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Identification and characterisation of compounds with antimycobacterial activity from the leaves of Dombeya rotundifolia Hochs

Thesis (M.Sc. (Microbiology)) -- University of Limpopo, 2023 / Plants are not only an important source of medicines, but also play a significant role
in drug development for the treatment of diseases such as Tuberculosis (TB). TB is a
pulmonary disease that is caused by Mycobacterium tuberculosis complex. The aim
of the study is to identify and characterise antimycobacterial compounds from
Dombeya rotundifolia. The plant was collected from the University of Limpopo, dried
and ground into fine powder. Extraction was done using different solvents that differ
in polarity. The plant was screened and analysed for phytochemicals. Three major
phytochemicals were quantified using reagent assays and analysed using standard
curves. The antioxidant activity of the plant was determined using 2,2-Diphenyl-1-
picrylhydrazyl (DPPH) and ferric reducing power assay. The antibacterial activity of
the plant extracts was tested against Mycobacterium smegmatis using bioautography
and serial microplate broth dilution assay. The antibiofilm activity of the plant extracts
were evaluated using crystal violet assay. The anti-inflammatory activity of the plant
was determined using egg albumin protein denaturation assay. The cytotoxic effects
of the extracts were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromide (MTT) assay on THP-1 cell line. The antimycobacterial
compounds were isolated and purified using bioassay-guided assay incorporated with
column chromatography and preparative TLC. The isolated compounds were
analysed and identified using nuclear magnetic resonance spectroscopy (NMR). The
results obtained in this study showed that water was the best extractant, extracting
57.6 mg of the plant material, followed by methanol with 38.1 mg and hexane as the
least extractant with 12.4 mg. In the phytochemical analysis of TLC plates, there was
a separation of compounds in all the mobile systems, and a clear separation of
compounds was observed in the BEA mobile system, followed by the EMW system.
The plant has all major phytochemicals that are needed for drug development. The
plant extracts had antioxidant activity, which was confirmed by the quantitative assays.
The high inhibitory activity of the extracts was indicated by low MIC values that
inhibited the bacterial growth. The butanol extract had the lowest MIC value (0.13
mg/mL). The plant extracts were able to prevent the formation of biofilm at different
concentrations. The plant was proven to have anti-inflammatory properties by having
a high inhibition capacity to prevent protein denaturation. The cytotoxicity results
showed that the plant was toxic to the cells. The isolated antimycobacterial
compounds were identified as Eicosanoic acid and Docosanoic. They have moderate
activity with the MIC value of 0.25 mg/mL. The study indicated that the isolated
compounds have antimycobacterial activity, which showed that the plant has a
potential to treat TB or symptoms related to TB. However, further studies are needed
to evaluate their toxic effects before use. / The National Research Foundation (NRF)

Identiferoai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:ul/oai:ulspace.ul.ac.za:10386/4304
Date January 2023
CreatorsMashilo, Matsilane Lethabo
ContributorsMasoko, P.
Source SetsSouth African National ETD Portal
LanguageEnglish
Detected LanguageEnglish
TypeThesis
Formatxx, 150 leaves
RelationPDF

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