Metabolite Detection and Antibacterial Activity of Fungal Endophytic Extracts Isolated from Brotowali (Tinospora crispa) Plants using TLC-Bioautography Assay

Mangosteen leaves have flavonoid compounds, tannins, and saponins that can be efficacious as antibacterial. The aim of this study was to determine the fraction of mangosteen leaves having an antibacterial effect and knowing the class of compounds identified as having antibacterial activity after TLC- Bioautography testing was carried out. The samples were extracted using 96% maceratarion method and fractioned using liquid-liquid fractionation method with methanol, n-hexane and ethyl acetate solvents, antibacterial activity using agar diffusion method (Kirby and Bauer) with 3 concetrations namely 10%, 20% and 30%. Thin Layer Chromatography (TLC) uses n-hexane and chloroform solvens. TLC-Bioautography uses contact bioautography methods. The resultd showed that mangosteen leaves in methanol fraction with a concentration of 30% had a very large inhibitory activity again Staphylococcus aureus and ethyl acetate fraction with a concentration of 30% had the gratest antibacterial activity against Escherichia coli. The results of the TLC- Bioautography study showed that the flavonoids compounds after spraying with AlCl3 and the mangosteen leaf Biosutography test had inhibitory zone activity against the bacteria Staphylococcus aureus dan Escherichia coli. Keywords: Mangosteen Leaves. Antibacterial, TLC Bioautography. ABSTRAK Daun manggis mempunyai senyawa flavonoid, tanin, dan saponin yang dapat berkhasiat sebagai antibakteri. Penelitian ini bertujuan untuk mengetahui fraksi daun manggis memiliki efek antibakteri dan mengetahui golongan senyawa yang teridentifikasi memiliki aktivitas antibakteri setelah dilakukan pengujian KLT Bioautografi. Sampel diektrak dengan metode maserasi dengan pelarut 96% dan difraksinasi dengan metode fraksinasi cair-cair dengan pelarut metanol, n-heksan dan etil asetat, aktivitas antibakteri menggunakan metode difusi agar (Kirby and Bauer) dengan 3 kosentrasi yaitu 10%, 20% dan 30%. Kromatografi Lapis Tipis (KLT) menggunakan pelarut n-heksan dan klorofom. KLT-Bioautografi menggunakan metode bioautografi kontak. Hasil penelitian menunjukan daun manggis pada fraksi metanol dengan kosentrasi 30% memiliki aktivitas zona hambat ppaling besar terhadapat bakteri Staphylococcus aureus dan fraksi etil asetat dengan kosentrasi 30% memiliki aktivitas antibakteri paling besar terhadap Escherichia coli. Hasil penelitian KLT-Bioautografi menunjukan golongan senyawa flavonoid setelah disemprotkan dengan AlCl3 dan uji Bioautografi daun manggis memiliki aktivitas zona hambat terhadap bakteri Staphylococcus aureus dan Escherichia coli. Kata kunci : Daun Manggis, Antibakteri, KLT- Bioautografi.

Download Full-text

ANTIBACTERIAL ACTIVITY OF ROBUSTA COFFEE (Coffea robusta L.) PEEL EXTRACT AGAINST HUMAN PATHOGENIC BACTERIA

Journal of Experimental Biology and Agricultural Sciences ◽

10.18006/2021.9(spl-2-icopmes_2020).s264.s268 ◽

2021 ◽

Vol 9 (Spl-2-ICOPMES_2020) ◽

pp. S264-S268

Author(s):

Herlina Rante ◽

◽

Subehan . ◽

Retno Wulandari ◽

Yayu Mulsiani Evary ◽

...

Keyword(s):

Antibacterial Activity ◽

Infectious Diseases ◽

Pathogenic Bacteria ◽

Fruit Peel ◽

E Coli ◽

Robusta Coffee ◽

Human Pathogenic Bacteria ◽

Tlc Bioautography ◽

Coffea Robusta ◽

Peel Extract

Now in these days infectious diseases seriously affect human health and sometimes these infections might become the cause of human mortality. Most of these infectious diseases are caused by bacteria, viruses, and fungi. Although large numbers of antibiotics are available increasing drug resistance in these microorganisms became a serious matter of concern in the scientific community. There is an urgent need for research on alternate natural products that can manage these pathogenic microorganisms without inducing any resistance. The purpose of this study was to determine the antibacterial activity of Robusta coffee (Coffea robusta L.) fruit peel extract against 5 human pathogenic bacteria i.e. Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Salmonella thypi NCTC 786. The sample was extracted using the maceration method with methanol as the solvent. The antibacterial activity of fruit peel extract was determined by using the agar diffusion method while the presence of active ingredients was determined by the using TLC-Bioautography assay performed using the mobile phase of n-hexane: ethyl acetate (1 : 3). The results of the study revealed significant antibacterial activity of coffee peel extract against E. coli and B. subtilis with an inhibition zone of 10.15 mm and 10.96 mm, respectively. Furthermore, results of the TLC-Bioautography revealed that the compounds at Rf 0.76 inhibit the growth of E. coli and the compounds at Rf 0.27 inhibit the growth of B. subtilis bacteria. These active spots were suspected to be flavonoid and phenolic compounds, respectively but further confirmation detail study is required in the future.

