoa: Journal of Animal Reproduction, 11, 72-80.
[23] Stain, G. (2024). Overview of Diagnostic Bacteriology and Mycobacteriology Methods Diagnostic Bacteriology Methods. Challenging Cases in Diagnostic Clinical Microbiology: Advanced Strategies and Techniques, 2.
[24] Astriani, N. K., Chusniasih, D., Marcellia, S. (2021). Uji Aktivitas Antibakteri Ekstrak Daun Jeruk Purut (Citrus hystrix DC) terhadap Bakteri Escherichia coli dan Staphylococcus aureus. Jurnal Ilmu Kedokteran dan Kesehatan, 8(3), 291-301.
[25] Pramiastuti, O., Rejeki, D. S., Maghfiroh, I., dan Firsty, G. R. (2020). Uji antibakteri kombinasi ekstrak daun belimbing wuluh (Averrhoa bilimbi L) dan daun kersen (Muntingia calabura L) Terhadap Staphylococcus aureus. Parapemikir: Jurnal Ilmiah Farmasi, 9(2), 33-41.
[26] Maimunah, S., Harefa, K., Yuliana, A., Ritonga, A. H., dan Hulu, A. (2019). Uji Aktivitas Antibakteri Ekstrak Daun Kersen (Muntingia calabura L.) Terhadap Bakteri Staphylococcus aureus Dan Methicillin-Resistant Staphylococcus aureus. Jurnal Farmanesia, 6(2), 101-108.
[27] Nourbakhsh, F., Lotfalizadeh, M., Badpeyma, M., Shakeri, A., & Soheili, V. (2022). From plants to antimicrobials: Natural products against bacterial membranes. Phytotherapy Research, 36(1), 33-52.
[2] Ngamsurach, P., & Praipipat, P. (2022). Antibacterial activities against Staphylococcus aureus and Escherichia coli of extracted Piper betle leaf materials by disc diffusion assay and batch experiments. RSC advances, 12(40), 26435-26454.
[3] Alouw, G., Fatimawali, F., Lebang, J. S. (2022). Uji Aktivitas Antibakteri Ekstrak Etanol Daun Kersen (Muntingia calabura L.) Terhadap Bakteri Staphylococcus aureus Dan Pseudomonas aeruginosa Dengan Metode Difusi Sumuran. Jurnal Farmasi Medica/Pharmacy Medical Journal (PMJ), 5(1), 36-44
[4] Kurnianingsih, D., Setiabudi, L., dan Tajudin, T. (2020). Uji efektivitas sediaan krim kombinasi ekstrak daun bakau hitam (Rhizophora mucronata) dan jeruk purut (Citrus hystrix) terhadap bakteri Staphylococcus aureus. Jurnal Ilmiah Jophus: Journal of Pharmacy UMUS, 2(01).
[5] Kandida, I., Tari, M., & Fatiqin, A. (2023). Effectiveness of the Combination of Green Betel Leaf Extract (Piper betle) and Mint Leaf (Mentha piperita) as Antibacterials against Streptococcus mutans. Bioactivities, 1(1), 32-38.
[6] Zhao, Z., Wang, Y., Nian M., Hong L., Chen, J., Qiao H,. Yang X., Li, X., Chen X., Zheng X., Wu X., (2023). Citrus hystrix: A review of phytochemistry, pharmacology and industrial applications research progress. Arabian Journal of Chemistry. 16
[7] Pohan, D. J., & Djojosaputro, M. (2021). Antibacterial Effectiveness of Extracts of Lime (Citrus aurantifolia Swingle) And Kaffir Lime (Citrus hystrix Dc) Leaves Against Escherichia coli. International Journal of Modern Pharmaceutical Research, 5(6), 29-36.
[8] Fitriyanti, F., Zannah, M., dan Nazarudin, M. (2024). Skrining Fitokimia Dan Efektivitas Antibakteri Ekstrak Etanol 70% Daun Jeruk Purut (Citrus hystrix DC) Terhadap Staphylococcus aureus. Sains Medisina, 2(3), 93-98.
[9] Bamasri, T. H. (2021). Daun Kersen (Muntingia calabura L.) sebagai antibakteri. Jurnal Penelitian Perawat Profesional, 3(2), 231-236.
[10] Gurning, K., Simanjuntak, H. A., Purba, H., Situmorang, R. F. R., Barus, L., & Silaban, S. (2021, March). Determination of total tannins and antibacterial activities ethanol extraction seri (Muntingia calabura L.) leaves. In Journal of Physics: Conference Series (Vol. 1811, No. 1, p. 012121). IOP Publishing.
[11] Binawati, D. K., dan Amilah, S. (2013). Effect of cherry leaf (Muntingia calabura) bioinsecticides extract towards mortality of worm soil (Agrotis ipsilon) and armyworm (Spodoptera exiqua) on plant leek (Allium fistolum). Wahana, 61(2).
[12] Nurjannah, I., Mustariani, B. A. A., dan Suryani, N. (2022). Skrining Fitokimia dan Uji Antibakteri Ekstrak Kombinasi Daun Jeruk Purut (Citrus hystrix) dan Kelor (Moringa oleifera L.) Sebagai Zat Aktif Pada Sabun Antibakteri. SPIN Jurnal kimia dan Pendidikan Kimia, 4(1), 23-36.
