Inhibitory activity of several extract of Piper betle Leaf against S. aureus

Authors

  • Ahmad Shobrun Jamil Departmet of Pharmaceutical Biology Faculty of Health Science, University of Muhammadiyah Malang
  • Siti Rofida Departmet of Pharmaceutical Biology Faculty of Health Science, University of Muhammadiyah Malang
  • Dinda Farida Departmet of Pharmaceutical Biology Faculty of Health Science, University of Muhammadiyah Malang
  • Dwi Retno Nur Syahida Department of Pharmaceutical Chemistry Faculty of Health Science, University of Muhammadiyah Malang
  • Trimianti Hidahyatun Nazah Department of Pharmaceutical Chemistry Faculty of Health Science, University of Muhammadiyah Malang

DOI:

https://doi.org/10.12928/pharmaciana.v11i2.16999

Keywords:

antimicrobial, ethyl acetate, n-hexane, Piper betle, S. aureus

Abstract

A high number of infections from year to year require infectious diseases to get serious attention. The antimicrobial compound exploration must be continued to anticipate the development of infectious diseases. The purpose of this study was to find out in vitro antimicrobial activity of n-hexane, ethyl acetate, and ethanol fractions of Piper betle green leaves against S. aureus, and profiles of secondary metabolite compounds contained in these three extracts. The antimicrobial activity test was carried out by disk diffusion test of the fraction of n-hexane, ethyl acetate, and ethanol at a concentration of 6,25, 12,5, and 18,75 µg/disk. Detection of secondary metabolite content was done by the Thin Layer Chromatography method with stain-view reagents. The highest antimicrobial activity was found in the ethyl acetate fraction and was significantly different from the activity in the ethanol and n-hexane fractions. However, the antibacterial activity of all fractions was lower compared to the positive control of amoxiclav 30µg/disk. The phytochemical screening results of secondary metabolites of each fraction were shown that the n-hexane fraction contained alkaloids, terpenoids, flavonoids, polyphenols, and anthraquinone; the ethyl acetate fraction contained terpenoids, polyphenols, and anthraquinone; while the ethanol fraction contained alkaloids, terpenoids, polyphenols, and anthraquinone. Based on the test results, it is concluded that all fractions of Piper betle leaf extract had high antimicrobial activity; meanwhile, the ethyl acetate fraction had the highest activities among others. Each fraction was proven to have a different composition of secondary metabolites.

References

Ajileye, O. O., Obuotor, E. M., Akinkunmi, E. O., & Aderogba, M. A. (2015). Isolation and characterization of antioxidant and antimicrobial compounds from Anacardium occidentale L. (Anacardiaceae) leaf extract. Journal of King Saud University - Science, 27(3), 244–252. https://doi.org/10.1016/j.jksus.2014.12.004

Aoki, Y., Van Trung, N., & Suzuki, S. (2019). Impact of piper betle leaf extract on grape downy mildew: Effects of combining 4-allylpyrocatechol with eugenol, α-pinene or β-pinene. Plant Protection Science, 55(1), 23–30. https://doi.org/10.17221/53/2018-PPS

Ataguba, G. A., Dong, H. T., Rattanarojpong, T., Senapin, S., & Salin, K. R. (2018). Piper betle Leaf Extract Inhibits Multiple Aquatic Bacterial Pathogens and In Vivo Streptococcus agalactiae Infection in Nile Tilapia. Turkish Journal of Fisheries and Aquatic Sciences, 61(3), 1–37. https://doi.org/10.4194/1303-2712-v18

Avijit, B., & Zerin, T. (2020). Comparative Phytochemical and Antibacterial properties of Piper betle Leave Extracts from Barguna and Moheshkhali , Bangladesh. 14(2), 125–132

Baba, S. A., & Malik, S. A. (2015). Determination of total phenolic and flavonoid content, antimicrobial and antioxidant activity of a root extract of Arisaema jacquemontii Blume . Journal of Taibah University for Science, 9(4), 449–454. https://doi.org/10.1016/j.jtusci.2014.11.001

Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71–79. https://doi.org/10.1016/j.jpha.2015.11.005