Download Full-text

ANTIBACTERIAL ACTIVITY AND TLC-BIOAUTOGRAPHY ANALYSIS OF THE ACTIVE FRACTIONS OF Muntingia calabura L. LEAVES AGAINST Staphylococcus aureus

Journal of Pharmaceutical Sciences and Community ◽

10.24071/jpsc.002362 ◽

2020 ◽

Vol 17 (2) ◽

pp. 69-75

Author(s):

Rizky H. Mawardi ◽

Nanik Sulistyani ◽

Nurkhasanah Nurkhasanah ◽

Ricke Desyratnaputri

Keyword(s):

Staphylococcus Aureus ◽

Antibacterial Activity ◽

Ethyl Acetate ◽

Inhibition Zone ◽

Ethyl Acetate Fraction ◽

Active Fraction ◽

Methanol Fraction ◽

Zone Diameter ◽

Tlc Bioautography ◽

Muntingia Calabura

This study aims to determine the antibacterial activity and TLC-bioautography profile of the active fractions of Muntingia calabura L. leaves against Staphylococcus aureus. Muntingia calabura L. leaves were macerated with ethanol 96% then fractionated with n-hexane, ethyl acetate, and methanol solvent, respectively. The antibacterial activity was tested by the Kirby-Bauer method to determine the most active fraction and the lowest concentration that inhibited the growth of Staphylococcus aureus. TLC-bioautography was tested using chloroform: ethyl acetate (2:8) as the mobile phase and silica gel F254 as the stationary phase. Antibacterial activity test of n-hexane and ethyl acetate fractions at a concentration of 10% w/v showed activities with inhibition zone diameter of 0.33±0.288 and 9.66±5.77 mm, respectively. At the same time, the methanol fraction showed no activity. The lowest concentration of ethyl acetate fraction which still showed the inhibition zone was 0.312% w/v. The TLC-bioautography profile showed active spots with an Rf value of 0.82 and had an inhibitory zone diameter of 4.013±0.864 mm. It can be concluded that the ethyl acetate fraction was the most active fraction that inhibited the growth of Staphylococcus aureus and had one active spot on the bioautography test.

Download Full-text

UJI AKTIVITAS ANTIBAKTERI EKSTRAK METANOL DAUN KIRINYUH (Chromolaena odorata) TERHADAP BAKTERI MDR (Multi Drug Resistant) DENGAN METODE KLT BIOAUTOGRAFI

JURNAL BIOSAINS ◽

10.24114/jbio.v6i2.16600 ◽

2020 ◽

Vol 6 (2) ◽

pp. 45

Author(s):

Nur Hasanah ◽

Endang Sulistyarini Gultom

Keyword(s):

