Thesis (MSc.(Biochemistry)) -- University of Limpopo, 2021 / Drug discovery from medicinal plants continues to play an important role in the development of anticancer agents, this is because medicinal plants are reservoirs of bioactive compounds that exert a plethora of pharmacological effects on human beings. This study aimed to analyse the phytochemical constituents of the Momordica cardiospermoides crude acetone and methanol leaf extracts as well as investigate their potential anti-metastatic effects on the MDA-MB-231 breast cancer cell line. Momordica cardiospermoides leaves were extracted with absolute methanol or acetone to produce crude methanol and acetone extracts, respectively. The extracts were then screened and analysed for phytochemicals using thin layer chromatography, qualitative and quantitative phytochemical tests, and their antioxidant activity was determined using the quantitative 2,2-diphenyl-1picrylhydrazyl (DPPH) free radical scavenging activity assay. The fingerprint profiles of the M. cardiospermoides leaf extracts revealed that compounds of the acetone extracts were optimally separated in the nonpolar mobile phase (TAE), whereas those of the methanol extract separated best in the polar mobile phase (EMW), thereby suggesting that the crude acetone and methanol extracts had more non-polar and polar compounds present, respectively. Furthermore, the qualitative phytochemical analysis indicated the presence of various phytochemicals such as flavonoids, steroids, coumarins, and tannins in both plant extracts, however, saponins were found present in the methanol extract and not in the acetone extract. Moreover, quantification of major phytochemicals revealed that the acetone extract had the highest total phenolic content (23.0683 mg GAE/g), total tannin content (22.0442 mg GAE/g) and total flavonoid (32.6933 mg QE/g) content as compared to the methanol extract (14.2349 mg GAE/g, 11.3164 mg GAE/g and 7.692 mg QE/g respectively). The DPPH free radical scavenging activity assay revealed that the extracts exhibited an increase in percentage inhibition/ DPPH scavenging effect, with an increase in extract concentration. The results also revealed that the acetone extract possessed a higher radical scavenging activity as compared to the methanol extract. These results are in correlation with the quantitative analysis of the extracts, as all the major phytochemicals found in higher amounts in the acetone extract have antioxidant properties. The extracts were then assessed in vitro for their cytotoxic effects on MDA MB-231 breast cancer cells and HEK 293 cells using the cell count and viability assay and the results obtained revealed a concentration-dependent decrease in the viability of MDA-MB-231 cells at 24 hours of treatment with either the acetone or methanol extract. Comparatively, treatment of HEK 293 cells with the acetone extract resulted in a significant decrease in the percentage of viable cells, whereas treatment with the methanol extract had no significant effect on the viability of HEK 293 cells, as the percentage of viable cells was maintained at 85–98% at 24 hours of treatment. These results also revealed that the methanol extract is more selective to cancer cells in comparison to the acetone extract, suggesting that the methanol extract is a better antineoplastic candidate. The mode of cell death induced by the methanol or acetone extracts was assessed using the acridine orange and ethidium bromide dual staining assay and the annexin V and dead cell kit. The results from the acridine orange/ethidium bromide dual staining assay showed that both extracts induced nuclei and cellular morphological changes in a concentration-depended manner, at 24 hours of treatment. Moreover, the annexin V and dead assay kit results revealed that the acetone extract induced necrotic cell death, while the methanol extract induced apoptotic cell death. Since the acetone extract was shown to be non-selective towards normal cells and induced necrotic cell death, it was discontinued for further assays. The effect of the methanol extract on MDA-MB-231 cell migration and attachment was determined using the wound healing assay and the adhesion assay. The results revealed that treatment with 150 or 300 µg/ml significantly suppressed MDA-MB-231 cell migration, associated with serpin E1 downregulation and TIMP-1 upregulation, at 24 hours of treatment. Moreover, treatment with the methanol extract also significant inhibited MDA-MB-231 cell adhesion in a concentration-dependent manner, as evident by the decrease in the number of crystal violet stained cells. The effect of the methanol extract on the expression of matrix metalloproteinase-2 and -9 was assessed using western blotting, and the results revealed that the extract significantly downregulated the expression of both MMP-2 and -9, suggesting that the methanol extract has inhibitory effects on MDA-MB-231 cell invasion. The human angiogenesis antibody array kit was then used to determine the effect of the extract on the expression of angiogenesis-related proteins. Treatment with 150 or 300 µg/ml of the extract significantly upregulated the expression levels of tissue inhibitor of metalloproteinases (TIMP) -1 and thrombospondin-1 in a concentration-dependent manner. The results also revealed a significant downregulation in the expression of serpin E1, in a concentration-dependent manner, in comparison to the untreated control. However, the expression of uPA, VEGF, and IGFBP-1, 2 and -3 was upregulated following treatment with 150 and 300 µg/ml of the extract. In conclusion, the current study demonstrated the potential of M. cardiospermoides crude methanol extract as an effective anti-metastatic agent or a source of compounds with anti-metastatic properties / South African Medical Research Council (SAMRC) Research Capacity Development Initiative and National Research Foundation (NRF)

