3. Electronic Theses and Dissertations (ETDs) - All submissions
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Item Indigenous Salvia species : an investigation of their pharmacological activities and phytochemistry(2008-09-26T07:03:36Z) Kamatou, Guy Paulin PoungoueThe genus Salvia belongs to the family Lamiaceae and encompasses 900 species worldwide of which 26 are found in southern Africa and many of them are used in local traditional medicine. However, the phytochemistry and pharmacological activities of the South African species have not been extensively investigated. The leaf trichome morphology that may be used to distinguish species was investigated with the scanning electron and light microscopy. Both glandular (capitate or peltate) and non-glandular trichomes were identified in all species. The essential oils were isolated by hydro-distillation and analysed by GC and GC-MS methods. The oil yield was relatively low and ranged from 0.004 (S. radula) to 0.50% (S. muirii) (w/w). Major components identified include α-pinene, 1,8-cineole, linalool, limonene, myrcene, β-caryophyllene, spathulenol, β-caryophyllene oxide, viridiflorol, δ-3- carene and α-bisabolol. High performance liquid chromatography analysis was used to identify phenolic compounds in 17 solvent extracts. Betulafolientriol oxide was detected in all species. Rosmarinic acid was only absent in S. verbenaca, while S. garipensis and S. radula were the only species which lacked oleanolic acid/ursolic acid. Various in vitro biological activities were investigated. Nearly all the solvent extracts displayed anti-oxidant activity with IC50 values ranging from 1.61 to 74.50 μg/ml using the DPPH· radical, while the IC50 values ranged from 11.88 to 69.26 μg/ml with the ABTS·+ radical. The solvent extract of S. schlechteri was three times more active than vitamin C. Total phenolic content based on gallic acid equivalents (GAE) revealed the presence of total soluble phenolics in the extract at 45 to 211 mg of GAE dry sample. Almost all the essential oils exhibited promising anti-inflammatory activity (5-lipoxygenase assay) with IC50 values ranging from 22.81 to 77.32 μg/ml. The antimalarial activity was determined using [3H]-hypoxanthine method on the Plasmodium falciparum (FCR-3) strain. The IC50 values of the essential oils ranged from 1.20 to 13.50 μg/ml and were low compared to the solvent extracts (IC50 values ranging from 3.91 to 26.01 μg/ml). Betulafolientriol oxide and salvigenin isolated from S. radula inhibited the growth of malaria parasites with IC50 values of 4.95 and 24.60 μg/ml, respectively. With the exception of S. radula, all the solvent extracts displayed moderate to good activity against Staphylococcus aureus, vii Bacillus cereus, Klebsiella pneumoniae, Escherichia coli and Mycobacterium tuberculosis with the MIC values ranging from 0.03 to 8.00 mg/ml. Four compounds, namely carnosol, 7-O-methylepirosmanol, oleanolic acid and its isomer ursolic acid were isolated from S. chamelaeagnea as the active principles against S. aureus. The solvent extracts of Salvia species were tested for in vitro anticancer activity against human breast adenocarcinoma (MCF-7), colon adenocarcinoma (HT-29) and glioblastoma (SF-268) using the sulforhodamine B assay. The extracts inhibited cell proliferation of all three cell lines to varying degrees, with the IC50 values ranging between 9.69 and 43.65 μg/ml and 8.72 and 59.12 μg/ml against the MCF-7 and SF-268 cell lines, respectively. The IC50 values against the HT-29 cell line ranged from 17.05 to 57.00 μg/ml. The in vitro toxicity profile of 28 samples (17 solvent extracts and 11 essential oils) was evaluated on human kidney epithelial cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5 dimethyl tetrazolium bromide method. The samples displayed some degree of toxicity with IC50 values ranging from 1.79 to 22.9 μg/ml for the essential oils and from 12.12 to 53.34 μg/ml for the solvent extracts. The essential oil composition of S. africana-caerulea, S. africana-lutea and S. lanceolata, collected at the same locality throughout the 2004/2005 growing season, was compared in terms of essential oil yields, chemical composition and biological activities. Mostly quantitative, rather than qualitative variation was observed. Major seasonal fluctuations of certain essential oil compounds were observed in all three species. Variations in biological activities of the solvent extracts over seasons were noted. The biological activities of the solvent extracts of three Salvia species (Salvia africanacaerulea, S. africana-lutea and S. lanceolata) were evaluated in the presence and absence of essential oils. The solvent extract of S. africana-caerulea without essential oil exhibited the best activity against Gram-positive bacteria (MIC value: 0.1 mg/ml), while the solvent extract containing essential oil of S. africana-lutea was the most active against Gramnegative bacteria. The toxicity profile of all three species was significantly higher (P < 0.05) with the solvent extracts containing essential oils. The in vitro biological activities add scientific support to the use of Salvia species in traditional medicine.Item The phytochemistry and microbial activity of selected indigenous Helichrysum species(2008-06-10T07:01:25Z) Reddy, DakshinaABSTRACT Helichrysum (Asteraceae) is a large genus consisting of approximately 500 species of which 245 taxa are indigenous to southern Africa. Many Helichrysum species are widely used by the indigenous population to treat various ailments, including coughs, colds, fever, infections, headache, menstrual pain and as a treatment for wounds. Medicinal uses are often not species-specific but often