[13] Vakil, V., dan Trappe, W. (2019). Drug combinations: mathematical modeling and networking methods. Pharmaceutics, 11(5), 208.
[14] Friday, A., Edisha, G. D., Unekwuojo, A. B., Zainab, Y., dan Shadrach, A. U. (2021). Synergistic Efficacy Of Honey Single-Dose Combination Therapy: A Solution To Superbug Menaces. GSJ, 9(1).
[15] Nurafifah, D. A., Widyastuti, D. A., Minarti, I. B. (2021). Activity of Moringa oleifera seed ethanolic extract against E. coli. Advance Sustainable Science, Engineering and Technology. 3(2).
[16] Adlina, S., Vikandari, S. N., Fauziah, N. S., Endah, S. R. N. (2024). Potensi Ekstrak Daun Cherry (Mutingia calabura L.) sebagai Sumber Antioksidan Alami Dalam Sediaan Eye Gel. Jurnal Mandala Pharmacon Indonesia, 10(2), 715-721.
[17] Karlina, V. R., dan Nasution, H. M. (2022). Skrining Fitokimia Dan Uji Aktivitas Antibakteri Ekstrak Etanol Daun Jeruk Purut (Citrus hystrix DC) Terhadap Bakteri Staphylococcus aureus Dan Escherichia Coli. Journal of Health and Medical Science, 131-139.
[18] Maifitrianti, M., Sjahid, L. R., Nuroh, N., Acepa, R. A. M., & Murti, W. D. (2019). Aktifitas Antiinflamasi Fraksi-Fraksi Ekstrak Etanol 95% dari Daun Kersen (Muntingia Calabura L.) pada Tikus Putih Jantan. PHARMACY: Jurnal Farmasi Indonesia (Pharmaceutical Journal of Indonesia), 16(1), 1-16.
[19] Azhar, G., Ridwanto, R., Nasution, H. M., Pulungan, A. F., & Rani, Z. (2024). Antibacterial Activity of Kaffir Lime Leaf (Citrus hystrix DC) Ethanol Extract against Staphylococcus aureus and Escherichia coli. Indonesian Journal of Science and Pharmacy, 2(1), 16-21.
[20] Suryanita, S., Aliyah, A., Djabir, Y. Y., Wahyudin, E., Rahman, L., dan Yulianty, R. (2019). Identifikasi senyawa kimia dan uji aktivitas antioksidan ekstrak etanol kulit jeruk bali (Citrus maxima Merr.). Majalah Farmasi dan Farmakologi, 23(1), 16-20.
[21] Rai, A., & Khairnar, K. (2021). Isolation and characterization of bacteriophage against wastewater isolates of methicillin-resistant Staphylococcus aureus. Jundishapur Journal of Microbiology, 14(9).
[22] Triadi, B., Suwarno, S., Damayanti, R., Estoepangesti, A. T. S., Sugihartuti, R., & Sarudji, S. (2022). Antibiotic sensitivity test of Escherichia coli and Staphylococcus aureus isolated from the reproductive tract of dairy cows. Ovozoa: Journal of Animal Reproduction, 11, 72-80.
[23] Stain, G. (2024). Overview of Diagnostic Bacteriology and Mycobacteriology Methods Diagnostic Bacteriology Methods. Challenging Cases in Diagnostic Clinical Microbiology: Advanced Strategies and Techniques, 2.
[24] Astriani, N. K., Chusniasih, D., Marcellia, S. (2021). Uji Aktivitas Antibakteri Ekstrak Daun Jeruk Purut (Citrus hystrix DC) terhadap Bakteri Escherichia coli dan Staphylococcus aureus. Jurnal Ilmu Kedokteran dan Kesehatan, 8(3), 291-301.
[25] Pramiastuti, O., Rejeki, D. S., Maghfiroh, I., dan Firsty, G. R. (2020). Uji antibakteri kombinasi ekstrak daun belimbing wuluh (Averrhoa bilimbi L) dan daun kersen (Muntingia calabura L) Terhadap Staphylococcus aureus. Parapemikir: Jurnal Ilmiah Farmasi, 9(2), 33-41.
[26] Maimunah, S., Harefa, K., Yuliana, A., Ritonga, A. H., dan Hulu, A. (2019). Uji Aktivitas Antibakteri Ekstrak Daun Kersen (Muntingia calabura L.) Terhadap Bakteri Staphylococcus aureus Dan Methicillin-Resistant Staphylococcus aureus. Jurnal Farmanesia, 6(2), 101-108.
[27] Nourbakhsh, F., Lotfalizadeh, M., Badpeyma, M., Shakeri, A., & Soheili, V. (2022). From plants to antimicrobials: Natural products against bacterial membranes. Phytotherapy Research, 36(1), 33-52.
Published
2025-07-25
How to Cite
Az-Zahra, L. (2025). Antibacterial Activity of Combination of Kaffir Lime Leaf (Citrus hystrix DC) and Cherry Leaf (Muntingia calabura L.) Extracts Against Staphylococcus aureus Bacteria. Journal of Pharmaceutical Researchers, 3(2). https://doi.org/https://doi.org/10.31001/jpr.v2i2.2648
Section
Article
