IPRC - 2015
Permanent URI for this collectionhttp://repository.kln.ac.lk/handle/123456789/156
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Item New Bioactive poliketide from endolichenic fungus, Curvularia trifolii, inhabiting the lichen, Usnea sp. collected from Haggala Botanical Garden(Faculty of Graduate Studies, University of Kelaniya, 2015) Samanthi, K.A.U.; Wickramaarachchi, S.; Wijerathne, E.M.K.; Paranagama, P.A.Endolichenic fungi (ELF), which live in healthy tissues of lichens, are one of the promising producers of structurally diverse novel bioactive secondary metabolites. It has been reported that endolichenic fungi are rich sources for the discovery of new compounds with significant medicinal values and this was demonstrated by isolation and identification of new anticancer, antifungal, antibacterial and antioxidant agents from endolichenic fungal extracts. In Sri Lanka, research group of University of Kelaniya initiated a research program on ELF and have isolated few interesting bioactive poliketides from ELF of three lichen species available in Sri Lanka. They have shown great potential to be a source for novel bioactive natural products. The objectives of this study were isolation and identification of bioactive compounds of the endolichenic fungus, Curvularia trifolii isolated from the lichen Usnea species. In a continuation of our chemical investigations in search of secondary metabolites of endolichenic fungi with medicinal value led to the isolation of few bioactive compounds. In this present study, C. trifolii was grown in large scale and incubated at room temperature for 10 days. The metabolites were extracted into ethyl acetate (EtOAc). Antibacterial, anticancer and insecticidal activities of the EtOAc extract were evaluated using in-vitro bioassays. The EtOAc extract was subjected to a silica gel column chromatography resulting, one major compound with high bioactivities. The isolated pure white crystalline compound was identified as 1,14-dihydroxy-6-methyl-6,7,8,9,10,10,14,14 -octahydro-1H-benzo[f][1] oxacyclododecin-4(13H)-one using 1H, 13C NMR, 2D NMR and MS data. The compound was found to possess antibacterial activity against three test pathogens, anticancer activity at (NCIH-460), (MCF-7), (SF-268), (PC3M), and (MDA-MB-231) cell lines and insecticidal activity.Item Isolation of bioactive secondary metabolites from the endolichenic fungi, Neosartorya sp. inhabiting the lichen Parmotrema sp. in Sri Lanka(Faculty of Graduate Studies, University of Kelaniya, 2015) Manthrirathna, M.A.T.P.; Kandiah, R.; Gunasekara, D.S.; Paranagama, P.A.Natural products are a potential source of novel pharmaceutical agents. Therefore, isolation and identification of bioactive compounds from organisms adapted to various biotopes and unraveling their bioactivities in search for new pharmacophores has a mounting interest. Fungi are known to be prominent producers of useful metabolites. Endolichenic fungi (ELF) that occur asymptomatically within the lichen thalli are one of the ecological groups of fungi. ELF in Sri Lanka remain almost unexplored as a source of useful bioactive compounds. The objective of this study is to isolate bioactive secondary metabolites from ELF Neosartorya sp. isolated from Parmotrema sp. that occur in Hakgala Botanical Garden. ELF Neosartorya sp. was cultivated on 48 PDA plates and incubated at room temperature. Secondary metabolites were extracted into ethyl acetate from 9 days old cultures. Antibacterial activity of the crude extract was evaluated against Bacillus subtilis (BS) and Staphylococcus aureus (SA) using agar well diffusion method. Standard antibiotic Azithromycin was used as the positive control and Dimethyl sulfoxide as the negative control. Since the crude extract showed antibacterial activity against both BS and SA, it was partitioned with hexane, chloroform (CHCl3) and aqueous methanol. All three fractions showed activity against SA, with the CHCl3 fraction having higher activity compared to the other two fractions. Chloroform and methanol fractions showed significant activity against BS, while CHCl3 fraction showed activity comparable with Azithromycin. Chloroform fraction of Neosartorya sp. was further fractionated using bio-assay guided fractionation (silica gel column chromatography). Pure compounds were isolated using preparative TLC. One major pure compound was isolated from CHCl3 fraction and the characterization still in progress.Item Investigating the bioactive compounds of endolichenic fungus Hypoxylon anthochroum isolated from the lichen, Pseudocyphellaria sp.