ISOLATION AND IDENTIFICATION OF ANTIFUNGAL (Candida albicans) COMPOUND FROM THE HULL OF DELIMA FRUITS (Punica granatum L.)

The hull of delima fruits (Punica granatum L.) are traditionally used to cure dysmenorhoe. Preliminary study indicated that extract of the hull was able to inhibit the growth of Candida albicans. Therefore this study was aimed to isolate and identify active compounds responsible for the activity from the hull of P. granatum. The study was initiated by extracting the powdered material with petroleum ether followed by methanol. Antifungal activity test (100 mg/ml) indicated that the petroleum ether extract was more active than the methanol extract (inhibition zone: 10.59 vs 6.92 mm). The pet. ether extract was triturated by n-hexane to give n-hexane insoluble and n-hexane soluble fractions. The latter that was active (inh. zone: 9.50 vs 0.00 mm) was fractionated by vacuum liquid colomn chromatography (vlc; SiO2, n-hexane with increasing amount of ethylacetate) to give 7 fractions (F1-F7). Fraction 2 (inh. zone: 9.05 mm) and 3 (11.05 mm) displayed antifungal activity, then F3 was subjected to contact bioautography to give 2 active compounds [Rf. 0.50 (major) and 0.10 (minor)]. Preparative tlc [SiO2 F-254 nm; nhexane: ethylasetate 4-1, developed 2x) of F3 was aimed to separate 2 active compounds. Due to limited amount of the minor compound, the MFC was applied only for the major compound (Rf. 0.50; 200 mg/ml). The structure identification was done by mean of spectroscopic methods (uv, ir, ms and nmr) to be a setrol type of compound having stigmastane skeleton, esterified by a long-chain fatty acid.Keywords: Punica granatum L., Candida albicans, active compound, terpenoi

Identifikasi dua asam triterpen yang terdapat di dalam amsonia grandiflora, Fam. Apocynaceae

ABSTRAK Amsonia grandiflora adalah salah satu tanaman yang mudah tumbuh didaerah kering. Tanaman ini mengandung karbohidrat dalam jumlah besar disamping mempunyai nilai nutrisi yang sama dengan rumput alfalfa secara in-vitro. Tetapi tanaman ini tidak dimakan oleh ternak yang hidup didaerah tersebut. Dua senyawa asam triterpen dalam tanaman dapat diisolasi dart sari diklormetana A. grandiflora yang tidak larut dalam n-heksana. Berdasarkan alas spektrum inframerah, spektrum massa, spektrum resonansi magnetik inti, titik lebur dan rotasi oplik, identitikasi kedua senyawa tersebut mengarah kepada asam betulinat dan asam oleanolat. Perbandingan secara langsung kedua senyawa tersebut dengan senyawa asam betulinat dan asam oleanolat baku, maka kedua senyawa tersebut disimpulkan sebagai asam betulinat dan asam oleanolat. Keberadaan dua triterpene asam dalam jumlah relatif tinggi memungkinkan mempengaruhi rasa tanaman, yang memungkinkan ternak tidak mau memakannya. Kata kuncl: Amsonia grandiflora, makanan ternak, asam betulinat, asam oleanolat. ABSTRACT An indigenous desert plant, Amsonia grandiflora, has a high carbohydrate content and an animal feed nutritive value in-vitro similar to alfalfahowever, this plant is not grazed in the wild. Two triterpene acids were isolated from the dichloromethane extract of A. grandiflora that was insoluble in n-hexane. Based on their infrared, mass and nuclear magnetic resonance spectra as well as their melting points and optical rotations, the two acids were suspected to be betulinic acid and oleanolic acid. Direct comparison of these two acids with authentic betulinic and oleanolic acid, these acids were confirmed as betulinic acid and oleanolic acid. The high concentrations of oleanolic acid and betulinic acid might impart a sour taste to the plant. This could possibly be the reason that animals in the wild do not grazed this plant. Key words : Amsonia grandiflora, animal feedstocks, betulinic acid, oleanolic acid

Identifikasi lupeol, asetil lupeol dan lupeil t3-hidroksioktadekanoat yang terdapat di dalam fraksi non-polar amsonia grandiflora(fam. Apocynaceae)

ABSTRAK Sebagai kelanjutan penelitian mengenai kandungan kimia dari tanatnan Arnsonta grandiflora, telah diisolasi tiga senyawa dari sari non polar dari sari diklormetana tanaman tersebut. Ketiga senyawa tersebut merupakan senyawa triterpena yang mempunyai kerangka lupan dan diidentifikasi sebagai Lupeol, asetil lupeol dan lupeil /1-hidrolcsioktadekanoat. Identifikasi senyawa didasarkan atas data spektra ms, nmr), don perbandingan langsung dengan lupeol baku. Kata kunci : Amsonia grandiflora, Apocynaceae, lupeol, asetil lupeol dan lupeil Ahidroksioktadekanoate ABSTRACT A phytochemical investigation on the non-polar fraction of Amsonia grandiflora has been done. A triterpene compound of lupane skeleton and its derivatives were isolated. These compounds were identified as lupeol, acetyl lupeol and lupeol /3-hydroxyoctadecanoate based on their chemical and physical data (ir, ms, nmr), and direct comparison with authentic lupeol. Key words: Amsonia grandiflora, Apocynaceae, lupeol, acetyl lupeol and lupeol ft-hydroxyoctadecanoat

