The main aim of this study was to isolate and characterise antifungal and antibacterial compounds from leaf extracts of Combretum molle which belonging to the Combretaceae family. C. molle is one of the commonly used medicinal plants in southern Africa for numerous ailments. Three animal fungal pathogens, namely, Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus and five plant fungal pathogens, namely, Aspergillus niger, Aspergillus parasiticus, Fusarium oxysporum, Penicillium janthinellum, Rhizoctonia solani and four nosocomial bacteria Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa were used as test microorganisms for bioactive compounds in leaf extracts of C.molle. Experiments for phytochemical analysis were done using different C. molle leaf extracts which were made using acetone, methanol, ethanol, ethyl acetate, chloroform, butanol and hexane as extractants. Thin Layer Chromatography (TLC) fingerprints of different leaf extracts were developed in three mobile phase systems, EMW, CEF and BEA and detected with vanillin-sulphuric acid spraying agent. The different extracts of C. molle showed the presence of many different compounds with distinct retardation factors (Rf), separated according to their polarities. Bioautography was carried out to determine the number of active compounds and their Rf values. The TLC plates were developed in three mobile systems, each sprayed with either fungal or bacterial strains. In BEA bioautograms of A. fumigatus, clear zones of inhibition were observed at Rf values of 0.12, 0.23, and 0.40. In EMW bioautogram of C. albicans, clear zones of inhibition were observed at Rf value of 0.73, 0.81, 0.87. C. neoformans had weak growth inhibition. Most of the fungal and bacterial strains tested in the bioautography displayed susceptibility to the active compounds, with P. janthinellum and P. aeruginosa showing exceptional sensitivity. The minimum inhibitory concentrations (MIC) values ranged from 0.02 to 2.5 mg/ml against the tested pathogens. The acetone and ethyl acetate extracts had the best inhibitory activity against P. janthinellum with an MIC value of 0.02 mg/ml. The acetone extract of C. molle gave the highest total activity (775 ml/g) against P. janthinellum. C. albicans was the most resistant pathogen with an average MIC value of 0.56 mg/ml compared with the other tested strains. Extracts were active against both Gram-positive and Gram-negative strains. P. aeruginosa extracts had the highest average MIC value (0.24 mg/ml) among the tested bacterial strains. In general, there was good overall inhibitory activity by different extracts of C. molle. Bioassay-guided fractionation of DCM extract of the leaves of C. molle yielded 32 fractions. Further fractionation led to the isolation of five compounds (C1, C2, C3, C4 and C5). Compound C1 was selected for structure elucidation due a larger quantity isolated and higher antimicrobial activity compared with the other isolated compounds. Nuclear magnetic resonance (NMR) spectroscopy and mass spectroscopy (MS) was used to show that compound C1 was taraxerol, belonging to the taraxerane group. Antimicrobial activity of the isolated compound against P. janthinellum had an MIC value of 0.08 ug/ml. Although the compound taraxerol have been discovered in other plant species, it is reported for the first time from C. molle in the study. The results illustrate that crude extracts and compound taraxerol from C. molle can be used as either an antibacterial or antifungal, and warrants further investigation. / Dissertation (MSc)--University of Pretoria, 2017. / Paraclinical Sciences / MSc / Unrestricted

M. Tech. (Department of Biotechnology, Faculty of Applied and Computer Sciences) Vaal University of Technology. / Chemical precipitation method is the most widely used of all methods for preparing good quality semiconductor nanoparticles. Several conditions are optimized for producing the desired size and shape of particles. The parameters such as capping molecule, precursor concentration, time and temperature were investigated using the colloidal hot injection method. The effect of capping agent was the first parameter investigated in the synthesis of copper selenide, copper sulphide and copper oxide nanoparticles. The capping agents of interest in this study were oleylamine (OLA) and trioctylphosphine (TOP), due to their ability to act as reducing agents, surfactant, solvent and enhancement of colloidal stabilization. The use of oleylamine and trioctylphosphine were carried out at 220 °C for 30 minutes. The optical and structural properties of the yielded nanoparticles were characterize using UV/Vis spectroscopy, TEM and XRD and showed dependence on the type capping interactions from the two agents. Nanoparticles synthesized using TOP produced two phases whereas a single phase was observed from OLA as confirmed by XRD. OLA produced bigger particle sizes compared to TOP but with a wider variety of shapes. The wide variety of particle structures of OLA capped nanoparticles was advantageous since different types of bacteria were targeted in this work. Therefore, other synthetic parameters were investigated using OLA as both solvent and capping molecule. Precursor concentration ratio showed bigger effect in the size, and shape of the yielded nanoparticles. For copper selenide and copper sulphide (Cu: Se/ S), 1:1 concentration ratio gave the best optical and structural properties while copper oxide (CuO) nanoparticles demonstrated its best optical and structural properties in 2:1 ratio (Cu: O). Nonetheless, 1:1 precursor concentration ratio was used to optimise other parameters. Since reaction time has a profound effect on the nanocrystals size and shapes, the effect of reaction time in OLA was also investigated. The reaction time showed no effect on the phase composition of the synthesized copper sulphide, copper oxide and copper selenide nanoparticles. Reaction time of 30 minutes gave the best optical (the shape of the absorption band edge and emission maxima values) and structural (size distribution of particles) properties for CuSe and CuS compared to other reaction times (15 min, 45 and 60 min). 