Dissertations / Theses on the topic 'Phytochemical analysis'

Thesis (Ph. D.)--City University of New York, 2007.
Includes bibliographical references (leaves 100-120) and index. Also available for educational and research purposes in PDF format on the Internet.

In Africa, the importance of medicinal plants in folklore medicine and their contribution to primary healthcare is well recognized. Across the continent, local herbal mixtures still provide the only therapeutic option for about 80% of the population. The vast floral diversity and the intrinsic ethnobotanical knowledge has been the backbone of localized traditional herbal medical practices. In Africa, an estimated 5400 of the 60000 described plant taxa possess over 16300 therapeutic uses. Similarly, with a therapeutic flora comprising of approximately 650 species, herbal medical practitioners in South Africa, make use of a plethora of plants to treat different human diseases and infections. Over the years, studies have identified numerous plant species with potential against chronic metabolic diseases including type 2 diabetes mellitus (T2DM). Globally, the incidence and prevalence of T2DM have reached epidemic proportions affecting people of all ages, nationalities and ethnicity. Considered the fourth leading cause of deaths by disease, T2DM is a global health crisis with an estimated diagnosis and mortality frequency of 1 every 5 seconds and 1 every 7 seconds respectively. Though the exact pathophysiology of T2DM is not entirely understood, initial peripheral insulin resistance in adipose tissue, liver, and skeletal muscle with subsequent pancreatic β-cell dysfunction resulting from an attempt to compensate for insulin resistance is a common feature of the disease. The current approach to treating T2DM is the use of oral antidiabetic agents (OAAs), insulin, and incretin-based drugs in an attempt to achieve glycaemic control and maintain glucose homeostasis. However, conventional anti-T2DM drugs have been shown to have limited efficacies and serious adverse effects. Hence, the need for newer, more efficacious and safer anti-T2DM agents. Sutherlandia frutescens subsp. microphylla is a flowering shrub of the pea family (Fabaceae/Leguminaceae) found mainly in the Western Cape and Karoo regions of Southern Africa. Concoctions of various parts of the plant are used in the management of different ailments including T2DM. However, despite extensive biological and pharmacological studies, few analyses exist of the chemical constituents of S. frutescens and no Triple Time of Flight Liquid Chromatography with Mass Spectrometry (Triple TOF LC/MS/MS) analysis has been performed. The initial aim of this study was to investigate the phytochemical profile of hot aqueous, cold aqueous, 80% ethanolic, 100% ethanolic, 80% methanolic and 100% methanolic extracts of a single source S. frutescens plant material using colorimetric and spectrophotometric analysis. The hot aqueous extractant was found to be the best extractant for S. frutescens, yielding 1.99 g of crude extract from 16 g fresh powdered plant material. This data suggests that application of heat and water as the extractant (hot aqueous) could play a vital role in extraction of bioactive compounds from S. frutescens and also justifies the traditional use of a tea infusion of S. frutescens. Colorimetric analysis revealed the presence of flavonoids, flavonols, tannins, and phenols in all extracts with varying intensity. The organic extracts 100% methanol, 80% and 100% ethanol exhibited high color intensity (+++) for flavonoids and flavonols respectively, while all the extracts exhibited a moderate color intensity (++) for tannins and phenols. Spectrophotometric analysis of S. frutescens extracts revealed that all the organic extracts contained a significantly higher concentration (in mg/g of extract) of flavonols and tannins when compared to the aqueous extracts. All extracts contained approximately equal levels of phenols. These data confirm the presence of all four groups of bioactive phytocompounds in the S. frutescens extracts used in this study, and also confirm that different solvent extractants possess the capability to differentially extract specific groups of phytocompounds. in individual extracts. Further comparison of these compounds with online databases of anti-diabetic phytocompounds led to the preliminary identification of 10 possible anti-diabetic