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Pharmacological investigation of some trees used in South African traditional medicine.Eldeen, Ibrahim Mohamed Suliman. January 2005 (has links)
South Africa is home to a wide diversity of cultural groups, all of which
utilize the flora for a variety of purposes. This is true with regard to traditional
medicine systems which are similar to those of the rest of Africa south of the
Sahara, with diviners (sangomas) and herbalists (inyangas) as the key health
providers. In addition, the Country is rich in plant diversity with some 30 000
species of flowering plants - almost one tenth of the worlds recorded higher plants.
This incorporates a large diversity of plants including trees, shrubs, herbs, bulbs
and corms.
The adverse effects of traditional medicinal plants and natural products
are not well documented in the literature. Recently, many plants used as food or in
traditional medicine have been shown to be potentially mutagenic using in vitro
assays. Thus, the scientific evaluation of traditional medicine and medicinal plants
is very important to validate claims made on safety and efficiency of such usages.
After a survey of the available ethnobotanical literature, ten trees used in
South African traditional medicine were selected. These species were: Acacia
niolotica subspecies kraussiana, Acacia sieberiana, Albizia adianthifolia,
Combretum kraussii, Faidherbia albida, Ficus sur, Prunus africana, Salix
mucronata, Terminalia sericea and Trichilia dregeana. Plant parts including leaf,
root and bark were collected from each of the selected trees (exceptions were
Albizia adianthifolia, Faidherbia albida, Terminalia sericea and Prunus africana)
and extracted using ethyl acetate, ethanol and water individually to ensure the
extraction of compounds over a wide range of polarities. The extracts (in total, 78)
were screened for antibacterial, anti-inflammatory (COX-1 and COX-2) and antiacetylcholinesterase
activities and investigated for their potential mutagenic effects
using the Ames test.
Antibacterial activity was detected using the disc-diffusion and microdilution
assays. The extracts were tested against Gram-positive bacteria: Bacillus
subtilis, Staphylococcus aureus, Micrococcus luteus and Gram-negative bacteria:
Escherichia coli and Klebsiella pneumoniae. Of the 78 different plant extracts
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tested (final amount of plant material was 1 mg per disc), 84% showed activity
against Gram-positive bacteria. From this percentage, 20% also showed activity
against Gram-negative bacteria. The best inhibition was observed with ethyl
acetate and ethanol root extracts of Terminalia sericea against both Gram-positive
and Gram-negative bacteria.
In the micro-dilution assay, 55% of the plant extracts showed minimum
inhibitory concentration (MIC) values ~ 1.56 mg/ml against Gram-positive and/or
Gram-negative bacteria. The ethyl acetate bark extract of Acacia sieberiana and
the root and bark ethyl acetate extracts of Acacia nilotica inhibited bacterial growth
of both Gram-positive and Gram-negative bacteria at concentrations ~ 0.8 mg/ml.
The aqueous leaf extracts of Acacia sieberiana had a low MIC value (0.3 mg/ml)
against Gram-negative Kleibsiella pneumoniae and the ethyl acetate extracts of the
root inhibited growth of Escherichia coli with an MIC value of 0.1 mg/ml. However,
these two extracts showed no activity in the disc-diffusion assay. The MIC values of
the neomycin (control) were 0.8 I-Ig/ml and 3.1 I-Ig/ml against Kleibsiella
pneumoniae and Escherichia coli respectively.
In the anti-inflammatory test, 70% of the plant extracts from different plant
parts (leaf, root, bark) of the tree investigated showed strong inhibition in both the
CQX-1 and CQX-2 bioassays. The CQX-2 inhibitory effects of aqueous extracts
were generally lower when compared to the organic solvent extracts. However,
water extracts of Acacia nilotica was an exception (~ 90%).
In the acetylcholinesterase inhibitory test, 21% of the plant extracts were
active at concentrations ~ 1 mg/ml using the micro-plate assay. The lowest IC50
value was 0.04 mg/ml obtained with an ethanol bark extract of Combretum kraussii.
The IC50 value of the galanthamine (positive control) was 2 I-IM.
None of the investigated plants showed any potential mutagenic effects
with Salmonella typhymurium strain TA 98 using the Ames test.
