Chemical analysis and biological activities of crude extracts and essential oil of selected medicinal plants from the Eastern Cape, South Africa, and Volta Region of Ghana
- Authors: Agbo, Irene Adzo
- Date: 2023-12
- Subjects: Medicinal plants , Lantana camara , Peptic ulcer -- Treatment , Traditional medicine - South Africa -- Eastern Cape , Traditional medicine -- Ghana
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/62431 , vital:72728
- Description: Lantana camara and Khaya grandifoliola extracts are among many plants found traditionally effective for the treatment of wounds and ulcers. This study assessed the phytochemical content, isolation and identification of single compounds from methanol and ethyl acetate extracts of Lantana camara and Khaya grandifoliola. Further, the bioactivity testing including antioxidant, antimicrobial and cytoxicity of the extracts was done to confirm the wound healing potential discovered by the traditional healers. Materials and methods: Extraction was done successively using maceration method with 100 % ethyl acetate and 100 % methanol with a biomass-to-solvent ratio of 1:3 (w/v) to obtain L. camara ethyl acetate extracts of berry (ELB), flower (ELF) and leaf (ELL) and methanol extracts of MLB, MLF, MLL and K. grandifoliola ethyl acetate extracts of leaf (EKL), root (EKR) and stem bark (EKSB) and methanol extracts of MKL, MKR, MKSB respectively. L. camara leaf essential oil (EO) was extracted using the hydro-distillation method with a Clevenger apparatus. Total phytochemical content was assessed for each extract using spectrophotometric methods and a calibration curve of standards: bromocresol green method with atropine; Folin–Ciocalteu colorimetric method with gallic acid, aluminium chloride colorimetric method with quercetin and concentrated sulphuric acid chloroform with linalool for total alkaloid, phenolic, flavonoid and terpenoid contents respectively. Single compound isolation and purification was conducted using chromatographic techniques. Elucidation of single compounds was done using spectrometric method, high resolution- mass spectrometry, and one and two-dimensional (1D and 2D)-NMR. Stereochemistry of each compound was confirmed using electronic circular dichroism spectra. A Crystalline compound was identified by single crystal X-ray diffraction using CuKα-radiation. In vitro bioactivities were assessed with methods such as 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide, free radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl, inhibitory effect on nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages, and 96-well plate micro dilution for cytotoxicity, ant-inflammatory and antimicrobial activity testing. Results: Methanol extracts of both plants retained high phytochemical concentrations of all the phytoconstituents investigated compared with the ethyl acetate extracts which retained lower concentrations. The results of the L. camara methanol extracts include; total alkaloid content (TAC) (2.05±0.18, 1.87±1.54 and 2.60±1.10 mg AEQ/100 mg); total phenolic content (TPC) (14.05±4.04, 34.59±3.01 and 18.58±1.87 mg GAEQ/100 mg); the total flavonoid content(TFC) of flower (12.45±1.87, 20.41±2.69 and mg QEQ/100 mg); total terpenoids (TTC) (20.74±2.34, 20.74±2.34 and 15.97±1.19 mg LIN EQ/100mg) of MLB, MLF and MLL respectively. Whereas that of the K. grandifoliola methanol extracts include; TAC (7.32±0.14,8.49±0.34, 10.67±0.22 mg AEQ/100 mg); TPC (37.49±1.40, 44.41±0.69, 53.57±1.50 mgGAEQ/100 mg); TFC (6.54±0.55, 9.58±0.89 and 10.26±0.92 mg QEQ/100 mg); TTC(10.16±1.41, 35.78±2.14 and 23.45±1.76 mg LIN EQ/100mg) of MKL, MKR and MKSB respectively. The major components of essential oil, out of the 71 constituents identified include Davanone D (32.91 %), Caryophyllene (5.07 %), Nerolidol 2 (3.56 %) and GermacreneD (3.13 %). Compounds 3.47 was isolated from the methanol extract of L. camara flowers. This compound is reported for the first time from the L. camara flower extract. Two compounds, compounds 4.23, and 4.26, were isolated from the methanol extract of K. grandifoliola roots, compound 4.22 was isolated from the ethyl acetate root extract while compounds 4.24 and 4.25 were isolated from the ethyl acetate stem bark extract as isomers in a mixture. Compounds 4.22 and 4.23 are reported from K. grandifoliola root for the first time. The isolated compounds (compounds 3.47 and 4.23) were nontoxic to the Vero cell line and this may contribute to possible stimulation of cell proliferation, promoting wound healing. Cytotoxicity describes extract virulence to Vero cell line. MLF and ELB were found nontoxic even at the highest concentration of 200 μg/mL. The MKSB and MKR, as well as the EKSB were nontoxic. Antioxidant activity results, described by the percentage inhibition in the DPPH assay, showed that MLF and MKSB had the highest antioxidant activities compared with the ascorbic acid standard, with IC50 of 38.68±5.09 