depend on the local availability. Guided by the traditional use and the lack of scientific information, nine species of Helichrysum were selected for this study. The essential oils were obtained through hydrodistillation and methanol and acetone extracts of the plant material were prepared. The essential oil composition was determined using GC-MS. The oil profiles were mostly dominated by the presence of monoterpenes such as a-pinene, 1,8-cineole and p-cymene. Monoterpenes were largely absent in the essential oil of H. felinum, but this oil was rich in sesquiterpenes with high yields of b-caryophyllene. The antimicrobial activity of the essential oils and plant extracts were of interest due to their traditional use as an antiseptic. The antimicrobial activity of the essential oils and extracts was determined by disc diffusion assays and, following this, the most active species were further investigated using the minimum inhibition concentration (MIC) assay. Helichrysum dasyanthum displayed the best activity against B. cereus (MIC = 16 mg/ml) and was the only extract that exhibited activity against all three fungal strains tested (C. neoformans, 1 mm; C. albicans, 3 mm; and A. alternata, 2 mm). The essential oil of H. petiolare and H. felinum exhibited the most pronounced activity against the fungal strains in the disc diffusion assay (C. albicans, 2mm).Item The biological activity and phytochemistry of selected Hermannia species(2006-10-31T11:57:46Z) Essop, Ayesha BibiTraditional medicines form a significant part of the lives of many people around the world and in South Africa almost 60 % of people consult traditional healers in addition to the modern medical services available. Plants form a significant part of traditional healing and hence, selected species of a traditionally used plant genus, Hermannia, were chosen for biological and chemical investigation to determine a scientific basis for the traditional use of these plants. A phytochemical investigation was carried out, firstly using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) and then isolation and identification of compounds from various Hermannia species. TLC analysis indicated significant similarities between the various species with only H. saccifera displaying chemical anomalies. This was further corroborated by the HPLC analysis although very conservative profiles were produced. Isolation of compounds from H. saccifera yielded a novel labdane compound, E-17, 19-diacetoxy - 15 - hydroxylabda - 7,13 - diene, as well as two flavones, 5,8- dihydroxy-6,7,4’- trimethoxyflavone and cirsimaritin which have previously been isolated. In addition, two commonly found compounds, lupeol and β- sitosterol were isolated from H. cuneifolia and H. salviifolia respectively. This is the first report on the isolation and identification of all five compounds from Hermannia species. Antimicrobial activity was assessed using two methods i.e. minimum inhibitory concentrations as well as the death kinetics assay. Minimum inhibitory concentrations were determined using four Gram-positive and two Gram-negative bacteria as well as two yeasts. All species investigated indicated antimicrobial activity with H. saccifera showing good activity against S. aureus and B. cereus. E-17, 19-diacetoxy - 15 - hydroxylabda - 7,13 - diene isolated from H. saccifera indicated activity (MIC = 23.6 μg/ml against S.aureus) although the activity was less than that of the crude extract (MIC = 19.5 μg/ml), thus, demonstrating that there are a number of compound contributing to the promising activity of the crude extract. This was further corroborated by the bioautograms developed of the H. saccifera extract. Time-kill studies on H. saccifera against S. aureus indicated that at concentrations of 0.1, 0.25 and 0.5 % bacteriostatic activity was observed while at 0.75% the extract achieved complete bactericidal activity after 240min. Free radical scavenging activity was assessed using the 2,2-diphenyl-1-picrylhydrazy (DPPH) and 2,2′-azino-bis(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS) assays. Ten of the twelve species indicated good activity with H. cuneifolia demonstrating the most promising activity (IC50 = 10.26 μg/ml for DPPH and 10.32 μg/ml for ABTS). Two of the isolated compound, 5,8- dihydroxy-6,7,4’- trimethoxyflavone and cirsimaritin displayed insignificant activity. The 5-lipoxygenase assay was used to assess the anti-inflammatory activity of Hermannia species. All species exhibited intermediate activity with the exception of H. cuneifolia (IC50 = 15.32 μg/ml). In addition, four isolated compounds, 5,8- dihydroxy-6,7,4’- trimethoxyflavone, cirsimaritin, lupeol and β-sitosterol showed moderate inhibition of the enzyme indicating that while these compounds do contribute to the activity of the extracts they are not individually responsible for any significant activity. Antimalarial activity was assessed using the titrated hypoxanthine incorporation assay while toxicity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cell proliferation assay. Only three species indicated any good antimalarial activity i.e. H. saccifera, H. muricata and mostly H. trifurca (IC50 = 25.30, 28.17 and 18.80 μg/ml respectively). However, the activity of H. saccifera and H. trifurca are probably due to a general cytotoxicity as these species exhibited a low safety index. All other species appear safe for use. Several Hermannia species have indicated in vitro biological activity in a number of assays which is related to their use in traditional medicines to treat a number of disease states. Hence, a scientific basis, albeit in vitro, has been established for the use Hermannia species in traditional healing.