Using bioassay-guided fractionation, anolignan B was isolated from the
ethyl acetate root extract of Terminalia sericea. Antibacterial activity of anolignan B
was determined using the microdilution assay. The compound possessed activity
against both Gram-positive and Gram-negative bacteria. The lowest MIC value
(3.8 IJg/ml) was observed with Staphylococcus aureus. MIC value of the neomycin
was 1.5 IJg/ml.
Anti-inflammatory activity of anolignan B was detected using the CQX-1
and CQX-2 bioasays. The compound showed strong inhibitory activity against
CQX-1 and weaker activity against CQX-2. The ICso values were 1.5 mM and 7.5
mM with CQX-1 and CQX-2 respectively. The ICso values of indomethacin were
0.003 mM and 0.186 mM against CQX-1 and CQX-2 respectively.
There were no potential mutagenic effects showen by anolignan B against
Salmonella typhimurium strain TA 98 in the Ames test.
Isolation of anolignan B from Terminalia species and the antibacterial and
anti-inflammatory activities observed in this work have not been reported
previously and could therefore be recorded as novel biological activities for this
compound. These results also support the idea that the use of ethnobotanical data
can provide a valuable short cut by indicating plants with specific uses which might
likely be sources of biologically active chemicals. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.
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Potencial invasor do chapéu-de-sol (Terminalia catappa L.) em área de restinga / Invading potencial from Singapure almond (Terminalia catappa L.) in resting forestJoyce Helena Sanches 20 February 2009 (has links)
A atual e a próxima década serão identificadas no tempo como fórum das mais relevantes preocupações com a qualidade ambiental do planeta. Temas como aquecimento global e a eminente perda da quantidade e qualidade da água são discutidos constantemente. Um outro problema ambiental mais específico tem preocupado especialistas no mundo todo, incluindo o Brasil. Trata-se da disseminação em grande escala de espécies exóticas. O tema tem sido tratado no meio científico e acadêmico como invasões biológicas e não se restringe somente às plantas.O estudo aqui apresentado trata de uma espécie arbórea com grande disseminação no país e utilizada em áreas urbanas e faixas de praia devido à sombra proporcionada na época do verão. A ocorrência do chapéu-de-sol em unidades de conservação de proteção integral pode representar uma ameaça às espécies autóctones pela ocupação do espaço. A identificação com metodologia científica das características germinativas e sucesso reprodutivo foram a forma de avaliar o grau de ameaça que a espécie representa para o ambiente de restinga.Assim, esse estudo teve o objetivo de determinar o potencial de contaminação biológica do chapéu-de-sol (Terminalia catappa L.) através da análise do índice de velocidade de germinação (IVG), tempo médio de germinação, velocidade média de germinação, freqüência germinativa, germinabilidade (G%) e análise da taxa de sobrevivência e de transição de plântulas, jovens e adultos. Os resultados indicam que as sementes da espécie podem germinar em ampla faixa de temperatura, principalmente, se estiver a pleno sol. Frutos despolpados apresentaram maior velocidade e taxa de germinação.Plântulas apresentaram elevada taxa de mortalidade comparada aos indivíduos jovens e adultos. Concluiu-se que a espécie precisa ser retirada da área de restinga, pois, apesar de ser considerada estabelecida, em condições favoráveis ela se desenvolverá ocupando o nicho de outras espécies locais. / Current and the next decade they will be identified in the time as group of the most excellent concerns with the ambient quality of the planet. Subjects as global heating and the eminent loss of the amount and quality of the water are argued constantly. A more specific ambient problem has worried specialists in the world all, including Brazil. One is about the large-scale dissemination of exotic species. The subject has been treated in the half academic as biological invasions and it is not only restricted to the plants. The study presented here it at the time deals with a trees species with great dissemination in the country and used in urban areas and beach bands due to proportionate shade of the summer. The occurrence of the singapure almond in protect areas of integral protection can represent a threat to the species for the occupation of the space. The identification with scientific methodology of the germination characteristics and reproductive success had been the form to evaluate the threat degree that the species represents for the environment of restinga. Thus, this study it had the objective to determine the potential of biological contamination of the singapure almond (Terminalia catappa L.) through the analysis of the index of germination speed (IVG), average time of germination, average speed of germination, frequency of germination, germinability (G%) and analysis of the tax of survival and transistion of seedly, young and adults. The results indicate that the seeds of the species can germinate in ample band of temperature, mainly, will be the full sun. Pulped fruits had presented greater speed and tax of germination. Seedly had presented high tax of comparative mortality to adult and youngs. The necessary species was concluded that to be removed of the area of restinga, therefore, although to be considered established, in favorable conditions it will develop itself occupying the niche of other local species.