Thesis (M.Sc. Agriculture (Horticulture)) -- University of Limpopo, 2017 / Artemisia annua L. from the family Asteraceae is an annual medicinal plant and has been used to make herbal remedies in Asia for thousands of years. Artemisinin is a sesquiterpene lactone, isolated from aerial parts of Artemisia annua, with the highest concentrations being in flowers and leaves. In addition to potent anti-malarial activity, artemisinin possesses anti-cancer, anti-schistosomiatic, anti-hepatitis B, anti-HIV, anti-leishmanial and herbicidal activities. Low artemisinin production (0.01-2%) from A. annua is a major constraint in commercialisation of the drug for control of malaria. Worldwide, efforts have been underway to improve the concentration of artemisinin using conventional breeding, biochemical, physiological, molecular and hairy-root culture techniques, however all these methods are not economical. Cultural practices like spacing and pruning have limitation in improving artemisinin concentration and these may help in increasing the concentrations of artemisinin. Study was conducted at the experimental farm of the Agricultural Research Council – Vegetable and Ornamental Plants, Roodeplaat Pretoria. The objective of this study was to determine whether spacing, pruning and their interactions would have any effect on the concentrations of artemisinin, growth and yield of A. annua and whether drying methods would have an effect on the concentrations of artemisinin in A. annua. Since there was only one field trial, all sub-objectives were addressed at once (Chapter 3). Fresh seeds of A. annua were obtained from the ARC-VOP gene bank and sown in seedling trays in September 2014. Uniform eight-week-old seedlings were hardened-off, transplanted in November 2014 in 10 cm deep holes and then pruned ten weeks after transplanting. Treatments for Experiment 1, viz., 3 × 4 factorial experiment were laid out in a randomised complete block design, with four replications (n = 48). The two factors of the experiment were (a) spacing [0.5 × 1 m2 (standard: 0.50 m2), 0.5 × 0.7 m2 (small: 0.35 m2) 0.5 × 0.5 m2 (smaller: 0.25 m2) and 0.3 × 0.7 m2 (smallest: 0.21 m2)] and (b) pruning [no pruning (control), removing the apical bud and removing shoots three nodes from the bottom]. The plants were irrigated using overhead sprinklers system for two hours three times per week. Four readings for growth variables (plant height, stem diameter and chlorophyll content) were collected with one week interval. Plants were harvested after 180 days from planting, and leaves, stems and roots were separated weighed and oven dried at 40 ºC for 72 h. In Experiment 2 (drying methods), treatments, namely, 100% sun, 100% shade, 50% shade, freeze and oven drying were arranged in completely randomised design with four replicates (n = 20). The treatments were exposed for a week, to full sunlight, 50% shade-drying under a shade net that allows 50% light penetration, 100% shade under enclosed room at ambient (24-25 ºC) temperature, oven drying for 24 h at 40 ºC, and freeze-drying for three days. Freeze-drying had significant effect on artemisinin concentration of 1.941%. It was followed by oven (1.738%) and 100% shade drying (1.657%) and the lowest artemisinin concentration (1.412%) was obtained from 50% shade drying. The smaller spacing of 0.25 m2 in combination with apical bud removal had a significant effect on artemisinin concentration, producing artemisinin concentration of 0.193%. Spacing had a significant effect on stem diameter, fresh leaf mass and dry leaf mass but had no effect on plant height and chlorophyll content. Pruning had a significant effect on plant height and chlorophyll content and had no effect on stem diameter. The small spacing of 0.35 m2 had the highest fresh and dry leaf mass of 17.99 and 9.62 t/ha. The interaction of spacing and pruning had no significant effect on the growth and yield of A. annua. The results from this study suggested that cultural and processing practices may have direct effects in the concentration of artemisinin, growth and yield of A. annua. The results xiv provided some understanding on how agronomic and processing practices can be used to increase artemisinin content in A. annua and understand the interaction between different agronomic practices and thereby allowing the development of economic methods for A. annua post-harvest handling. Future work should focus on implementing various pruning techniques to trigger stress and indirectly secondary metabolites

No description available.