15 min reaction time gave the best optical and structural properties for copper oxide but nonetheless, 30 min was used as the optimum reaction time for further optimization. Temperature showed an effect in size, shape and the stoichiometry of the reaction. These effects were confirmed by the optical and structural properties of the synthesized nanoparticles. XRD patterns revealed some differences with the temperature change, indicating an effect on the phase composition of CuS and CuO but not on CuSe nanoparticles. CuSe and CuS nanoparticles synthesized at 220 °C gave the ideal optical and morphological features compared to other temperatures that were selected (160 ºC, 190 ºC and 240 ºC). Nonetheless, CuO revealed its best optical and structural properties at 160 ºC. 220 ºC was deduced to be the optimum temperature for the synthesis of these three materials under the synthetic conditions. The optimum parameter (220 ºC, 30 min and 1:1 ratio) were used to synthesize the three copper chalcogenides which were then tested against Gram-negative (E. coli and P. aeruginosa), Gram-positive (S. aureus and E. faecalis), and fungi (C. albicans). The plant species, Combretum molle and Acacia mearnsii were phytochemical screened for the presence of active organic compounds and the content of total phenols, flavonoids and antioxidants using different solvents. Both C. molle and A. mearnsii revealed the highest phenolic content in acetone extracts. C. molle revealed its highest flavonoid content in methanol extract and its highest free radical scavenging activity in acetone extract. Acetone extracts demonstrated the highest flavonoid content as well as the highest free radical scavenging activity of A. meansii. The solubility of copper chalcogenides and plant extract was tested in four different solvents and the solvent that demonstrated highest solubility was used for the coordination of the plant extract and copper chalcogenides. The plant extract coordinated nanoparticles were tested for their antibacterial and antifungal activity. Their results were compared to those of the active ingredient in their respective solvents from the medicinal plants as well as those of copper chalcogenides nanoparticles without plant extracts using diffusion disk and MICs methods. The synthesized nanoparticles showed better performance than plant extracts with copper oxide performing the best, followed by copper selenide and lastly by copper sulfide. The performance of plants extracts highly dependent on the solvent of extract with acetone showing the best performance for both C. molle and A. Mearnsii followed by ethanol. The addition of active ingredients from C. molle and A. mearnsii to the synthesized nanoparticles did not enhance the performance of these nanoparticles.

M. Tech. (Department of Chemistry, Faculty of Applied and Computer Sciences), Vaal University of Technology / The present study includes the use of a green synthetic method to prepare copper and silver nanoparticles using chitosan, aqueous extracts of Camellia sinensis, Combretum molle and Melia azedarach linn leaves. This study aims to investigate the influence of capping and precursor concentration on the properties of silver nanoparticles with emphasis on the medicinal plants chosen. The effect of capping agent on the properties of copper nanoparticles is also investigated. The phytochemical properties of plant extracts and the antimicrobial activity of the synthesized particles were also studied; this was achieved by using microdilution bioassay. Decoction method was used to extract secondary metabolites from plant leaves. Preliminary phytochemical screening carried out on the aqueous extracts of the plant leaves showed the presence of tannins, proteins, flavonoids, phenols, and carbohydrates. The total phenolic and flavonoids content of the aqueous extract was determined using spectroscopic methods. The highest phenolic content was found in the aqueous extract of Combretum molle (135 mg/g), and the highest flavonoid content was found in the aqueous extract of Camellia sinensis (0.4 mg/g). Characterization was done by a combination of spectroscopic, microscopy and XRD techniques. Both the size and shape of the synthesized silver nanoparticles were dependent on the identity of the capping molecule, precursor and capping agent concentration as depicted from their TEM and XRD results. Silver nanoparticles were found to be predominantly spherical. The capping agent concentration was also found to influence the degree of agglomeration, with an increase in capping agent concentration giving lesser agglomeration. FTIR spectral analysis showed that silver nanoparticles interact with bioactive compounds found in the plants through the hydroxyl functional group. Other shapes including diamond were observed for the effect of precursor concentration. The XRD micrographs revealed a face-centered cubic geometry and the phase remained the same with an increase in precursor concentration. The synthesized silver nanoparticles were all blue shifted compared to the bulk material. The TEM results revealed that copper nanoparticles with different sizes and shapes were successfully synthesized. All the prepared copper and silver nanoparticles showed satisfactory antifungal and antibacterial activity against Candida albicans, Cryptococcus neoformans, Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumonia and Pseudomonas aeruginosa. The capping molecules used in this study also showed some antibacterial and antifungal activity against the selected strains. However nanoparticles performed better than these capping molecules. Both silver and copper nanoparticles were found to be more active against gram-negative bacteria compared to gram-positive bacteria. Amongst all the prepared silver nanoparticles Combretum molle capped nanoparticles were found to be the most active nanoparticles. Also with copper nanoparticles, it was found that Combretum molle capped nanoparticles were the most active nanoparticles. Between the two metal nanoparticles, silver nanoparticles showed high antibacterial and antifungal activity compared to copper nanoparticles. The antioxidant activity of silver nanoparticles was assessed using 2.2-diphenyl-1-picrylhydrazyl. Silver nanoparticles were found to have some antioxidant activity. However, the capping molecules were found to be more active than the synthesized nanoparticles. This observation is attributed to the presence of some bioactive compounds in the plant extracts.