Antibacterial Activity ◽

Methanol Extract ◽

Methanolic Extract ◽

Ammonia Vapor ◽

Test Results ◽

Chromolaena Odorata ◽

Tlc Bioautography ◽

Rf Values ◽

Inhibition Zones ◽

Spray Reagents

ABSTRAK Penelitian ini bertujuan untuk mengetahui aktivitas antibakteri ekstrak metanol daun kirinyuh (Chromolaena odorata) terhadap bakteri MDR dengan metode KLT-bioautografi. Bakteri uji yang digunakan adalah bakteri MDR S.lugdunensis MRSA, K.pneumoniae ESBL dan P.aeruginosa ESBL. Metode ekstraksi dilakukan dengan cara maserasi menggunakan pelarut metanol p.a.Untuk pengujian KLT-bioautografi, ekstrak metanol daun kirinyuh ditotolkan pada plat KLT kemudian dielusi dengan eluen metanol:kloroform (9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8 dan 1:9), kemudian plat KLT ditempelkan pada media MHA yang telah diinokulasikan ke 3 bakteri MDR dan diinkubasi selama 24 jam. Zona hambat yang terbentuk dilakukan identifikasi senyawa menggunakan pereaksi semprot FeCl3, dragendorff, uap amoniak, SbCl3 dan Lieberman bouchardat. Hasil uji KLT-bioautografi menunjukkan bahwa ekstrak metanol daun kirinyuh dapat menghambat pertumbuhan bakteri yang ditandai terbentuknya zona bening pada EMDKF3, EMDKF7b, EMDKF9b dengan masing-masing nilai Rf 0,47 cm (fenol), 0,87 cm (flavonoid), 0,92 cm (flavonoid) pada bakteri S.lugdunensis MRSA dan EMDKF2, EMDKF7a, EMDKF7c dengan masing-masing nilai Rf 0,6 cm (flavonoid), 0,57 cm (alkaloid), 0,96 cm (flavonoid) pada bakteri K.pneumoniae ESBL, sedangkan pada bakteri P.aeruginosa ESBL ekstrak metanol daun kirinyuh tidak memiliki aktivitas antibakteri yang ditunjukkan dengan tidak ada zona bening yang terbentuk. Senyawa yang berperan sebagai antibakteri terhadap bakteri S.lugdunensis MRSA dan K.pneumoniae ESBL adalah senyawa golongan alkaloid, flavonoid dan fenol. Kata Kunci: Chromolaena odorata, Bakteri MDR, Aktivitas Antibakteri, KLT Bioautografi ABSTRACT This study aims to determine the antibacterial activity of methanol extract of kirinyuh leaves (Chromolaena odorata) against MDR bacteria by TLC-bioautography method. The test bacteria used were MDR S.lugdunensis MRSA, K.pneumoniae ESBL and P.aeruginosa ESBL. The extraction method is carried out by maceration using methanol solvent p.a. for TLC-bioautographic testing, the methanol extract of the kirinyuh leaves was poured on the TLC plate then eluted with methanol:chloroform eluent (9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8 and 1:9), then the TLC plate was attached to the MHA media which had been inoculated to 3 MDR bacteria and incubated for 24 hours. Inhibition zones formed were identified using compound spray reagents FeCl3, dragendorff, ammonia vapor, SbCl3 and Lieberman bouchardat. The TLC-bioautographic test results showed that the methanolic extract of kirinyuh leaves can inhibit bacterial growth marked by the formation of clear zones in EMDKF3, EMDKF7b, EMDKF9b with each Rf value of 0,47 cm (phenol), 0,87 cm (flavonoids), 0,92 cm (flavonoids) in S.lugdunensis MRSA and EMDKF2, EMDKF7a, EMDKF7c bacteria with Rf values of 0,6 cm (flavonoids), 0,57 cm (alkaloids), 0,96 cm (flavonoids) in K.pneumoniae ESBL, whereas in P.aeruginosa ESBL bacteria the methanolic extract of kirinyuh leaves had no antibacterial activity as indicated by no clear zone formed. The compounds that act as antibacterial against the bacteria S.lugdunensis MRSA and K.pneumoniae ESBL are alkaloid, flavonoid and phenol compounds.Keywords: Chromolaena odorata, MDR Bacteria, Antibacterial Activity, TLC Bioautography

Download Full-text

Diversity of endophytic fungi from Vernonia amygdalina, their phenolic and flavonoid contents and bioactivities

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d210202 ◽

2020 ◽

Vol 21 (2) ◽

Author(s):

Praptiwi Praptiwi ◽

Ahmad Fathoni ◽

Muhammad Ilyas

Keyword(s):

Antioxidant Activity ◽

Antibacterial Activity ◽

Endophytic Fungi ◽

Antioxidant Activities ◽

Activity Index ◽

Total Phenolic ◽

Vernonia Amygdalina ◽

Phenolic Contents ◽

Tlc Bioautography ◽

Phenolic And Flavonoid

Abstract. Praptiwi, Fathoni A, Ilyas M. 2019. Diversity of endophytic fungi from Vernonia amygdalina, their phenolic and flavonoid contents and bioactivities. Biodiversitas 20: 436-441. There are nine endophytic fungi in total associated with stem and leaves of Vernonia amygdalina were determined for their total phenolic and flavonoids content as well as assessed for their antibacterial and antioxidant activities. Total phenolic and flavonoid contents were determined by spectrophotometric methods. The antibacterial activity was performed using Thin Layer Chromatography (TLC) - Bioautography assay against S.aureus and E.coli. The antioxidant activity was carried out using TLC- Bioautography by DPPH scavenging assay. Broth serial microdilution assay was used to determine the minimum inhibitory concentrations (MIC) and the half-maximum inhibitory concentration (IC50). Among the identified isolates, there was one isolate of Phomopsis sp., two isolates of Phoma sp., two isolates of Lasiodiplodia sp., two isolates of Colletotrichum sp. and two isolates of Phyllosticta sp. The results showed that the phenolic contents were ranging from 0.53-116.03 mg GAE/g extract, flavonoids contents were ranging from 81.12 to 390.21 mg quercetin equivalent (QE)/g extract. MICs of endophytic fungi against S.aureus and E.coli ranged from 256 to >256 µg/mL and >256 µg/mL respectively that categorized as moderate antibacterial activity. The antioxidant activity of endophytic fungi associated with V. amygdalina was >128 µg/mL, and the antioxidant activity index (AAI) was 0.24.