The anti-inflammatory properties of four medicinal plants were investigated. These plant extracts were subjected to screening for their possible effects as antioxidative, analgesic, and anti-inflammatory agents. In the antioxidant activity, the Plantago lancelota extracts resulted in an IC50 value of 0.4 mg/mL compared to the positive control quecertin with IC50 0.04 mg/mL Plantago lanceolata inhibited COX-2 activity with IC50 values of 0.41 mg/mL. However, the COX-1 inhibition indicated an IC50 of 68.99 mg/mL. The lipoxygenase assay indicated that Plantago lanceolata was the most active plant species with an IC50 value of 4.86 mg/mL compared to the positive control (quecertin) with an IC50<2mg/mL. The nitric oxide assay of the plant extracts indicates a dose-dependent activity of our plant extracts. Likewise the cell viability result indicated a good activity at dose 100 mg/mL. / Life and Consumer Sciences / M. Sc. (Life Sciences)

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 114 leaves / Eighteen traditionally leafy vegetables consumed as food or medicinal compounds by a majority of people in the KwaZulu Natal province of South Africa were analysed for the presence of potentially harmful chemicals (antinutrients) and for their toxicity and mutagenicity. The purpose of the study was to determine whether leafy vegetables were safe for human consumption. Chemical analysis showed that none of the vegetables contained cyanogenic glycosides, however all the vegetables contained oxalic acid ranging from 24.1 mg/ml to 798.2 mg/ml with Solanum nigrum, Portulaca oleracea and Mormodica balsamina showing the highest concentrations. Most of the vegetables contained negligible amounts of phytic acid and saponins, except for Momordica balsamina (3.01 mg/ml and 1.83 mg/ml, respectively). Fourteen of the plants contained alkaloids with Portulaca oleracea having the highest content (1.53 g total alkaloids/5 g leaf material). Eight of the plants were found to inhibit trypsin activity. These chemical analyses were carried out in duplicate and the mean and standard deviation were used. The Ames test revealed that none of the leafy vegetables produced a mutagenic frequency above 1, except 10 000 µg/ml organic extract of Senna occidentalis (mutagenecity considered at mutagenic frequency above 2), thus none were considered mutagenic. All 18 organic extracts did not kill off more than 50% brine shrimp and were thus considered non-toxic. On the other hand the aqueous extracts of seven vegetables, namely, Physalis viscosa, Amaranthus dubius, Justicia flava, Bidens pilosa, Senna occidentalis, Chenopodium album and Ceratotheca triloba, killed more than 50% of the shrimp and are thus considered toxic above 100 µg/ml. The MTT assay carried out on the organic extracts indicated that 17 vegetables did not kill off more than 50% of HepG2 cells and were thus considered non-cytotoxic. The aqueous extracts of four vegetables, namely, Justicia flava, Asystasia gangetica, Momordica balsamin and Senna occidentalis, however killed more than 50% of the shrimp and were thus considered cytotoxic above 1 000 µg/ml. It may be concluded from the antinutrient analyses and the bioassays on the 18 vegetables that caution needs to be maintained with the consumption of certain leafy vegetables included in this study, especially Senna occidentalis.