Bangash, F. A., Hashmi, A., Mahboob, A., Zahid, M., Hamid, B., Muhammad, S., Shah, Z., & Afzaal, H. (2012). In-Vitro Antibacterial activity of Piper Betel Leaf extracts. Journal of Applied Pharmacy, 4(04), 196–203. https://doi.org/10.21065/19204159.4.196

Barbieri, R., Coppo, E., Marchese, A., Daglia, M., Sobarzo-Sánchez, E., Nabavi, S. F., & Nabavi, S. M. (2017). Phytochemicals for human disease: An update on plant-derived compounds antibacterial activity. Microbiological Research, 196, 44–68. https://doi.org/10.1016/j.micres.2016.12.003

Bilal, M., Rasheed, T., Iqbal, H. M. N., Hu, H., Wang, W., & Zhang, X. (2017). Macromolecular agents with antimicrobial potentialities: A drive to combat antimicrobial resistance. International Journal of Biological Macromolecules, 103, 554–574. https://doi.org/10.1016/j.ijbiomac.2017.05.071

Dusane, D. H., Hosseinidoust, Z., Asadishad, B., & Tufenkji, N. (2014). Alkaloids modulate motility, biofilm formation and antibiotic susceptibility of uropathogenic Escherichia coli. PLoS ONE, 9(11), 1–9. https://doi.org/10.1371/journal.pone.0112093

Duval, J., Pecher, V., Poujol, M., & Lesellier, E. (2016). Research advances for the extraction, analysis and uses of anthraquinones: A review. Industrial Crops and Products, 94, 812–833. https://doi.org/10.1016/j.indcrop.2016.09.056

Felhi, S., Daoud, A., Hajlaoui, H., Mnafgui, K., Gharsallah, N., & Kadri, A. (2017). Solvent extraction effects on phytochemical constituents profiles, antioxidant and antimicrobial activities and functional group analysis of Ecballium elaterium seeds and peels fruits. Food Science and Technology, 37(3), 483–492. https://doi.org/10.1590/1678-457x.23516

Guil-Guerrero, J. L., Ramos, L., Moreno, C., Zúñiga-Paredes, J. C., Carlosama-Yepez, M., & Ruales, P. (2016). Antimicrobial activity of plant-food by-products: A review focusing on the tropics. Livestock Science, 189, 32–49. https://doi.org/10.1016/j.livsci.2016.04.021

Harborne. (1984). Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. second ed., Chapman and Hall, New York, USA. In Chapmer and Hall

Hartini, Y. S., Seta Diaseptana, Y. M., Nugraheni Putri, R., & Susanti, L. E. (2018). Antagonistic Antibacterial Effect of Betel and Red Betel Combination against Gram-positive and Gram-negative Bacteria. International Journal of Current Microbiology and Applied Sciences, 7(05), 267–272. https://doi.org/10.20546/ijcmas.2018.705.035

Hidayati, D. N., Hidayati, N., Evinda, E., Fitriana, N. R., & Kusumadewi, A. P. (2019). Antibacterial Activity of Leucaena leucocephala Leaf Extract Ointment against Staphylococcus aureus and Staphylococcus epidermidis. Pharmaciana, 9(1), 175–182. https://doi.org/10.12928/pharmaciana.v9i1.12328

Houghton, P. J., & Raman, A. (1998). Laboratory handbook for the extractation of natural extracts. In Laboratory Handbook for the Extractation of Natural Extracts (1th ed.). International Thompson Publishing. https://doi.org/10.1007/978-1-4615-5809-5

Ibrahim, A. M., Lawal, B., Abubakar, A. N., Tsado, N. A., Kontagora, G. N., Gboke, J. A., & Berinyuy, E. B. (2018). Antimicrobial and free radical scavenging potentials of n-hexane and ethyl acetate extracts of Phyllanthus Fraternus. Nigerian Journal of Basic and Applied Sciences, 25(2), 6. https://doi.org/10.4314/njbas.v25i2.2

Jamil, M., ul Haq, I., Mirza, B., & Qayyum, M. (2012). Isolation of antibacterial compounds from Quercus dilatata L. through bioassay guided extractation. Annals of Clinical Microbiology and Antimicrobials, 11(11), 1–11. https://doi.org/10.1186/1476-0711-11-11