compounds; α-Pinene, Limonene, Sabinene, Carvone, Myricetin, Rutin, Stigmasterol, Emodin, Sarpagine and Hypoglycin B in crude and solid phase extraction (SPE) fractions of S. frutesecens. Furthermore, using two hepatic cell lines (Chang and HepG2) as an in-vtro model system, the anti-T2DM properties of crude aqueous and organic extracts of S. frutescents was investigated and compared. Both aqueous and organic extracts of S. frutescens were found to decrease gluconeogenesis, increase glucose uptake and decrease lipid accumulation (Triacylglycerol, Diacylglycerol, and Monoacylglycerol) in Chang and HepG2 hepatic cell cultures made insulin resistant (IR) following exposure to high concentration of insulin and fructose. Using real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the aqueous and organic extracts of S. frutescens were confirmed to regulate the expression of Vesicle-associated membrane protein 3 (VAMP3), Mitogen-activated protein kinase 8 (MAPK8), and Insulin receptor substrate 1 (IRS1) in insulin resistant hepatic cells. IR-mediated downregulation of VAMP3, MAPK8, and IRS1 mRNA in IR HepG2 hepatic cell cultures was reversed in the presence of aqueous and organic extracts of S. frutescens. The hot aqueous extract displayed the highest activity in all the assays, while all the organic extracts displayed similar potency. In conclusion, this study reports that aqueous and organic extracts of S. frutescens possess numerous anti-diabetic compounds that can be further investigated for the development of new, more efficacious and less toxic anti-diabetic agents. The presence of multiple compounds in a single extract does suggest a synergistic or combinatorial therapeutic effect. These findings support the burgeoning body of in-vivo and in-vitro literature evidence on the anti-diabetic properties of S. frutescens and its use in folklore medicine.

The Best M.Phil Thesis in the Faculties of Dentistry, Engineering, Medicine and Science (University of Hong Kong), Li Ka Shing Prize,1995-1997
published_or_final_version
Chemistry
Master
Master of Philosophy

Willow (Salix spp.) is a rich source of naturally occurring bioactive compounds. This study focussed specifically on exploring the condensed tannins (CT) in the National Willow Collection, which is a unique germplasm collection held at Rothamsted Research. Willow leaves were analysed by two different tannin methods, thiolysis and acid-butanol-acetone, in order to evaluate their suitability for assessing inter-accession variation and to identify accessions with a potential medicinal use. This is the first report of the tannin contents and compositions of a willow germplasm collection. Large variations were found for tannin contents, i.e. 2.8 to 19.5 g CT/100 g dry weight, and also for composition; mean degrees of polymerisation (mDP) ranged from 4.9 to 12.0, the procyanidin (PC)/prodelphinidin (PD) ratios from 19.1/80.9 to 52.0/48.0 and the cis/trans flavan-3-ol ratios from 13.0/87.0 to 52.1/47.9. Catechin was the major terminal unit and gallocatechin the major extension unit, which clearly distinguishes willow tannins from most other plant tannins. Interestingly, some accessions also had leaves with relatively unusual CT types, i.e. high mDP-values and high molar pcercentages of trans-flavan-3-ols. Cluster analysis succeeded in allocating several accessions into distinct groups that coincided with Salix breeding populations. This study has now laid the foundation for identifying the Salix tannin genome in the future. Given the time-comsuming nature of two tannin analysis methods. This study also evaluated the potential of Near-Infrared Spectroscopy (NIRS), which a much faster method, for predicting CT contents and compositions in these willow leaves. The results confirmed that NIRS is an excellent tool for estimating CT contents, PC/PD and c/s/frans-flavan-3-ol ratios in willow leaves. This finding could be of interest to plant breeders in developing new willow accessions. The final part of the thesis evaluated the antiparasitic potential of willow leaves against an intestinal parasite. CT with contrasting compositions were purified to test their efficacy against Haemonchus contortus, which is a major gastrointestinal parasite of livestock. As expected PD- were more active than PC-tannins. In addition, this study demonstrated for the first time that the PC activity could be synergistically enhanced by some Salix flavonoids.