and 37.03±11.95 μg/mL for L. camara and K. grandifoliola respectively. ELB exhibited a significant anti-inflammatory activity inhibiting NO• radical generation in the LPS-stimulated RAW 264.7 macrophages at concentration ranging from 50 and 100 μg/mL. EKSB and MKR showed significant anti-inflammatory activity at 100 and 200 μg/ml respectively. ELL and ELF demonstrated potent growth inhibition against S. pyogenes with an MIC value ≤ 0.125 mg/mL, while the MICs of the ELB and MLL were 0.5 mg/mL and 2 mg/mL respectively. MKSB and MKR and EKSB extract exhibited an effective growth inhibition against S. aureus with MIC of 1 mg/mL. The growth of S. pyogenes was supressed by both ethyl acetate and methanol extracts of all plant parts tested with MIC ranging from 0.25–2 mg/mL. Conclusion: The potent bioactivity shown in the results of the cytotoxicity, antioxidant activity, anti-inflammatory and antimicrobial activity testing, and the nontoxic singlecompounds of L. camara and K. grandifoliola extracts led to the conclusion that the two plants had wound healing potential. The study therefore confirmed their traditional uses for treatment of wounds. , Thesis (PhD) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-12
- Authors: Agbo, Irene Adzo
- Date: 2023-12
- Subjects: Medicinal plants , Lantana camara , Peptic ulcer -- Treatment , Traditional medicine - South Africa -- Eastern Cape , Traditional medicine -- Ghana
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/62431 , vital:72728
- Description: Lantana camara and Khaya grandifoliola extracts are among many plants found traditionally effective for the treatment of wounds and ulcers. This study assessed the phytochemical content, isolation and identification of single compounds from methanol and ethyl acetate extracts of Lantana camara and Khaya grandifoliola. Further, the bioactivity testing including antioxidant, antimicrobial and cytoxicity of the extracts was done to confirm the wound healing potential discovered by the traditional healers. Materials and methods: Extraction was done successively using maceration method with 100 % ethyl acetate and 100 % methanol with a biomass-to-solvent ratio of 1:3 (w/v) to obtain L. camara ethyl acetate extracts of berry (ELB), flower (ELF) and leaf (ELL) and methanol extracts of MLB, MLF, MLL and K. grandifoliola ethyl acetate extracts of leaf (EKL), root (EKR) and stem bark (EKSB) and methanol extracts of MKL, MKR, MKSB respectively. L. camara leaf essential oil (EO) was extracted using the hydro-distillation method with a Clevenger apparatus. Total phytochemical content was assessed for each extract using spectrophotometric methods and a calibration curve of standards: bromocresol green method with atropine; Folin–Ciocalteu colorimetric method with gallic acid, aluminium chloride colorimetric method with quercetin and concentrated sulphuric acid chloroform with linalool for total alkaloid, phenolic, flavonoid and terpenoid contents respectively. Single compound isolation and purification was conducted using chromatographic techniques. Elucidation of single compounds was done using spectrometric method, high resolution- mass spectrometry, and one and two-dimensional (1D and 2D)-NMR. Stereochemistry of each compound was confirmed using electronic circular dichroism spectra. A Crystalline compound was identified by single crystal X-ray diffraction using CuKα-radiation. In vitro bioactivities were assessed with methods such as 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide, free radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl, inhibitory effect on nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages, and 96-well plate micro dilution for cytotoxicity, ant-inflammatory and antimicrobial activity testing. Results: Methanol extracts of both plants retained high phytochemical concentrations of all the phytoconstituents investigated compared with the ethyl acetate extracts which retained lower concentrations. The results of the L. camara methanol extracts include; total alkaloid content (TAC) (2.05±0.18, 1.87±1.54 and 2.60±1.10 mg AEQ/100 mg); total phenolic content (TPC) (14.05±4.04, 34.59±3.01 and 18.58±1.87 mg GAEQ/100 mg); the total flavonoid content(TFC) of flower (12.45±1.87, 20.41±2.69 and mg QEQ/100 mg); total terpenoids (TTC) (20.74±2.34, 20.74±2.34 and 15.97±1.19 mg LIN EQ/100mg) of MLB, MLF and MLL respectively. Whereas that of the K. grandifoliola methanol extracts include; TAC (7.32±0.14,8.49±0.34, 10.67±0.22 mg AEQ/100 mg); TPC (37.49±1.40, 44.41±0.69, 53.57±1.50 mgGAEQ/100 mg); TFC (6.54±0.55, 9.58±0.89 and 10.26±0.92 mg QEQ/100 mg); TTC(10.16±1.41, 35.78±2.14 and 23.45±1.76 mg LIN EQ/100mg) of MKL, MKR and MKSB respectively. The major components of essential oil, out of the 71 constituents identified include Davanone D (32.91 %), Caryophyllene (5.07 %), Nerolidol 2 (3.56 %) and GermacreneD (3.13 %). Compounds 3.47 was isolated from the methanol