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Phytochemical, biological and toxicity studies of terminalia sericea burch. (Combretaceae)Anokwuru, Chinedu Prosper 18 May 2018 (has links)
PhD (Chemistry) / Department of Chemistry / Terminalia sericea Burch. ex. DC (Combretaceae) is one of the 50 most popular medicinal plants in Africa. The fruit, leaves, stems and roots are commonly used for the treatment of cough, skin infections, diabetes, diarrhoea, venereal diseases and tuberculosis. However, the roots are most commonly used in the preparation of traditional medicines. Pharmacological studies have revealed that the crude root extracts display antibacterial activity against several Gram-positive and Gram-negative bacteria. Anolignan b, termilignan b and arjunic acid are reported to be the major antibacterial constituents present in the roots. Other compounds isolated from the roots include resveratrol-3-rutinoside, sericic acid, sericoside and arjunglucoside I.
Authorities worldwide, including the Medicines Control Council of South Africa, have begun to regulate herbal drugs sold in the form of commercial formulations. Quality control of herbal drugs is challenging, since the chemical profiles of the raw materials may vary, depending on the origin of the plant material and the way that it was handled and processed. The chemistry, in turn, impacts on the safety and efficacy of the plant material. To date, there are no available data on parameters that can be used to standardise the quality of T. sericea raw materials. The aim of this study was therefore to provide information on the variation of the chemical constituents that contribute to the biological effects of the roots of T. sericea and also establish its safety.
Since the compounds previously isolated from the roots were not commercially available, isolation of the major constituents of the roots was undertaken to obtain analytical standards. A crude dichloromethane:methanol (1:1) extract was initially fractionated using silica gel column chromatography, where after, some of the fractions were further purified using silica gel and Sephadex LH-20 column chromatography. Final purification of the enriched fractions was achieved using preparative high performance liquid chromatography coupled with mass spectrometry (prep-HPLC-MS). The structures of these compounds were subsequently elucidated using one- and two- dimensional nuclear magnetic
resonance (NMR) spectroscopy and mass spectrometry and identified as sericic acid (340 g), sericoside (500 g), resveratrol-3-rutinoside (240 mg) and arjunglucoside I (74 mg).
The chemical variation within the crude root extracts of samples (n = 42) from ten populations in the Limpopo Province of South Africa, was determined using ultra performance liquid chromatography-mass spectrometry (UPLC-MS). A method was developed for the simultaneous determination of sericic acid, resveratrol-3-rutinoside, sericoside and arjungluicoside I in the extracts using UPLC with photodiode array detection (PDA). The method was validated according to the guidelines of the International Council for Harmonisation (ICH). A regression coefficient (R2) of 0.998 was obtained for sericic acid, resveratrol-3- rutinoside and arjunglucoside I, while the R2 value for sericoside was 0.999, indicating a linear relationship between the concentration and the detector response. Satisfactory limits of detection for sericic acid (25.2 ng/mL), sericoside (11.6 ng/mL), resveratrol-3-rutinoside (23.3 ng/mL) and arjunglucoside I (8.81 ng/mL) were determined. Recoveries of 98 % and 80% were obtained for samples spiked with 12.5 μg/mL and 25 μg/mL of resveratrol-3-rutinoside, respectively, indicating that the method is accurate. The intra- and inter-day variation in resveratrol-3-rutinoside concentration, measured over three days, indicated excellent analytical precision, since all the relative standard deviations were below 0.70 %. The quantitative data revealed that sericic acid (1.59 to 8.45 mg/g), sericoside (2.07 to 20.17 mg/g), resveratrol-3-rutinoside (0.65 to 29.82 mg/g) and arjunglucoside I (0.86 to 8.44 mg/g dry weight) were the major constituents of the root samples, but their concentrations were highly variable.