This thesis investigated the pharmacology and phytochemistry of Achillea millefolium L. (yarrow) flowers, roots, stems, and leaves based on ethnobotanical reports in North America, with a focus on applications in a respiratory model. Seasonal changes in the phytochemical profile of yarrow were also assessed. A comprehensive dataset of medicinal Asteraceae was created after collecting ethnobotanical reports from the Native American Ethnobotany (NAEB) database. Using residual and binomial analyses, 14 tribes of Asteraceae were quantitatively evaluated and ranked within ten therapeutic categories as either over- or under-selected for treatment by North American indigenous peoples. Flora belonging to the Anthemideae tribe were over-utilized as pulmonary aids, particularly species of Achillea. Yarrow was selected for further analysis in the subsequent chapters of this thesis. The respiratory pharmacology of yarrow was examined by testing the immunomodulatory effects of four plant parts in an in vitro assay using BEAS-2B human bronchial epithelial cells. Concentrations of the pro-inflammatory cytokines IL-6 and IL-8 were quantified using ELISA kits. Flowers demonstrated significant anti-inflammatory activity at 40 μg/ml in both assays, and also at 20 μg/ml in the IL-8 assay, suggesting a dose-dependent response. Roots displayed significant pro-inflammatory activity at all concentrations. A second mechanism of action via the endocannabinoid system was tested through inhibitory enzyme assays for fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), in which the flowers and roots were most active. Since extracts of the four plant parts exhibited significantly different bioactivities, active metabolites previously identified in yarrow were quantified in each part through the targeted profiling of phenolics and alkylamides using analytical chromatographic techniques. Phenolic compounds were found at highest concentrations in the flowerheads, while alkylamides were detected predominantly within roots. An accompanying phenological analysis of alkylamide and phenolic levels in all parts was explored. Collectively, this research provides the first integrated comparison of yarrow ethnobotany, bioactivity, and phytochemistry across different parts of the plant, contributing novel insights into the traditional, contemporary, and future uses of one of North America’s most important medicinal plants.

PhD (Department of Biotechnology, Faculty of Applied and Computer Sciences), Vaal Universtiy of Technology
The genus Tulbaghia has been used in traditional medicine to treat various ailments such as fever, earache, tuberculosis and esophageal cancer. However, there is limited scientific evidence to support its use. Therefore the objectives of this study were to perform phytochemical analysis, investigate the antioxidant, antimicrobial, anticancer, immunomodulatory activities and toxicity of crude acetone and water extracts from selected Tulbaghia species. Standard methods were used for preliminary phytochemical analysis. The total phenolic content of the plant extracts was determined using the folin ciocalteu method whereas the total flavonoids were determined by using the aluminium chloride colorimetric method. DPPH and ABTS assays were used to evaluate the antioxidant activity. The antimicrobial activity was assessed by agar well diffusion, microtiter dilution and time kill assays. For anticancer studies, the antiproliferative activity of the extracts was evaluated using the MTT assay on Hkesc-1 and KB cells. Morphological changes of the cancer cells treated with extracts were examined using light microscopy. Induction of apoptosis was assessed using fluorescence microscopy and acridine orange/ethidium bromide staining. Flow cytometry analysis was conducted to examine the multicaspase activity and cell cycle arrest. For immunomodulatory activity, the Greiss reagent and Luminex cytokine assays were used to determine the effect of the extracts on NO production and the concentration of the cytokines in the treated cells, respectively. Toxicity of selected Tulbaghia species was examined by investigating the effect of the extracts on the metabolic activity and cell membrane integrity on the treated RAW264.7 cells using the MTT and LDH assays, respectively. The zebrafish assay was used to evaluate the embryotoxicity and teratogenic effects of crude acetone and water extracts of T. violacea at 24 h intervals for 96 h post fertilisation (hpf). The percentage mortality, hatchability and heart rate were examined. Phytochemical screening of eight Tulbaghia species demonstrated the presence of flavonoids, glycosides, tannins, terpenoids, saponins and steroids. The amount of total phenol and flavonoid content varied in different plant extracts ranging from 4.50 to 11.10 milligrams gallic acid equivalent per gram (mg GAE/g) of fresh material and 3.04 to 9.65 milligrams quercetin equivalent per gram (mg QE/g) of fresh material respectively. The IC50 values