extract of L. camara flowers. This compound is reported for the first time from the L. camara flower extract. Two compounds, compounds 4.23, and 4.26, were isolated from the methanol extract of K. grandifoliola roots, compound 4.22 was isolated from the ethyl acetate root extract while compounds 4.24 and 4.25 were isolated from the ethyl acetate stem bark extract as isomers in a mixture. Compounds 4.22 and 4.23 are reported from K. grandifoliola root for the first time. The isolated compounds (compounds 3.47 and 4.23) were nontoxic to the Vero cell line and this may contribute to possible stimulation of cell proliferation, promoting wound healing. Cytotoxicity describes extract virulence to Vero cell line. MLF and ELB were found nontoxic even at the highest concentration of 200 μg/mL. The MKSB and MKR, as well as the EKSB were nontoxic. Antioxidant activity results, described by the percentage inhibition in the DPPH assay, showed that MLF and MKSB had the highest antioxidant activities compared with the ascorbic acid standard, with IC50 of 38.68±5.09 and 37.03±11.95 μg/mL for L. camara and K. grandifoliola respectively. ELB exhibited a significant anti-inflammatory activity inhibiting NO• radical generation in the LPS-stimulated RAW 264.7 macrophages at concentration ranging from 50 and 100 μg/mL. EKSB and MKR showed significant anti-inflammatory activity at 100 and 200 μg/ml respectively. ELL and ELF demonstrated potent growth inhibition against S. pyogenes with an MIC value ≤ 0.125 mg/mL, while the MICs of the ELB and MLL were 0.5 mg/mL and 2 mg/mL respectively. MKSB and MKR and EKSB extract exhibited an effective growth inhibition against S. aureus with MIC of 1 mg/mL. The growth of S. pyogenes was supressed by both ethyl acetate and methanol extracts of all plant parts tested with MIC ranging from 0.25–2 mg/mL. Conclusion: The potent bioactivity shown in the results of the cytotoxicity, antioxidant activity, anti-inflammatory and antimicrobial activity testing, and the nontoxic singlecompounds of L. camara and K. grandifoliola extracts led to the conclusion that the two plants had wound healing potential. The study therefore confirmed their traditional uses for treatment of wounds. , Thesis (PhD) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-12
The suitability of Alagoasa extrema Jacoby (Coleoptera: Chrysomelidae: Alticinae), as a biological control agent for Lantana camara L. in South Africa
- Authors: Williams, Hester Elizabeth
- Date: 2003
- Subjects: Lantana camara , Lantana camara -- South Africa , Biological pest control agents -- South Africa , Chrysomelidae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5783 , http://hdl.handle.net/10962/d1005471 , Lantana camara , Lantana camara -- South Africa , Biological pest control agents -- South Africa , Chrysomelidae
- Description: Lantana camara Linnaeus (Verbenaceae), commonly known as lantana, is a highly invasive weed in many parts of the world. In South Africa it is naturalized in several provinces where it invades pastures, riverbanks, mountain slopes and valleys and commercial and natural forests, forming dense, impenetrable thickets. Chemical and mechanical control methods are expensive, labour intensive and provide only temporary relief as cleared areas are rapidly reinfested by seedlings and coppice growth. A biological control programme was initiated in South Africa in the 1960s, but despite the establishment of 11 agent species, it was considered to have had limited success. Several factors are thought to restrict the impact of the biocontrol agents. Firstly, L. camara occurs in a range of climatic regions, some of which are unsuitable for the establishment of agent species of tropical and subtropical origin. Secondly, L. camara is the result of hybridization between several Lantana species, forming a complex of hybridized and hybridizing varieties in the field, which match none of the Lantana species in the region of origin. This causes partial insect-host incompatibility, displayed as varietal preference. Thirdly, parasitism appears to have significantly reduced the effectiveness of several natural enemies. In spite of all these constraints, biological control has reduced invasion by L. camara by 26%. However, the weed is still very damaging and additional natural enemies are required to reduce infestations further. A flea-beetle species, Alagoasa extrema Jacoby (Coleoptera: Chrysomelidae), was collected from several sites in the humid subtropical and tropical regions of Mexico, and imported into quarantine in South Africa and studied as a potential biocontrol agent for L. camara. Favourable biological characteristics of this beetle included long-lived adults, several overlapping generations per year, and high adult and larval feeding rates. Observations from the insect’s native range and studies in South Africa suggest that A. extrema would probably be more suited to the subtropical, rather than