Antibacterial Activity, TLC-Bioautography Analysis, and Determination of Bioactive Components in Ethyl Acetate Extract of Robusta Coffee Leaf (Coffea canephora L.) From Aceh, Indonesia


  • Rasidah Rasidah Department of Pharmacy, Aceh Health Polytechnic, Aceh Besar, Aceh, Indonesia.
  • Suryawati Suryawati Faculty of Medicine, Syiah Kuala University, Aceh, Indonesia.
  • Munira Munira Department of Pharmacy, Aceh Health Polytechnic, Aceh Besar, Aceh, Indonesia.
  • Noni Zakiah Department of Pharmacy, Aceh Health Polytechnic, Aceh Besar, Aceh, Indonesia.
  • Sri Rahayu Department of Pharmacy, Aceh Health Polytechnic, Aceh Besar, Aceh, Indonesia.
  • Vonna Aulianshah Department of Pharmacy, Aceh Health Polytechnic, Aceh Besar, Aceh, Indonesia.


Spectrophotometry, Total flavonoids, TLC-Bioautography, Coffea canephora L, Antibacterial


Infectious diseases affect many people in the world, including Indonesia. One of the causes of infection is bacteria, including scherichia coli and Staphylococcus aureus. Robusta coffee (Coffea canephora L.) is one of the plants that is widely encountered on Indonesian plantations. The leaves are usually discarded after being trimmed to maximize the growth of coffee beans. The results of several studies show that robusta coffee leaves demonstrate antibacterial and antioxidant activity due to the presence of flavonoids, alkaloids, tannins, saponins, polyphenols, and chlorogenic acids. The aim of this research to investigate antibacterial activities, total flavonoid content and inspect the phytochemicals of the bioactive compounds as antibacterial with TLC Bioautography in the ethyl acetate extract of robusta coffee leaves. The identification of secondary metabolites was conducted by using the TLC-Bioautography method against Escherichia coli and Staphylococcus aureus. The determination of total flavonoid levels was carried out by using Spectrophotometry UV-Vis. Phytochemical testing showed that the extract contained alkaloids, steroids, phenolics, tannins, and flavonoids. The ethyl acetate extract inhibits the growth of Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 (P=0.00) with 100% extract as the most potent. The results of TLC-Bioautography showed the presence of polyphenolic compounds (Rf 0.36) which was confirmed by a clear zone in the disc diffusion test. The total flavonoid content in robusta coffee leaves was 57.255±0.0348 mg QE/g extract. Ethyl acetate extract from coffee leaves (Coffea canephora L). inhibits the growth of S. aureus and E. coli due to the presence of polyphenolic compounds


Muslim Z, Dephinto Y. Antibacterial Activity Of Robusta Coffee (Coffea canephora L.) Leaves To Staphylococcus aureus And Escherichia coli. Asian J. Pharm. Clin. Res. 2019; 12(12):113-115.

Anjani G, Widyastuti N, Masruroh Z, Yuliana RAD, Almira VG, Tsani AFA, Nissa C, Prawira-Atmaja MI. Bioactive components and antibacterial activity in Robusta coffee leaves (Coffea canephora). Int J Pharm Res. 2020;12(3):1374-1382.

Portillo OR, Arévalo AC. Coffee’s Phenolic Compounds. A general overview of the coffee fruit’s phenolic composition. Bionatura. 2022;7(3):1-19.

Limón-Rivera R, Mejía-Ochoa FJ, Peláez-Muñoz EC, ArgasRivera Y, Hernández-Velásquez S, Fierro-Domínguez E, Castro-Medina L, González-Pérez M. Analysis of the Relationship between Lifestyle and Coffee Consumption Habits, from the Myth Approach, in the Municipalities of

Orizaba, Tehuipango, and Zongolica Veracruz. Int J Adv Eng. 2017;3(12):1111-1121.

Rosyidi D, Radiati LE, Qosimah D, Amri IA, Prasetyo D, Anisa AK. The Potential of Lampung Robusta Green Coffee (Coffea canephora) Extract Toward T Cell Activation in ISA-brown Laying Chickens. Trop J of Nat Prod Res. 2022;6(6):875-878

Maxiselly Y, Anusornwanit P, Rugkong A, Chiarawipa R, Chanjula P. Morpho-Physiological Traits, Phytochemical Composition, and Antioxidant Activity of Canephora Coffee Leaves at Various Stages. Int J Plant Biol. 2022;13(2):106-114.

Farhadi F, Khameneh B, Iranshahi M, Iranshahy M. Antibacterial activity of flavonoids and their structure–activity relationship: An update review. Phytotherapy Research. 2018;1(1):1-28.

Do QD, Angkawijaya AE, Tran-Ngunyen PL, Huynh LH, Soetaredjo FE, Ismadji S, Ju YS. Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. J Food Drug Anal. 2014;22(3):296-302.

Kemenkes. Farmakope Herbal Indonesia Edisi II. 2017; 531.