Halenia elliptica D. Don belongs to Gentianaceae family. It is often used as part of a traditional Tibetan medicine to treat hepatitis. In the present investigation, six major xanthone components were isolated and identified from Halenia elliptica. An HPLC/DAD/APCI/MS method was developed and validated for the quantitative analysis of these xanthones, including 1-hydroxy-2,3,5-trimethoxy-xanthone (HM-1), 1-hydroxy-2,3,4,7-tetramethoxy- xanthone (HM-2), 1-hydroxy-2,3,4,5-tetramethoxy-xanthone (HM-3), 1,7- dihydroxy-2,3,4,5-tetramethoxy-xanthone (HM-4), 1,5-dihydroxy-2,3-dimethoxy-xanthone (HM-5) and 1-0-[beta-D-xylopyranosyl-(1-6)-beta-D-glucopyranosyl]-2,3,5-trimethoxy-xanthone (HM-2-10). All the xanthones aglycons caused vasodilation in the coronary artery pre-contracted with 1 muM 5-HT, but the xanthone glycoside had no effect. HM-1 was one of the most abundant xanthones with the most potent vasorelaxant activity. / Mechanisms of the vasorelaxant effect of HM-1 were investigated. HM-1 showed a potent vasorelaxant activity on rat coronary artery involved both an endothelium-dependent mechanism involving NO and an endothelium-independent mechanism by inhibiting Ca2+ influx through L-type voltage-operated Ca2+ channels. / Taken together, in spite of the pharmacokinetics results showed that HM-1 was rapidly and widely distributed to tissues after intravenous administration in rats, with conjugation to being the major metabolic pathway in vivo, both HM-1 and its active metabolite (HM-5) show that they are important pharmacological agents with potentially useful therapeutic indications. / The metabolism and pharmacokinetics of HM-1 displayed biphasic elimination kinetics, with an elimination half-life of 60.4 +/- 4.2 min. Four other Phase I metabolites were isolated and identified as demethylated products in vitro. HM-1 was metabolised to HM-5 in the liver. Biliary excretion studies showed that both HM-1 and the metabolite (HM-5) underwent extensive phase II conjugation to form glucuronides and sulfates. Tissue distribution studies showed that HM-1 was widely distributed to different organs. Collection of urine and faeces over 24 h showed that 10.88% of dose was excreted from urine and 1.91% of dose via faeces. / With HM-5 being one of the major in vivo metabolites of HM-1, the effect of HM-5 has been studied on rat coronary artery and compared to HM-1. HM-5-mediated vasorelaxant effect was mediated through opening of potassium channel (TEA, 4-AP) and altering intracellular calcium by partial inhibition of Ca2+ influx through L-type voltage-operated Ca 2+ channels and intracellular Ca2+ stores. / "September 2007." / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4699. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 195-218). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Shum Ka Chiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 174-182). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Table of content --- p.viii / List of figures --- p.xvi / List of tables --- p.xxii / Abbreviations --- p.xxv / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Importance of authentication of Traditional Chinese Medicines --- p.1 / Chapter 1.1.1 --- Confusing nomenclatures --- p.1 / Chapter 1.1.2 --- Similar morphologies of different medicinal materials --- p.2 / Chapter 1.1.3 --- Toxicities of medicinal materials --- p.2 / Chapter 1.1.4 --- Conservation of natural products --- p.2 / Chapter 1.2 --- TCM listed in the Pharmacopoeia of People's Republic of China --- p.3 / Chapter 1.3 --- Overview of mis-use and intoxication of TCM --- p.4 / Chapter 1.4 --- Ordinances regulating Chinese medicines