the temperate areas in South Africa. Laboratory impact studies indicated that feeding damage by A. extrema larvae, over a period spanning the larval stage (16 to 20 days), reduced the above-ground biomass of L. camara plants by up to 29%. Higher larval populations resulted in a higher reduction of biomass. Varietal preference and suitability studies indicated that A. extrema exhibits a degree of varietal preference under laboratory conditions, with one of the white pink L. camara varieties proving the most suitable host. This variety is one of the most damaging varieties in South Africa and is particularly widespread in Mpumalanga Province. Although A. extrema proved to be damaging to L. camara, laboratory host range trials showed it to be an oligophagous species, capable of feeding and developing on several non-target species, especially two native Lippia species (Verbenaceae). The host suitability of these species was marginally lower than that of L. camara and the potential risk to these indigenous species was deemed to be too high to warrant release. It was therefore recommended that A. extrema not be considered for release in South Africa.
- Full Text:
- Date Issued: 2003
- Authors: Williams, Hester Elizabeth
- Date: 2003
- Subjects: Lantana camara , Lantana camara -- South Africa , Biological pest control agents -- South Africa , Chrysomelidae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5783 , http://hdl.handle.net/10962/d1005471 , Lantana camara , Lantana camara -- South Africa , Biological pest control agents -- South Africa , Chrysomelidae
- Description: Lantana camara Linnaeus (Verbenaceae), commonly known as lantana, is a highly invasive weed in many parts of the world. In South Africa it is naturalized in several provinces where it invades pastures, riverbanks, mountain slopes and valleys and commercial and natural forests, forming dense, impenetrable thickets. Chemical and mechanical control methods are expensive, labour intensive and provide only temporary relief as cleared areas are rapidly reinfested by seedlings and coppice growth. A biological control programme was initiated in South Africa in the 1960s, but despite the establishment of 11 agent species, it was considered to have had limited success. Several factors are thought to restrict the impact of the biocontrol agents. Firstly, L. camara occurs in a range of climatic regions, some of which are unsuitable for the establishment of agent species of tropical and subtropical origin. Secondly, L. camara is the result of hybridization between several Lantana species, forming a complex of hybridized and hybridizing varieties in the field, which match none of the Lantana species in the region of origin. This causes partial insect-host incompatibility, displayed as varietal preference. Thirdly, parasitism appears to have significantly reduced the effectiveness of several natural enemies. In spite of all these constraints, biological control has reduced invasion by L. camara by 26%. However, the weed is still very damaging and additional natural enemies are required to reduce infestations further. A flea-beetle species, Alagoasa extrema Jacoby (Coleoptera: Chrysomelidae), was collected from several sites in the humid subtropical and tropical regions of Mexico, and imported into quarantine in South Africa and studied as a potential biocontrol agent for L. camara. Favourable biological characteristics of this beetle included long-lived adults, several overlapping generations per year, and high adult and larval feeding rates. Observations from the insect’s native range and studies in South Africa suggest that A. extrema would probably be more suited to the subtropical, rather than the temperate areas in South Africa. Laboratory impact studies indicated that feeding damage by A. extrema larvae, over a period spanning the larval stage (16 to 20 days), reduced the above-ground biomass of L. camara plants by up to 29%. Higher larval populations resulted in a higher reduction of biomass. Varietal preference and suitability studies indicated that A. extrema exhibits a degree of varietal preference under laboratory conditions, with one of the white pink L. camara varieties proving the most suitable host. This variety is one of the most damaging varieties in South Africa and is particularly widespread in Mpumalanga Province. Although A. extrema proved to be damaging to L. camara, laboratory host range trials showed it to be an oligophagous species, capable of feeding and developing on several non-target species, especially two native Lippia species (Verbenaceae). The host suitability of these species was marginally lower than that of L. camara and the potential risk to these indigenous species was deemed to be too high to warrant release. It was therefore recommended that A. extrema not be considered for release in South Africa.
- Full Text:
- Date Issued: 2003
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