(Faculty of Graduate Studies, University of Kelaniya, 2015) Antony, K.N.L.; Samanthi, K.A.U.; Paranagama, P.A.It has been reported that 11 endolichenic fungal strains were isolated from lichen, Pseudocyphellaria sp. Endolichenic fungi live asymptomatically within the lichen thallus and known as a new source useful of bioactive secondary metabolites. Secondary metabolites in endolichenic fungi show distinct bioactivities including antimicrobial, anti-inflammatory, analgesic, antioxidant and cytotoxic activities. These bioactive compounds have potential importance in development of pharmaceutical drugs, nutraceuticals and agrochemicals. The present study was carried out to investigate antioxidant activities of secondary metabolites produced from Hypoxylon anthochroum inhabiting the lichen, Pseudocyphellaria sp. In this research, H. anthochroum was cultured in 50 large petridishes containing potato dextrose agar (PDA) medium and incubated at room temperature for seven days. After the incubation period, secondary metabolites were extracted into ethyl acetate in order to obtain crude extract. The crude extract was partitioned using solvents hexane, chloroform and 60% aqueous methanol. Antioxidant activities of three fractions were evaluated using micro plate DPPH assay. Since chloroform fraction showed the highest antioxidant activity (81.96 % Inhibition at 800 μg/mL) it was further subjected to silica gel column chromatography and preparative thin layer chromatography to isolate bioactive compounds. One UV active pure compound which has antioxidant activity (43.21 % Inhibition at 800 μg/mL) was isolated from chloroform fraction and its structure elucidation will be carried out using 1H NMR, 13C NMR, and 2D NMR spectral data.Item Evaluation of Essential Oil content and composition of Ocimum Species in Five Districts of Sri Lanka(Faculty of Graduate Studies, University of Kelaniya, 2015) Aththanayaka, A.M.L.; Dharmadasa, R.M.; Paranagama, P.A.The genus Ocimum is a member of the Lamiaceae family which contains about 150 species of herbs and shrubs. Majority of species of genus Ocimum are essential oil bearing plants. Several species belonging to genus Ocimum are distributed in Sri Lanka. In the present study attempts were made to investigate the essential oils of populations distributed in 5 districts of Sri Lanka. Survey was conducted Anuradhapura, Matale, Gampaha, Kurunegala and Puttalam districts and 9 different samples (APMI 3, APMI 4, MLLD 2, MLLD 5, MLLD 6, KGYP 2, KGYP 5, GPLG 5, and PTWW 2) were collected, Species were authenticated by comparing with National Herbarium. These samples were air dried for 3 days and cut into small pieces (2cm) and hydro distilled using Clevenger apparatus. Essential oil composition was determined using GC-MS analysis. Cluster analysis for essential oil composition was carried out by using SPSS version 14. Oil content varied from 0.4% to 1.6% depending on the locality and the population. The highest oil content was reported from Loluwagoda (GPLG 5) in Gampaha district while the lowest was from Yaggapitiye, in Kurunegala district. Eugenol content varied from 1.45% (APMI 4) to 86.33% (GPLG 5). Methyl eugenol, Camphor, was only found in APMI 4 (85.64%) and APMI 3 (37.99%) respectively as the main component. Germacrene-d and Caryophyllene are common compounds of all the populations except APMI 3. Ocimine was found in 6 species except APMI 3, APMI 4, and MLLD 6. Thirty six compounds were identified from nine species. All populations were clustered into two on the basis of availability of different compounds using hierarchical cluster analysis. Cluster 1 consisted of 3 populations (MLLD 5, GPLG 5, KGYP 5) while cluster 2 consisted of 6 populations ( MLLD 2, KGYP 2, PTWW 2, APMI 3, APMI 4, and MLLD 6). Information generated through the present study could be of vital importance for chemotaxonomy of Ocimum populations.Item Bioactive compounds and in-vitro antioxidant potential of mango (Mangifera indica L.) flesh, peel and seed(Faculty of Graduate Studies, University of Kelaniya, 2015) Thiripuranathar, K.G.; Navaratne, A.N.; Paranagama, P.A.Natural antioxidant compounds from flesh, peel and seed of fruits have gained increasing attention in the scientific community. Synthetic antioxidants cause the side effects such as cancer and cardiovascular diseases. The antioxidants from natural sources are the only alternative to synthetic antioxidant. The main objective of this study was to evaluate the potential efficacy of mango peel and seed kernel as an antioxidant.In the present study three mango varieties ‗Willard‘, ‗Karuthacolomban‘ and ‗Vellaicolomban‘ were collected from Jaffna district during April to July, 2015. Ethyl acetate extracts of flesh, peel and seed kernel of three mango varieties were studied for the total phenolic content (TPC) and total flavonoid content (TFC) using Folin-Ciocalteu method and aluminium chloride colorimetric assay respectively. A study of in vitro antioxidant activities were also studied using different assays, which include such DPPH radical scavenging assay, Ferric ion reducing power assay (FRAP), Nitric oxide scavenging assay (NO.) and ABTS radical scavenging assay(ABTS.+).The results revealed that peel of Willard showed the highest TPC (275.61±5.24 mg GAE /g extract) and flesh of Vellaicolomban showed the highest TFC (479.8015.30 mg QE/g extract). The DPPH radical scavenging activity results revealed that the ethyl acetate extract of Karuthacolomban seed kernel (IC50-7.7±0.4 μg/mL) showed a higher radical scavenging activity than that of the standard, ascorbic acid (IC50- 11.0±0.4μg/mL). Peel obtained from Willard and seed kernel obtained from Karuthacolomban showed the highest antioxidant activity in FRAP assay. The NO. assay results indicated that seed kernel of Karuthacolomban(IC50-60.6±3.5μg/mL) showed significantly higher scavenging activity than other eight extracts, but it showed lower activity than, ascorbic acid(IC50-22.3±1.6 μg/mL).The ABTS.