as natural products --- p.7 / Chapter 1.4.1 --- Laws governing Chinese medicine --- p.7 / Chapter 1.4.2 --- Laws governing endangered species --- p.8 / Chapter 1.5 --- Current technologies in the authentication of Traditional Chinese Medicines and their limitations --- p.9 / Chapter 1.6 --- Historical applications of Radix Aconiti --- p.12 / Chapter 1.7 --- Modern applications of Radix Aconiti --- p.16 / Chapter 1.8 --- Research on Radix Aconiti and its chemical components --- p.17 / Chapter 1.8.1 --- Chemistry --- p.17 / Chapter 1.8.2 --- Pharmacology --- p.19 / Chapter 1.8.3 --- Molecular interaction --- p.22 / Chapter 1.9 --- Brief review on the systematics and phylogeny of Aconitum --- p.23 / Chapter 1.10 --- Historical applications of Radix Aucklandiae and related materials --- p.25 / Chapter 1.11 --- Modern applications of Radix Aucklandiae and related material --- p.27 / Chapter 1.12 --- Research on Aucklandiae and related material and their chemical components --- p.28 / Chapter 1.12.1 --- Chemistry --- p.28 / Chapter 1.12.2 --- Pharmacology --- p.29 / Chapter 1.13 --- Brief review on the systematics and phylogeny of Aucklandia and related medicinal species --- p.31 / Chapter 1.14 --- Authentication by DNA sequencing --- p.33 / Chapter 1.14.1 --- Introduction --- p.33 / Chapter 1.14.2 --- Criteria of sequence markers --- p.36 / Chapter 1.14.3 --- Model used to process polymorphism in DNA sequences --- p.37 / Chapter 1.15 --- Screening for novel markers --- p.38 / Chapter 1.15.1 --- Reason for screening novel markers --- p.38 / Chapter 1.15.2 --- Basic principle --- p.39 / Chapter 1.16 --- Introduction to gas chromatography- mass spectrometry --- p.40 / Chapter 1.16.1 --- Basic principles and components of GC-MS --- p.41 / Chapter 1.16.2 --- Advantages and limitations of GC-MS --- p.42 / Chapter 1.16.3 --- Usage of GC-MS on natural product analysis --- p.43 / Chapter 1.16.4 --- Chemometric analysis --- p.44 / Chapter 1.17 --- Objectives --- p.46 / Chapter Chapter 2. --- Materials and Methods --- p.47 / Chapter 2.1 --- Plant samples --- p.47 / Chapter 2.1.1 --- Samples of Aconitum --- p.47 / Chapter 2.1.2 --- Samples of Aucklandia and related species --- p.51 / Chapter 2.2 --- DNA extraction method --- p.58 / Chapter 2.2.1 --- Reagents --- p.58 / Chapter 2.2.2 --- Methods --- p.59 / Chapter 2.3 --- Chemical extraction methods --- p.61 / Chapter 2.4 --- Chemical standard extraction and purification method --- p.62 / Chapter 2.5 --- DNA sequencing --- p.63 / Chapter 2.5.1 --- Reagents --- p.63 / Chapter 2.5.2 --- Methods --- p.65 / Chapter 2.6 --- Genomic subtraction --- p.70 / Chapter 2.7 --- Search for species-specific markers from the subtraction library --- p.74 / Chapter 2.8 --- Gas chromatography- mass spectrometry --- p.74 / Chapter 2.9 --- GC-MS chemometric analysis --- p.75 / Chapter Chapter 3. --- Authentication of Aconitum by DNA Sequencing --- p.76 / Chapter 3.1 --- Introduction --- p.76 / Chapter 3.2 --- Methods --- p.77 / Chapter 3.3 --- Results - 5S spacer --- p.77 / Chapter 3.3.1 --- Sequence information --- p.77 / Chapter 3.3.2 --- Sequence similarity --- p.78 / Chapter 3.3.3 --- Phylogram study --- p.81 / Chapter 3.4 --- Results -psbA-trnH --- p.85 / Chapter 3.4.1 --- Sequence information --- p.85 / Chapter 3.4.2 --- Sequence similarity --- p.85 / Chapter 3.4.3 --- Phylogram study --- p.87 / Chapter 3.5 --- Discussion --- p.91 / Chapter 3.5.1 --- Overview of nuclear ribosomal 5S spacer --- p.91 / Chapter 3.5.2 --- Extensive polymorphism of 5S spacer --- p.91 / Chapter 3.5.3 --- Distribution of samples in the phylograms constructed by 5S spacer --- p.93 / Chapter 3.5.4 --- Utility