Download Full-text

Uji Aktivitas Antibakteri Propionibacterium acnes Ekstrak Etanol dan Fraksi Daun Karuk (Piper sarmetosum Roxb. Ex. Hunter) serta Analisis KLT Bioautografi

Jurnal Riset Farmasi ◽

10.29313/jrf.v1i1.26 ◽

1970 ◽

Vol 1 (1) ◽

pp. 9-15

Author(s):

Anisa Dwi Nuraeni ◽

Yani Lukmayani ◽

Reza Abdul Kodir

Keyword(s):

Antibacterial Activity ◽

Ethyl Acetate ◽

Propionibacterium Acnes ◽

Ethanol Extract ◽

Ethyl Acetate Fraction ◽

Hexane Fraction ◽

Water Fraction ◽

Liquid Liquid Extraction ◽

Tlc Bioautography ◽

Ethanol Extracts

Abstract. Karuk (Piper sarmetosum Roxb. Ex. Hunter) is a plant that is considered to have antibacterial activity. Acne is a problem in the skin caused by the bacteria Propionibacterium acnes. This research’s aimed to determine the potential antibacterial activity of ethanol extracts and karuk leaf’s fractions, and can find out the class of chemical compounds contained from the ethanol extract and the fraction of karuk leaves which have antibacterial activity. Karuk leaf was extracted by maceration method using 96% ethanol solvent, followed by fractionation using the liquid-liquid extraction method with n-hexane, ethyl acetate, and water solvents. Antibacterial activity test of ethanol extract and fraction with a concentration of 2%, 4%, 6%, and 8% using the well diffusion agar method. To identify the class of compounds that have antibacterial activity, the TLC Bioautography method is used. From the results of research that has been done shows that ethanol extract and n-hexane fraction can inhibit the growth of Propionibacterium acnes with the highest inhibitory diameter of 10.11mm in the extract, and in the n-hexane fraction of 10.93 mm. in the ethyl acetate fraction and the water fraction there is no inhibitory zone formed so that it cannot provide antibacterial activity. The TLC Bioautography results did not show any inhibitory zones and there was no change in color when given a spotting viewer. So that compounds cannot be identified that have antibacterial activity Abstrak. Karuk (Piper sarmetosum Roxb. Ex. Hunter) merupakan tanaman yang diketahui memiliki aktivitas sebagai antibakteri. Jerawat merupakan permasalahan pada kulit disebabkan adanya bakteri Propionibacterium acnes. Penelitian ini bertujuan untuk menentukan potensi aktivitas antibakteri dari ekstrak etanol dan fraksi daun karuk, serta dapat mengetahui golongan senyawa kimia yang terkandung dari hasil ekstrak etanol dan fraksi daun karuk yang memiliki aktivitas antibakteri. Daun karuk diekstraksi dengan metode maserasi menggunakan pelarut etanol 96%, dilanjutkan dengan fraksinasi menggunakan metode ekstraksi cair-cair dengan pelarut n-heksan, etil asetat, dan air. Uji aktivitas antibakteri ekstrak etanol dan fraksi dengan konsentrasi 2%, 4%, 6%, dan 8% menggunakan metode difusi sumuran agar. Untuk mengetahui golongan senyawa yang memiliki aktivitas sebagai antibakteri digunakan metode KLT Bioautografi. Dari hasil penelitian yang telah dilakukan menunjukan bahwa ekstrak etanol dan fraksi n-heksan dapat menghambat pertumbuhan Propionibacterium acnes dengan diameter hambat tertinggi 10,11mm pada ekstrak, dan pada fraksi n-heksan 10,93 mm. pada fraksi etil asetat dan fraksi air tidak terbentuk adanya zona hambat sehingga tidak dapat memberikan aktivitas sebagai antibakteri. Hasil KLT Bioautografi tidak menunjukan adanya zona hambat serta tidak terdapat perubahan warna ketika diberikan penampak bercak. Sehingga tidak dapat teridentifikasi senyawa yang memiliki aktivitas antibakteri.