Chemometric analysis of the aligned UPLC-MS data was used to investigate similarities and differences in the chemical profiles of the samples using an untargeted approach. A principal component analysis (PCA) model was constructed and subsequently hierarchical cluster analysis (HCA) indicated the presence of two main groups, which were found to be independent of the populations to which the samples belong. Classes, based on the HCA class identifiers, were subsequently assigned to the samples, and an orthogonal projection to latent structures-discriminant analysis (OPLS-DA) model was then constructed, (R2 cum = 0.996 and Q2 cum = 0.967). The corresponding loadings
plot allowed sericic acid, sericoside and resveratrol-3-rutinoside to be identified as biomarkers associated with the first group. Quantitative, rather than qualitative differences were responsible for the observed clustering pattern. Techniques that could be applied in quality control protocols for T. sericea root were investigated.
High performance thin layer chromatography (HPTLC) analysis of the root extracts was optimised by testing different developing solvents and visualization reagents. The presence of the sericic acid (Rf = 0.80), sericoside (Rf = 0.49) and resveratrol-3-rutinoside (Rf = 0.36) were clearly visible on the plates. There were visible variations in the concentrations of resveratrol-3-rutinoside in representative samples from the 10 populations, corresponding to the UPLC results. The powdered samples were then analysed by mid-(MIR) infrared spectroscopy. Chemometric analysis of the data revealed no definitive clustering pattern. Partial least squares-discriminant analysis (PLS-DA) calibration models were established from the MIR spectral data combined with the accurate UPLC-values, for the prediction of the sericoside (R2Y = 0.848, Q2 = 0.757, RMSEP = 2.70 mg/g) and resveratrol-3-rutinoside (R2Y = 0.794, Q2 = 0.695, RMSEP = 4.37 mg/g) concentrations in powdered root samples.
The antibacterial activities of the root extracts, column fractions and isolated compounds were determined using three Gram-positive and five Gram-negative bacteria, all selected due to their ability to cause intestinal and skin disorders. Extracts and fractions containing high concentrations of sericic acid exhibited the highest activities against Klebsiella pneumoniae (ATCC 13883), Pseudomonas aeruginosa (ATCC 27858), Salmonella typhimurium (ATCC 14028), Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 12223) and Shigella sonnei (ATCC 9292). The pure compound (sericic acid) was highly active against S. sonnei (MIC 0.078 μg/mL), a Gram- negative bacterium. There were no variations in the activity of the crude extracts against B. cereus and P. aeruginosa, while the MIC values obtained against S. typhi were variable and ranged from 0.25 to 1.0 mg/mL. Sericoside and resveratrol-3-rutinoside did not display any activity.
The anti-oxidant activities were evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) and reducing power assays. The anti-oxidant assays revealed that resveratrol-3- rutinoside exhibited lower activity (DPPH = 186 μg/mL; RP = 184 μg/mL) compared to the crude extract (DPPH = 22.3 μg/mL; RP = 24.4 μg/mL) and ascorbic acid (DPPH = 11.3 μg/mL, RP = 145 μg/mL). Sericic acid and sericoside did not display any anti- oxidant activities. The variation in the anti-oxidant activities (4.58 to 26.0 μg/mL) of the samples from different populations was an indication of chemical variability.
A toxicity study of the raw powdered plant material was conducted using vervet monkeys (Chlorocebus pygerythrus). Biochemical analysis (liver function tests, kidney function tests and hematology), physical and physiological examinations were conducted. The subjects were fed a normal diet supplemented with T. sericea root powder (2.14 g/kg per day) for 120 days, where after the diet was returned to normal (washout) for another 30 days. The treatment groups presented with elevated serum enzymes at Week 4, followed by the reduction of the elevated serum enzymes levels at Week 12. These results indicate short-term hepatotoxic effects, followed by hepatoprotective activity. Reduction of the serum glucose at Week 4 suggests hypoglycemic potential. However, elevated serum creatinine levels indicated possible nephrotoxicity.
In conclusion, this study has indicated the variability in the chemical constituents of the roots of T. sericea, which affects the antibacterial and anti-oxidant activities. Sericic acid, resveratrol-3-rutinoside, and sericoside were, for the first time, identified as biomarkers that can be used for the quality control of raw root material to be used in herbal products. Sericic acid was also found to be the main antibacterial constituent of the roots. The hepatoprotective, nephrotoxic and hematotoxic effects observed in monkeys to which the root powder had been administered is cause for concern. / NRF
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