of 5S spacer for authentication --- p.94 / Chapter 3.5.5 --- Overview of psbA-trnH spacer --- p.94 / Chapter 3.5.6 --- Distribution of samples in the phylograms constructed by psbA-trnH spacer --- p.95 / Chapter 3.5.7 --- A distinctive region of inversion --- p.96 / Chapter 3.5.8 --- Utility of psbA-trnH for authentication --- p.97 / Chapter Chapter 4. --- Screening for Novel Markers for Authentication of Aconitum --- p.98 / Chapter 4.1 --- Introduction --- p.98 / Chapter 4.2 --- Methods --- p.99 / Chapter 4.3 --- Results - subtracted clones --- p.99 / Chapter 4.4 --- Results - SSH6 --- p.104 / Chapter 4.4.1 --- Sequence information --- p.104 / Chapter 4.4.2 --- Sequence similarity --- p.105 / Chapter 4.5 --- Results-SSH15 --- p.107 / Chapter 4.5.1 --- Sequence information --- p.107 / Chapter 4.5.2 --- Sequence similarity --- p.107 / Chapter 4.5.3 --- Phylogram study --- p.109 / Chapter 4.6 --- Results-SSH45 --- p.113 / Chapter 4.6.1 --- Sequence information --- p.113 / Chapter 4.6.2 --- Sequence similarity --- p.113 / Chapter 4.6.3 --- Phylogram study --- p.115 / Chapter 4.7 --- Discussion --- p.119 / Chapter 4.7.1 --- Utility of subtraction in screening markers --- p.119 / Chapter 4.7.2 --- SSH6 --- p.121 / Chapter 4.7.3 --- SSH15 --- p.122 / Chapter 4.7.4 --- SSH45 --- p.123 / Chapter 4.7.5 --- Hybridization in Aconitum --- p.124 / Chapter 4.7.6 --- Inferring species identities of samples from the market --- p.126 / Chapter 4.8 --- Conclusion --- p.128 / Chapter Chapter 5. --- Assessment of Aucklandia lappa and Related Species by GC-MS --- p.129 / Chapter 5.1 --- Introduction --- p.129 / Chapter 5.2 --- Methods --- p.130 / Chapter 5.3 --- Results --- p.130 / Chapter 5.3.1 --- Extraction of essential oil --- p.130 / Chapter 5.3.2 --- GC-MS analysis --- p.131 / Chapter 5.3.3 --- Peak alignment and hierarchical cluster analysis --- p.133 / Chapter 5.3.4 --- Purification of chemical markers from Aucklandia lappa --- p.148 / Chapter 5.3.5 --- Standardization of the purified chemical markers --- p.148 / Chapter 5.3.6 --- Quantitative analysis of chemical markers --- p.152 / Chapter 5.4 --- Discussion --- p.154 / Chapter 5.4.1 --- Analysis of chemical composition --- p.154 / Chapter 5.4.2 --- A comparison on chemometric methods --- p.154 / Chapter 5.4.3 --- Similarity of chemical profiles --- p.156 / Chapter 5.4.4 --- Dendrogram analysis --- p.157 / Chapter 5.4.5 --- Utility of GC-MS in authentication of A. lappa and related species --- p.159 / Chapter 5.4.6 --- Limitations --- p.159 / Chapter 5.4.7 --- Comparison with molecular data --- p.161 / Chapter 5.4.8 --- Contents of dehydrocostuslactone and costunolide --- p.163 / Chapter 5.4.9 --- Locality study --- p.164 / Chapter 5.5 --- Conclusion --- p.165 / Chapter Chapter 6. --- General Discussion --- p.167 / Chapter 6.1 --- DNA sequencing --- p.168 / Chapter 6.2 --- Genomic subtraction --- p.169 / Chapter 6.3 --- Future work on molecular authentication --- p.170 / Chapter 6.4 --- Future work on authentication of Aconitum --- p.170 / Chapter 6.5 --- Gas chromatography- mass spectrometry --- p.171 / Chapter 6.6 --- Future work on authentication by GC-MS --- p.172 / Chapter 6.7 --- Future work on authentication of Aucklandia lappa and related species … --- p.173 / References --- p.174 / Appendix A. Sequence Alignment of 5S Spacer from Aconitum Species --- p.183 / Appendix B. Sequence Alignment of psbA- trnH Spacer from Aconitum Species --- p.188 / Appendix C. Sequences of Subtracted Clones from Aconitum --- p.191 / Appendix D. Sequence Alignment of SSH6 from Aconitum Species --- p.194 / Appendix E. Sequence Alignment of SSH15 from Aconitum Species --- p.195 / Appendix F. Sequence Alignment of SSH45 from Aconitum Species --- p.200 / Appendix G. Gas Chromatograms of Essential Oil Extracts of Aucklandia lappa and Related Species --- p.202