based on DPPH and ABTS for T. alliacea (0.06 and 0.06 mg/mL) and T. violacea (0.08 and 0.03 mg/mL) were generally lower showing potential antioxidant activities. For antimicrobial activity, the acetone extracts of T. acutiloba, T. alliacea, T. leucantha, T. ludwigiana, T. natalensis and T. simmleri showed moderate antimicrobial activity against all test organisms while the water extracts showed moderate to no activity. One species, T. cernua, showed poor activity against all the tested microbes. The acetone and water extracts of T. violacea showed the greatest antibacterial and antifungal activity against all the tested microorganisms with minimum inhibitory concentration ranging from 0.1 mg/mL to 3.13 mg/mL. The acetone extracts of T. violacea also exhibited both bacteriostatic/fungistatic and bactericidal/fungicidal activity depending on the incubation time and concentration of the extract. The bactericidal/fungicidal activity was observed at x2 MIC. The results for anticancer activity showed that treatment of Hkesc-1 cells with acetone and water crude extracts had anti-proliferative activity with IC50 values of 0.4 mg/mL and 1.625 mg/mL, respectively while KB had 0.2 mg/mL and 1 mg/mL, respectively. Morphological changes such as blebbing, cell shrinkage and rounding were observed in the treated cells suggesting that apoptosis was taking place. AOEB staining showed that the level of apoptosis was dependent on the concentration of the extracts. The activation of multicaspase activity in both Hkesc-1 and KB treated cells was also concentration dependent leading to cell death by apoptosis and the induction of cell cycle arrest at the G2/M phase. Immunomodulatory activity results indicated that cell viability was above 80% when concentrations of 50 µg/mL or less of both acetone and water crude was used. Treatment with the acetone extract had no significant effect (p>0.05) on the LPS induced NO production in RAW264.7 cells except at 50 µg/mL where significant inhibition was observed. The water extract had no significant effect (p>0.05) on NO production at all the concentrations. Treatment of LPS–induced RAW264.7 cells with acetone extract stimulated the production of IL-1α, IL-6 and TNF-α, but had no significant effect (p > 0.05) on IL-1β. On the other hand, treatment with the water extracts stimulated the production of IL-1α, IL-6 but had no significant effect (p>0.05) on TNF-α and IL-1β. Treatment of LPS-induced RAW264.7 cells with the acetone extract had very little stimulatory effect on IL-4, IL-5 and IL-13 and no significant effect on IL-10 whereas for the water extract a significant stimulatory effect was only observed for IL-4 after 48 h of treatment. High concentrations (>10000 pg/mL) of MCP-1, MIP1-α, MIP1-β, MIP-2, GCSF, GM-CSF, RANTES and IP-10 were also observed in acetone and water extract treated RAW264.7 cells. For toxicity studies, acetone and aqueous crude leaf extracts from T. alliacea, T. simmleri, and T. violacea had a significant inhibitory (p<0.05) effect on the RAW264.7 cells after 48h treatment. Acetone extracts from T. alliacea, T. simmleri and T. violacea resulted in IC50 values of 0.48 mg/mL, 0.72 mg/mL and 0.1 mg/mL, respectively. Treatment with water extracts showed minimal toxic effect indicated by higher IC50 values of 0.95 mg/mL, 2.49 mg/mL and 0.3 mg/mL for T. alliacea, T. simmleri and T. violacea, respectively. The LDH release by macrophages after 24 h treatment with acetone extracts was observed to be concentration dependent while treatment with water extracts did not induce LDH release. The zebra fish assay showed a lethal dose (LD50) for the T. violacea acetone crude extract of 20 μg/mL whereas that for water extract was 85 μg/mL. The observed teratogenic effects included scoliosis, edema of the pericardial cavity, retarded yolk resorption, hook-like/bent tail and shorter body length. In conclusion, the results from this study indicate that the extracts from the eight Tulbaghia species examined contain phytochemicals that may have the antioxidant, antimicrobial, anticancer and immunomodulatory properties. Extracts from T. violacea were observed to be the most potent. This study thus supports the use of T. violacea in treating bacterial and fungal infections in traditional medicine. The results of this study also confirm the anticancer potential of T. violacea. The immunomodulatory activity of the acetone and water extracts from T. violacea indicated a dominantly pro-inflammatory activity. Traditional medicine prepared form T. violacea may be of benefit to individuals with weak immune systems. The toxicity of selected Tulbaghia species was observed to be concentration, extract and time dependent. Therefore, traditional medicine prepared from Tulbaghia extracts should be taken with caution preferably in small doses over a short period of time. Future studies will focus on the identification of the bioactive compound(s) responsible for the antimicrobial, anticancer and immunomodulatory activities.