+ assay indicates that seed kernel of Kruthacolomban (IC50-46.4±1.9 μg/mL) showed higher radical scavenging activity than that of the standard, Trolax (IC50-136.6± 5.2μg/mL). Finding of the present investigation showed that the mango peel and seed kernel contained more polyphenols and exhibited good antioxidant activity than that of flesh.Item Antioxidant active novel secondary metabolite from endolichenic fungus, Penicillium citrinum, inhabiting the lichen, Pamotrema sp. available in Sri Lanka(Faculty of Graduate Studies, University of Kelaniya, 2015) Samanthi, K.A.U.; Wickramarachchi, S.; Wijerathne, E.M.K.; Paranagama, P.A.Item Antifeedant and antifungal activity of heartwood extracts of tea cultivars against Glyptotermes dilatatus Bugnion and Popoff and Fusarium sp.(Faculty of Graduate Studies, University of Kelaniya, 2015) Senanayak, P.D.; Mohotti, K.M.; Paranagama, P.A.The low country live wood termite, Glyptotermes dilatatus is an economically important insect pest of low grown tea. The termite colony initiated by a pair of alates in rotted tea stumps move to the heartwood by making feeding galleries. They disperse wood rot fungi while feeding on heart wood of tea bush. The damage symptoms vary with the type of cultivar. Objective of the present study is to study the chemical constituents in susceptible and resistant cultivars. Extracts of the debarked healthy stems of tea cultivars, TRI 2023 & TRI 4042 (susceptible) and TRI 2027 & TRI 4049 (resistant) were evaluated for the antifeedant and antifungal activities against G. dilatatus and Fusarium sp. which is common in the termite body, termite galleries and rotted tea stems using no choice feeding bioassay and antifungal assay respectively. Extract of TRI 2027 obtained using CHCl3 showed a higher antifeedant activity (P<0.05, LSD) against G. dilatatus than that of TRI 2023, TRI 4042 or TRI 4049. The highest antifungal activity was observed in the extracts of both resistant cultivars (TRI 2027, TRI 4049) than that of susceptible cultivars (P<0.05, Tukeys mean separation test). Fractionation of stem extract of TRI 2027 indicated that caffeine is one of the major bioactive compounds that showing antifeedant and antifungal activities. Antifeedant and antifungal activities were detected at 160 mg/ mL and 0.8 mg/ mL concentrations respectively. Mean caffeine contents of heartwood of resistant and susceptible cultivars were quantified using colorimetric method. Results revealed that the caffeine content of resistant cultivars TRI 2027 and TRI 4049 (0.52±0.26, 0.34± 0.07 mg/g) were greater ( t=3.28,df= 14, P=0.0055) than that of the susceptible cultivars, TRI 2023 and TRI 4042 (0.21±0.08, 0.22±0.01 mg/g) resulting a strong positive correlation between caffeine content and antifeedant and antifungal activities of the extracts of heartwood of tea cultivars. Hence caffeine is proven to be potential biochemical parameter to evaluate resistance of tea cultivars against G. dilatatus and Fusarium sp.Item Impact of a native ‘natural spice mixture’ on selected biomarkers, reduction of excess weight in purposely selected overweight/obese females(Faculty of Graduate Studies, University of Kelaniya, 2015) Manuha, M.I.; Iqbal, N.Z.; Nageeb, B.M.; Paranagama, P.A.The objective of this study is to explore the changes in the body weight and selected biomarkers of overweight/obese women after a four month intervention of native ‗natural spice mixture‘. This study was done at National Ayurveda Teaching Hospital, Borella. Volunteers (129) were participated, and BMI and selected biomarkers (lipid profile) were measured before and after intervention. Chi square test revealed that there is a significant weight reduction after the intervention (Chi Square = 36.74, df = 9, p< 0.05). Although there was no participant with a normal BMI before intervention, 7 % of participants have attained normal BMIs after the intervention. The BMI range 25-29.9 (overweight) have increased by decreasing the ranges 30 – 34.9 (Obesity-1), 35 - 39.9 (Obesity-2) and >=40 (Morbid obesity). Before intervention the percentages of ‗High‘ and ‗Borderline high‘ cholesterol were 17.8 % and 38.8 %, respectively and after the intervention the same were reduced to 7 % and 22.7 %. Before intervention only 43.4 % of the participants were with the normal total cholesterol range, but it increased to 70.3 % after intervention with a significant (p < 0.05) change. Only 6.2 % belonged to the ‗High triglyceride‘ category before the intervention and after the intervention it reduced to 2.2 %. Percentage of ‗Optimal‘ and ‗Near optimal‘ levels of LDLs increased from 17.1 % to 27.6 % and from 28.7 % to 39.4 % after the intervention, respectively. The ‗Borderline‘ and ‗High‘ levels of LDLs were reduced by 5.8 % and 11.6 % after the intervention, respectively. Study shows that 19.4 % of the participants had ‗low HDL‘ before the intervention and it increased to 23.4 % after the intervention. The intervention of ‗natural spice mixture‘ revealed the promising effect on weight reduction and rearranged the dyslipidaemia.