University of Macau / Institute of Chinese Medical Sciences

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 114 leaves / Eighteen traditionally leafy vegetables consumed as food or medicinal compounds by a majority of people in the KwaZulu Natal province of South Africa were analysed for the presence of potentially harmful chemicals (antinutrients) and for their toxicity and mutagenicity. The purpose of the study was to determine whether leafy vegetables were safe for human consumption. Chemical analysis showed that none of the vegetables contained cyanogenic glycosides, however all the vegetables contained oxalic acid ranging from 24.1 mg/ml to 798.2 mg/ml with Solanum nigrum, Portulaca oleracea and Mormodica balsamina showing the highest concentrations. Most of the vegetables contained negligible amounts of phytic acid and saponins, except for Momordica balsamina (3.01 mg/ml and 1.83 mg/ml, respectively). Fourteen of the plants contained alkaloids with Portulaca oleracea having the highest content (1.53 g total alkaloids/5 g leaf material). Eight of the plants were found to inhibit trypsin activity. These chemical analyses were carried out in duplicate and the mean and standard deviation were used. The Ames test revealed that none of the leafy vegetables produced a mutagenic frequency above 1, except 10 000 µg/ml organic extract of Senna occidentalis (mutagenecity considered at mutagenic frequency above 2), thus none were considered mutagenic. All 18 organic extracts did not kill off more than 50% brine shrimp and were thus considered non-toxic. On the other hand the aqueous extracts of seven vegetables, namely, Physalis viscosa, Amaranthus dubius, Justicia flava, Bidens pilosa, Senna occidentalis, Chenopodium album and Ceratotheca triloba, killed more than 50% of the shrimp and are thus considered toxic above 100 µg/ml. The MTT assay carried out on the organic extracts indicated that 17 vegetables did not kill off more than 50% of HepG2 cells and were thus considered non-cytotoxic. The aqueous extracts of four vegetables, namely, Justicia flava, Asystasia gangetica, Momordica balsamin and Senna occidentalis, however killed more than 50% of the shrimp and were thus considered cytotoxic above 1 000 µg/ml. It may be concluded from the antinutrient analyses and the bioassays on the 18 vegetables that caution needs to be maintained with the consumption of certain leafy vegetables included in this study, especially Senna occidentalis.

This study elicits and documents knowledge of the natural environment amongst two rural Black communities in Natal namely, the districts of Maphumulo and Ingwavuma.Twenty members of these communities who are older than 60 years of age were interviewed, as older people are considered by the researcher to be important repositories of environmental knowledge. This study records a variety of animals hunted in these communities and discusses various activities associated with this activity. It examines the gathering and the use of wild edible plants like fruits and spinach, and of wild plants alleged to have medicinal value. It reviews indigenous knowledge related to custom beliefs and prohibitions as well as traditional laws associated .with animals and trees. It also considers how this knowledge can contribute towards the development of Environmental Education in South Africa. The data was deduced from the responses elicited from semi-structured interviews. The data was analyzed qualitatively.