Download Full-text

Detection of antibacterial activity of essential oil components by TLC-bioautography using luminescent bacteria

JPC - Journal of Planar Chromatography - Modern TLC ◽

10.1556/jpc.26.2013.2.2 ◽

2013 ◽

Vol 26 (2) ◽

pp. 114-118 ◽

Cited By ~ 9

Author(s):

Györgyi Horváth ◽

Béla Kocsis ◽

Éva Lemberkovics ◽

Andrea Böszörményi ◽

Péter Ott ◽

...

Keyword(s):

Antibacterial Activity ◽

Essential Oil ◽

Tlc Bioautography ◽

Luminescent Bacteria ◽

Oil Components

Download Full-text

AKTIVITAS ANTIBAKTERI DAN ANALISIS KLT-BIOAUTOGRAFI DARI FRAKSI BIJI KOPI ROBUSTA (Coffea canephora Pierre ex A. Froehner)

PHARMACON ◽

10.35799/pha.8.2019.29325 ◽

2019 ◽

Vol 8 (3) ◽

pp. 516

Author(s):

Wazhifa Andarini Paputungan ◽

Widya Astuty Lolo ◽

Jainer Pasca Siampa

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Antibacterial Activity ◽

Ethanol Extract ◽

Inhibition Zone ◽

Coffea Canephora ◽

Methanol Fraction ◽

Robusta Coffee ◽

Tlc Bioautography ◽

Coffee Beans

Coffee is a plant that contains compounds such as flavonoids, which have antibacterial activity. The purpose of this study was to determine the fraction of the ethanol extract of robusta coffee beans whether it has an antibacterial effect and know the class of compounds identified after TLC-Bioautography. The extraction method used was maceration with 96% ethanol and fractionation using liquid-liquid method with methanol, n-hexane, and ethyl acetate. Antibacterial testing using the agar diffudion method with concentration of 10%, 20%, 30% respectively. The results of study of 30% methanol fraction for Staphylococcus aureus bacteria have a 12,58 mm inhibition zone and Escherichia coli bacteria 11,60 in the strong category. The largest fraction of the inhibition zone is the 30% methanol fraction of robusta coffee beans was carried out by TLC-Bioautography test using the mobile phase of chloroform : n-hexane (1:2). Tests for antibacterial activity with contact bioautography methods resulted in spotting on the chromatogram, which produces a inhibitory zone. The characteristics of the spots were carried out with the appearance of AlCl3 and dragendorf patches which were suspected to be flavonoids and alkaloids. Keywords : Fraction, Antibacterial, Coffea canephora Pierre ex A. Froehner, TLC Bioautography. ABSTRAK Kopi adalah tanaman yang mengandung senyawa antara lain flavonoid yang mempunyai aktivitas antibakteri. Tujuan dari penelitian ini adalah untuk mengetahui fraksi dari ekstrak etanol biji kopi robusta apakah memiliki efek antibakteri dan mengetahui golongan senyawa yang teridentifikasi memiliki aktivitas antibakteri setelah dilakukan pengujian KLT-Bioautografi. Metode ekstraksi yang digunakan adalah maserasi dengan pelarut etanol 96% dan dilakukan fraksinasi dengan metode cair-cair dengan pelarut metanol, n-heksan, dan etil asetat. Pada pengujian antibakteri menggunakan metode difusi agar dengan konsentrasi 10%, 20%, 30%. Hasil penelitian fraksi metanol konsentrasi 30% untuk bakteri Staphylococcus aureus memiliki zona hambat 12,58 mm dan bakteri Escherichia coli 11,60 mm dengan kategori kuat. Fraksi yang paling besar zona hambatnya yaitu fraksi metanol konsentrasi 30% dilakukan pengujian KLT-Bioautografi menggunakan Bioautografi kontak. Aktivitas antibakteri terhadap fraksi metanol biji kopi robusta dilakukan dengan uji KLT-Bioautografi menggunakan fase gerak kloroform : n-heksan (1:2). Pengujian aktivitas antibakteri dengan metode bioautografi kontak hasil yang diperoleh terdapat bercak pada kromatogram yang menghasilkan zona hambat. Karakteristik bercak dilakukan dengan penampak bercak AlCl3 dan dragendorf diduga bercak tersebut adalah flavonoid dan alkaloid. Kata kunci : Fraksi, Antibakteri, Coffea canephora Pierre ex A. Froehner, KLT-Bioautografi.

Download Full-text