ANTIMICROBIAL ACTIVITIES OF TERMINALIA MUELLERI BENTH. LEAVES, COMBRETACEAE, AND IT’S ACTIVE COMPOUNDS

• Main Author: Anam, Khairul
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45853
• Institution: Institut Teknologi Bandung

One of foci attention in health topic is the continuous supply of anti-infection medicine to overcome infection diseases, which is the third biggest cause of mortality in the world. Variety of this medicine is limited while some patogenic micro-organisms are resistant to them. Related to this issue, research in finding new variety of medicines is needed, especially some plants species traditionally used in treating some diseases, including infection disease. Indonesian society has traditionally used plants in treating several kinds of diseases including infection, but there are not many evidence base of the usage. Meanwhile, the information of the plants usage in traditional medication is considered as one of the approaches in finding new medicine variety and the National Research Council supports the development of natural medicine by using Indonesia’s biological resources. Therefore, this research, that looks for new antimicrobial medicine from Indonesia’s plants, is very relevant. Terminalia catappa L. is one of varieties that is grown widely in Indonesia and traditionally used as medicine. Like the others member of Combretaceae family, this plant is also reported to have antimicrobial activities. In previous research, it was known that T. muelleri Benth. have the strongest antimicrobial activities than the 17 other species of Combretacea family, grown in Indonesia. There are only a few researches in T. muelleri species, therefore it would be a valuable evaluation for getting potential antimicrobial medicine. The aim of this research is to verify using T. muelleri as antimicrobial The specific aims of this research are to isolate the antimicrobial compounds, identify the chemical structure of the active compounds, and to study the potential of antimicrobial activity and its mode of action. v Strategies used in this research is through chemistry dan antimicrobial activity approaches. Isolation of the active compound from T. muelleri leaves was conducted directly with antimicrobial test guidance. Antimicrobial test was held by the agar diffussion method with Escherichia coli ATCC 25923, Staphylococcus aureus ATCC 25922 and fungi Candida albicans ATCC 10132. The potential of antimicrobial activity was studied by the determination of the minimum inhibitory concentration (MIC) against various bacteria and fungi, and their equaivalency with the referred antibiotic. Mode of antimicrobial action of the isolate was studied by observing its effect on microbe morphology using Scanning Electron Microscope (SEM) and Transmission Electron microscope (TEM). Parameter being observed in SEM product is the change of shape or cell morphology compared to the control group, whereas in TEM is the completeness and the damage of the cell component as well the damage pattern of the cell wall. The active isolate was characterized by spectroscopy and the chemistry structure was elucidated. Based on these tests, it was known that ethylacetic extract was more active on the tested microbes than the n-hexane and methanol extract. Fractination of ethylacetic extract was carried out by vacuum liquid chromatography, the stationary phase used was silica gel 60H and the mobile phase were n-hexane, methylenchloride, ethylacetate and methanol, successively in gradient polarity. The fractions were tested for their antimicrobial activities and it was known that EH fraction had the strongest activities on S. aureus bacteria. The isolation of the active compounds of EH fraction was guided by bioautography method and it produced EHJ isolate. The EHJ isolate inhibit the growth of S. aurues (MIC 0.735 mg/ml) and Methicilin-Resistant Staphylococcus aureus (MIC 2.5 mg/ml), but it did not inhibit the growth of positive Gram bacteria B. subtilis, S. lutea, and negative Gram E. coli, and also fungi C. albicans. The activity of 1 mg isolat EHJ was equal to 0.1396 µg tetracycline HCl against S. aureus and 0.6455 µg against Methicilin-Resistant Staphylococcus aureus, also equals to 40.6035 µg peniciline G against S. aureus and equals to 2.9823 and 2.1213 µg vancomycin HCl against S. aureus and Methicilin-Resistant Staphylococcus aureus, respectively. SEM and TEM were used the mode of action of the EHJ isolate on S. aureus cell wall. Antibacterial activities in SEM observation was shown by wrinkle and pore appearance, and imperfectly built cell’s wall. In TEM, it was observed that the damage was on bacteria cells’ wall and nukleoide. Some bacteria have had a thin cell wall, and moreover have not had cell wall. Another bacteria had a thicker than normal cell. Thicking of cell wall is responding system in order to defence bactery’s body from antibiotic that inhibit the cell wall synthesis. The same damages images were shown by SEM and TEM of MRSA. These damage colud be showed on surface structure and bacteria cell component explained that antibacterial activity of EHJ isolate as antibacterial by inhibit the cell wall synthesis. vi The EHJ isolate was characterized by the melting point of 249.6 – 251 o C (without correction), Rf: 0.76 on TLC silica gel 60 HF254 using mobile phase : ethylacetic – formic acid – water (18:1:1) and Rf : 0,65 on TLC silica gel RP-18, using mobile phase: chloroform-methanol (8:2). From the result of characterization using UV-Vis and Infra Red (IR) spectrophotometry, Nuclear Magnetic Resonance (NMR) and Mass spectrometry (MS), it was known that EHJ isolate is 3,4,5-trihydroxybenzoic acid and also known as gallic acid. This research examined phytochemistry and bioactivity aspect of T. muelleri Benth leaves. The characteristics of T. muelleri Benth leaves was’nt graft yet in Materia Media Indonesia monograph, so the result of characteritation from this research could be developed as input for MMI ordering furthemore. Gallic acid of T. muelleri Benth. Leaves for the first time has reported and as one of antimicrobial compounds in these plant. Mode of action of gallic acid on S. aureus which was examined through their effect on damaging of S. aureus cell morphology using SEM and TEM for the first time would be reported on this research The results of this research could be used as evidence base for T. muelleri usage for an alternative therapy against infection diseases. Moreover, it opens up an opportunity for a continuous research in order to identify other antimicrobial compounds which have not been isolated yet from the leaves of T. muelleri, and to develop the isolation technique as well as the genetic engineering to produce active compounds. The antimicrobial mechanism can be studied in advance by the interaction approach with receptor or labelling with radioactive marker compounds.