Cytotoxicity and antimicrobial test of the bioactive compound isolated from Stylissa flabelliformis sponge

A research of cytotoxicity and antimicrobial test of bioactive compound of Stylissa flabelliformis sponge have been done. Cytotoxicity test were conducted on myeloma cells, while antimicrobial test were conducted against Staphylococus aureus,Eschericia coli and Candida albican. The research was initiated with the isolation of bioactive compound from the active fraction of sponge S. Stylissa monitored by Brine shrimp Lethality tes (BST). The active fraction was chromatograped with silica gel as its immobile phase and the mixture of hexane and ethyl acetate (1:1 ) as the mobile phase to obtain compound 1,2,3 and 4.Compound 3 has the most toxic character because it could kill A. salina equal to 100% at dose 25 mg/ml, and has LC50 of 0,9 mg/ml. Cytotoxicity test with direct staining method using tripan blue on myeloma cell at density of 4,5x10 cell / 100.ul showed that the compound 3 had a high activity against myeloma cell, having LC50 equal to 0,08 mg/ml.The result of antimicrobial effect showed that compound 3 was a fungicide against Candida albicans but having no activity on Staphylococcus aureus and Eschericia coliKey words: sponge, Stylissa flabelliformis , cytotoxicity, fungicid

Skrining toksisitas dengan menggunakan Brine Shrimp Lethality Test (BST) dari daun beberapa species benalu yang secara tradisional digunakan untuk mengobati tumor di Indonesia

ABSTRACT Background : Traditionally some benalu species were used to treat tumour in Indonesia. Some research report indicated the present of anti tumour activity, although its toxicity study by BST was not performed yet. It is necessary to observe the toxic level of the benalu leaves and whether they are safe to be consumed. Objectives : To determine the toxic level of species of benalu leaves (Dendrophtoe pentandra L. Miq. (1), Macrosolen tetragonus (bL) Miq. (2 and 7), Helixanthera parasitica Lour. (3), Dendrophtoe falcate (Lf.) Ettings (4), Dendrophtoe constricta Dans. (5), Macrosolen cochinchinensis (Lour). Tiegh. (6)) Methods : The leaves of benalu were separately extracted with chloroform, followed by methanol. The chloform and methanol extracts toxicity level were screened using Brine Shrimp Lethality Test (BST) at the dose of 1500, 1000 and 500 ug/mI. The level of the toxicity was determined by counting the death percentage of the Artemia saline larvae after 24 hours of adding the extract at the dose indicated. The extract was considered toxic when 100% A. saline larvae was killed at the dose < 1000 ug/ml, and the extract was mildl toxic when 100% death percentage was observed at the dose of 1500 ug/ml. Results : Practically all the chloroform extracts were non toxic since they were not able to kill 100% A. saline larvae at the dose of 1000 ug/mI. Although when the doses were raised to 1500 ug/mI, the 100% death percentage was still not obtained. Similarly, the methanol extracts were also non toxic at the dose of 1000 ug/ml. However, M. cochinchinensis (6) and M. tetragonus (7) were considered to be mild) toxic as their methanol extracts were able to kill 100% A. saline larvae when the dose was raised to 1500 ug/mI. Conclusion: The different test result (at 1500 ug/mI) shown by two methanol extracts of similar species M. tetragonus {2b (killed 52%) & 7b (100%)} should give an idea that the benalu are practically non toxic and save to be consumed for tumor treatment. Key words : toxicity screening, brine shrimp lethality test (bst) - benalu leaves - benalu hosts - anti tumor

BIOACTIVITY SCREENING OF SPONGES COLLECTED FROM BUNAKEN, MENADO BY Brine Shrimp Lethality Test AGAINST Artemia salina Leach

Indonesia is one of the countries rich of natural resources. Sponges are marine invertebrates widely found in Indonesia and known to have various bioactive compounds with unique chemical structures. This study was aimed to screen potentially bioactive extracts of sponges collected from Bunaken and identify on their toxicity level against larva Artemia salina Leach. Samples were macerated using acetone followed by partition using chloroform and methanol. The extracts were prepared at the concentration of 100, 250, 500 and 1000 mg/ml and their toxicity was tested using Brine shrimp Lethality Test. LC50 (mg/ml) of the extracts was calculated using probit analysis. From the 4 extracts tested, all of them showed toxicity to larva Artemia. Chloroform extract of MD-02 was found to be the most toxic with the LC50 of 48.15 mg/ml and considered as a potential candidate for new drug research. Further characterization is still needed for the development in the future. This active sponge was then identified as Petrosia sp.Key words: sponges, Bunaken, bioactivity, Artemia salina Leac