Janmanchi, H., Raju, A., Degani, M. S., Ray, M. K., & Rajan, M. G. R. (2017). Antituberculosis, antibacterial and antioxidant activities of Aegiceras corniculatum, a mangrove plant and effect of various extraction processes on its phytoconstituents and bioactivity. South African Journal of Botany, 113, 421–427. https://doi.org/10.1016/j.sajb.2017.09.019

Junairiah, Ni’matuzahroh, Istighfari, N., Zuraidassanaaz, & Sulistyorini, L. (2018). Isolation and indetification of secondary metabolites of black betel (Piper betle L var nigra). Jurnal Kimia Riset, 3(2), 131–138. https://doi.org/10.1017/CBO9781107415324.004

Kaur, D. C., & Chate, S. S. (2015). Study of antibiotic resistance pattern in methicillin resistant staphylococcus aureus with special reference to newer antibiotic. Journal of Global Infectious Diseases, 7(2), 78–84. https://doi.org/10.4103/0974-777X.157245

Kavitha, S., & Jeevaratnam, K. (2016). Molecular characterization of lactobacilli isolated from Piper betle L. var. Pachaikodi and Comparative Analysis of the Antimicrobial Effects of Isolate Lactobacillus plantarum KJB23 and Betel Leaves Extract. Food Biotechnology, 30(2), 123–136. https://doi.org/10.1080/08905436.2016.1166440

Kusuma, S., Mita, S., & Mutiara, A. (2017). Antimicrobial lotion containing red Piper betle leaf (Piper crocatum Ruiz and Pav) ethanolic extract for topical application. National Journal of Physiology, Pharmacy and Pharmacology, 8(1), 1. https://doi.org/10.5455/njppp.2018.8.1042115112017

María, R., Shirley, M., Xavier, C., Jaime, S., David, V., Rosa, S., & Jodie, D. (2018). Preliminary phytochemical screening, total phenolic content and antibacterial activity of thirteen native species from Guayas province Ecuador. Journal of King Saud University - Science, 30(4), 500–505. https://doi.org/10.1016/j.jksus.2017.03.009

Marín, L., Miguélez, E. M., Villar, C. J., & Lombó, F. (2015). Bioavailability of dietary polyphenols and gut microbiota metabolism: Antimicrobial properties. BioMed Research International, 2015. https://doi.org/10.1155/2015/905215

Meinisasti, R., Muslim, Z., & Sunita, R. (2020). The effectiveness test of Piper Betle Leaf ethanol extract cream ( Piper Betle Linn ) toward propionibacterium acnes bacterial growth. Bioscientia Medicina, 4(2), 10–17

Mgbeahuruike, E. E., Yrjönen, T., Vuorela, H., & Holm, Y. (2017). Bioactive compounds from medicinal plants: Focus on Piper species. South African Journal of Botany, 112, 54–69. https://doi.org/10.1016/j.sajb.2017.05.007

Mostafa, A. A., Al-askar, A. A., Almaary, K. S., Dawoud, T. M., Sholkamy, E. N., & Bakri, M. M. (2018). Saudi Journal of Biological Sciences Antimicrobial activity of some plant extracts against bacterial strains causing food poisoning diseases. Saudi Journal of Biological Sciences, 25(2), 361–366. https://doi.org/10.1016/j.sjbs.2017.02.004

Mufrod, S., & Wahyuono, and S. (2016). Piper betle Leaves Extract Patch: Evaluation Of Antibacterial Activity, Release Profile Of Eugenol, And Local Tolerance. Traditional Medicine Journal, 21(2), 104–110. Https://Doi.Org/10.22146/Tradmedj.12825

Neill, J. I. M. O. (2016). Tackling drug-resistant infections globally : final report and recommendations the review on antimicrobial resistance (Issue May)

Oussalah, M., Caillet, S., & Lacroix, M. (2006). Mechanism of action of Spanish oregano, Chinese cinnamon, and savory essential oils against cell membranes and walls of Escherichia coli O157:H7 and Listeria monocytogenes. Journal of Food Protection, 69(5), 1046–1055. https://doi.org/10.4315/0362-028X-69.5.1046