Shaikh JR, Patil MK. Qualitative tests for preliminary phytochemical screening: An overview. Int J Chem Stud. 2020;8(2):603-608.

Syafriana V, Purba RN, Djuhariah YS. Antibacterial Activity of Kecombrang Flower ( Etlingera elatior (Jack) R.M. Sm) Extract against Staphylococcus epidermidis and Propionibacterium acnes. J Trop Biodivers Biotechnol. 2021;6(1):1-11.

Wang M, Zhang Y, Wang R, Wang R, Yang B, Kuang H. An evolving technology that integrates classical methods with continuous technological developments: Thin-layer chromatography bioautography. Molecules. 2021;26(July):4647-4668.

Choma IM and Grzelak EM. Bioautography detection in thin-layer chromatography. J Chromatogr A. 2011;1218(1):2684-2691.

Dewanjee S, Gangopadhyay M, Bhattacharya N, Khanra R, Dua TK. Bioautography and its scope in the field of natural product chemistry. J Pharm Anal. 2015;5(2):75-84.

Jesionek W, Grzelak EM, Majer-Dziedzi B, Chom IM. Thinlayer chromatography-Direct bioautography for the screening of antimicrobial properties of plant extracts. J Planar Chromatogr-Mod TLC. 2013;26(2):109-113.

Setyati D, Sulistiyowati H, Erizcy MP, Ratnasari T. The Flavonoid and Alkaloid Content Of Cyclosorus parasiticus(Linn.) Farwell Ferns at The Plantation Areas Of Jember Regency. Biolink. 2020;7(1):23–37.

Gopinath B, McMahon C, Lewis J, Bondonno N, Bondonno C, Burlutsky G, Hodgson J, Mitchell P. Associations between intake of dietary flavonoids and 10-year incidence of age-related hearing loss. Nutrients. 2020;12(11):1-8.

Abubakar AR, Haque M. Preparation of medicinal plants: Basic extraction and fractionation procedures for experimental purposes. J Pharm and Bioallied Sci. 2020;12(1):1-10.

Dias MC, Pinto DCGA, Silva AMS. Plant flavonoids: Chemical characteristics and biological activity. Molecules. 2021;26(17):1-16.

Jubeh B, Breijyeh Z, Karaman R. Resistance of grampositive bacteria to current antibacterial agents and overcoming approaches. Molecules. 2020;25(12):2888-2910.

Paputungan WA, Lolo WA, Siampa JP. Antibacterial Activity and TLC-Bioautographic Analysis of the Fraction of Robusta Coffee Beans (Coffea canephora Pierre ex A. Froehner). Pharmacon. 2019;8(3):516–524.

Fatmawati S, Ermi HNP, Fadillah A, Putri AM. Antioxidant Activity and Sun Protection Factor (SPF) Graded Extract of Katuk Leaves (Sauropus androgynus (L.) Merr.). IOP Conf Ser Earth Environ Sci. 2022;1041(1):1-9.

Croce AC. Light and Autofluorescence, Multitasking Features in Living Organisms. Photochem. 2021;1(2):67-124.

Aminah A, Tomayahu N, Abidin Z. Determination of Total Flavonoid Content of Ethanol Extract of Avocado Fruit Peel (Persea americana Mill.) Using UV-VIS Spectrophotometry Method. J Fitofarmaka Indones. 2017;4(2):226-230.

Flemming J, Meyer-Probst CT, Speer K, Kölling-Speer I, Hannig C, Hannig M. Preventive applications of polyphenols in dentistry-a review. Int J Mol Sci. 2021;22(May):4892-4923.

Nur S, Mubarak F, Jannah C, Winarni D, Rahman DA, Hamdayani LA, Sami FJ. Total phenolic and flavonoid compounds, antioxidant and toxicity profile of extract and fractions of paku atai tuber (Angiopteris ferox Copel). Food Res. 2019;3(6):734-740.

Maungchanburee S, Phongseeput S, Thongsri O, Maijuy M, Chaithada P. Study of antioxidant activities, total phenolic content and total flavonoid content of the extracts of Monochoria vaginalis and Cissus repens Lamk. using different solvents. J. Pharm. Sci & Res. 2020;12(3):356-359.

Doloking H, Mukhriani M, Ningsi S, Tahar N. Flavonoids: A Review on Extraction, Identification, Quantification, and Antioxidant Activity. Ad-Dawaa’ J.Pharm. Sci.2022;5(1):38-63.



How to Cite

Rasidah, R., Suryawati, S., Munira, M., Zakiah, N., Rahayu, S., & Aulianshah, V. (2023). Antibacterial Activity, TLC-Bioautography Analysis, and Determination of Bioactive Components in Ethyl Acetate Extract of Robusta Coffee Leaf (Coffea canephora L.) From Aceh, Indonesia: Tropical Journal of Natural Product Research (TJNPR), 7(4), 2760–2764. Retrieved from