Antiplasmodial activity of two fractions obtained from n-hexane extract of Garcinia parvifolia Miq stem bark

Antiplasmodial activity of fractions A and B obtained from n-hexane extract of stem bark of Garcinia parvifolia have been evaluated. The in vitro antiplasmodial activity was investigated on two strains of Plasmodium falciparum, FCR-3 a chloroquine resistant and D10, a chloroquine sensitive strains and their antiplasmodial activity was expressed by the concentration inhibiting 50.% of the parasite growth (IC50). The results showed that the fractions A and B were active against P. falciparum with the IC50 values of 2,79 ± 0,10 μg/mL and 12,30 ± 1,21.μg/mL on FCR-3 strain and 1,52 ± 0,24.μg/mL and 4,66 ± 1,24 μg/mL on D10 strain. Identification of active constituents in the both fractions showed the existence of triterpenoide, steroide and flavonoide compounds.Key words: Antiplasmodial activity , Garcinia parvifolia, active constituents

Isolation of aphrodisiac active fraction from sanrego bark (Lunasia amara Blanco)

Sanrego (Lunasia amara Blanco) bark is traditionally utilized as an aphrodisiac although scientifically has yet been proven. Therefore, this study is aimed to prove aphrodisiac activity, determine the compounds and the dose of the active fraction. Initially sanrego bark (600 g) was extracted in a Soxhlet apparatus with methanol (MeOH) to give MeOH extract (A, 80 g). The MeOH extract was fractionated by Ethylacetate (EtOAc) to give EtOAC soluble (B, relatively non-polar, 15.23 g) and EtOAc insoluble (C, relatively polar, 50.20 g). The aphrodisiac test was performed in male Wistar rats that were divided into 6 groups (5 rats each) [I, treated with Na-CMC 0.5%, 50 mg/kg BW; II, yohimbine, 5 mg/kg BW; III, distilled water , 2 ml/200 g BW; IV, extract A; V, fraction B; VI, fraction C], and the doses given to groups IV-VI were similar (10, 50, 100, and 200 mg/kg BW). The male’s behaviors to female rats ( introduction, climbing and coitus ) were recorded and analysed at p= 0.5. Determination of the active compounds were performed by thin layer chromatography (TLC) using various detection reagents. The result indicated that the highest aphrodisiac effect was demonstrated by fraction B, followed by extract A and fraction C . Fraction B demonstrated introduction (84.2%), climbing (84.9%) and coitus (85.2%). TLC profile suggested that fraction B contain alkaloids and terpenoids as the main components.Keywords: Sanrego bark, Lunasia amara, Aphrodisiac, Active fraction

Skrining senyawa bioaktif dari beberapa ekstrak tumbuhan asal kawasan hutan Kalimantan Tengah dan isolasi senyawa bioaktif Fibraurea chloroleuca Miers = Bioactive compound screening from ...

ABSTRAK Indonesia is known for its largest biodiversity on the world. However, up to now they have not been studied completely for their utilization and development especially as drug resources. Bioactive screening on 14 extracts (chloroform and methanol extracts of medicinal plants) using Brine Shrimp Lethality (BST, using larvae Artemia salina) assay (500 and 1000 (iy/ml) gave 17 extracts potential (100% larvae death) to be developed as bioactive compounds resources. Further screening of these 17 extracts at a lower dose (100 (iy/ml), 10 extracts (100% larvae death) were obtained. One of those extracts is chloroform extracts of # 03 bfar 029 identified as Fibraurea chloroleuca Miers. Bioassay (BST) guided isolation of F. chloroleuca extract, the most toxic isolate (bl) was obtained (LC^ 4.5 ny/ml). The isolate (bl) at the highest tested dose (500 ny/ml) inhibited 20,08% of the growth of HeLa cell in vitro. Isolate (bl) appears as orange powder, having melting point at 182.3-183.0°C and X^ (CHCL3) at241 and 345 nm. Based onTLC pictured visualized by Dragendorff, bl is confirmed as an alkaloid. Mass spectrum (EIMS) data of bl showed the highest mass peak at m/z 351 (75%). The odd mass number strengthens the conclusions that bl is an alkaloid having odd number of -N- atom. The TR spectrum (KBr) indicates the present -OH, =CH, -CH, and C=C-C=O, secondary amine functional groups, in addition aromatic ring absorption band is also observed. Keywords: Fibraurea chloroleuca Miers., bioactive, BST, HeLa cel