Pandey, S., Satpathy, G., & Gupta, R. K. (2014). Evaluation of nutritional , phytochemical , antioxidant and antibacterial activity of exotic fruit “ Limonia acidissima .â€3(2), 81–88

Pyla, R., Kim, T., Silva, J. L., & Jung, Y. (2010). International Journal of Food Microbiology Enhanced antimicrobial activity of starch-based fi lm impregnated with thermally processed tannic acid , a strong antioxidant. International Journal of Food Microbiology, 137(2–3), 154–160. https://doi.org/10.1016/j.ijfoodmicro.2009.12.011

Singh, T. P., Chauhan, G., Agrawal, R. K., & Mendiratta, S. K. (2019). In vitro study on antimicrobial, antioxidant, FT-IR and GC–MS/MS analysis of Piper betle L. leaves extracts. Journal of Food Measurement and Characterization, 13(1), 466–475. https://doi.org/10.1007/s11694-018-9960-8

Stahl, E. (1969). Thin-Layer Chromatography A Laboratory Handbook. In Journal of Chemical Information and Modeling (Issue 2). Springer Berlin Heidelberg. https://doi.org/10.1007/978-3-642-88488-7

Su, P. W., Yang, C. H., Yang, J. F., Su, P. Y., & Chuang, L. Y. (2015). Antibacterial activities and antibacterial mechanism of polygonum cuspidatum extracts against nosocomial drug-resistant pathogens. Molecules, 20(6), 11119–11130. https://doi.org/10.3390/molecules200611119

Swamy, M. K., Akhtar, M. S., & Sinniah, U. R. (2016). Antimicrobial properties of plant essential oils against human pathogens and their mode of action: An updated review. Evidence-Based Complementary and Alternative Medicine, 2016, 1–21. https://doi.org/10.1155/2016/3012462

Syahidah, A., Saad, C. R., Hassan, M. D., Rukayadi, Y., Norazian, M. H., & Kamarudin, M. S. (2017). Phytochemical analysis, identification and quantification of antibacterial active compounds in betel leaves, piper betle methanolic extract. Pakistan Journal of Biological Sciences, 20(2), 70–81. https://doi.org/10.3923/pjbs.2017.70.81

Tong, S. Y. C., Davis, J. S., Eichenberger, E., Holland, T. L., & Fowler, V. G. (2015). Staphylococcus aureus infections: Epidemiology, pathophysiology, clinical manifestations, and management. Clinical Microbiology Reviews, 28(3), 603–661. https://doi.org/10.1128/CMR.00134-14

Upadhyay, A., Upadhyaya, I., Kollanoor-Johny, A., & Venkitanarayanan, K. (2014). Combating Pathogenic Microorganisms Using Plant-Derived Antimicrobials: A Minireview of the Mechanistic Basis. BioMed Research International, 2014. https://doi.org/10.1155/2014/761741

Valle, D. L., Cabrera, E. C., Puzon, J. J. M., & Rivera, W. L. (2016). Antimicrobial activities of methanol, ethanol and supercritical CO2 extracts of philippine Piper betle L. on clinical isolates of Gram positive and Gram negative bacteria with transferable multiple drug resistance. PLoS ONE, 11(1), 1–14. https://doi.org/10.1371/journal.pone.0146349

Waghmode, S., Pawar, S., Kalyankar, V., Dhamangaonkar, B., Dagade, S., & Cukkemane, A. (2017). Biochemical profiling of antifungal activity of betel leaf(Piper betle L.) extract and its significance in traditional medicine. Journal of Advanced Research in Biotechnology, 2(1), 1–4. https://doi.org/10.15226/2475-4714/2/1/00116

Wang, C., Chen, H., Wu, Z., Jhan, Y., Shyu, C., & Chou, C. (2016). Antibacterial and Synergistic Activity of Pentacyclic Triterpenoids Isolated from Alstonia scholaris. Molecules MDPI, 21, 1–11. https://doi.org/10.3390/molecules21020139

Downloads

Published

2021-07-30

Issue

Vol. 11 No. 2 (2021): Pharmaciana

Section

Biology Pharmacy

License

Authors who publish with Pharmaciana agree to the following terms:

  1. Authors retain copyright and grant the journal the right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (CC BY-SA 4.0) that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal. 
  2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
  3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.