Abstract Medicinal plants have been for long remedies for human diseases because they contain components of therapeutic value. The growing problem of antibiotic resistance by organisms demands the search for novel compounds from plant based sources. The present study was aimed at evaluating the bioactivity and phytochemical analysis of Hydnora africana on clinical and standard strains of Helicobacter pylori (PE 252C and ATCC 43526), Aeromonas hydrophila ATCC 35654, and Staphylococcus aureus NCT 6571 in an effort to identify potential sources of cheap starting materials for the synthesis of new drugs against these strains. Ethyl acetate, acetone, ethanol, methanol, and water crude extracts of H. africana were screened for activity against the test organisms using the agar well diffusion assay. The Minimum Inhibitory Concentration (MIC50) and Minimum Bactericidal Concentration (MBC) of the most potent extracts were determined by the microdilution method, followed by qualitative phytochemical analysis. Results were analyzed statistically by ANOVA one - way test. Different concentrations (200,100, 50mg/mL) of the methanol, acetone, ethanol and ethyl acetate extracts showed activity against S. aureus and A. hydrophila while for H. pylori, only methanol and ethyl acetate extracts were active; water showed no activity for all studied bacterial pathogens. Mean zone diameter of inhibition which ranged from 0-22mm were observed for all test bacterial pathogens and 14-17mm for ciprofloxacin. The activity of methanol and ethyl acetate extracts were statistically significant (P< 0.05) compared to all the other extracts. MIC50 and MBC ranged from 0.078 – 2.5mg/mL, 0.78-25mg/mL respectively for all tested bacterial pathogens. For ciprofloxacin, the MIC50 and MBC ranged from 0.00976 – 0.078mg/mL and 0.098– 0.78mg/mL respectively. There was no statistically significant difference between extracts (methanol, acetone, ethanol, ethyl acetate) and the control antibiotic (ciprofloxacin) (P> 0.05). Qualitative phytochemical analysis confirmed the presence of alkaloids, saponins, steroids, tannins and flavonoids in the methanol, acetone,ethanol and ethyl acetate extracts. The results demonstrate that H. africana may contain compounds with therapeutic potentials which can be lead molecules for semi-synthesis of new drugs.

Echinacea purpurea is consumed as a natural health product around the world. Due to the genus’ ethnobotanical relevance, the phytochemistry of Echinacea has been extensively studied, revealing a variety of bioactive metabolites including caffeic acid derivatives and alkylamides. Whereas seasonal trends in root chemistry have been established, trends in other plant parts are relatively understudied. Similarly, few studies have evaluated the effects of organic plant growth substances in field trials. With increased demand for organic products, industry is looking for alternative ways to optimize yields and medicinal properties. For this thesis, my first objective was to quantify the concentrations of E. purpurea’s secondary metabolites across organic treatments throughout the plant’s first growth year to determine optimal harvesting time and conditions in all parts of the plant. The second objective was to determine how seasonal variations affect its potential bioactivity through inhibition of Pseudomonas aeruginosa. Plants were grown in field plots treated with four different organic treatments: water (control), high cytokinin, low cytokinin, and fish oils; samples were collected biweekly from May-September. Dried plants were separated into major plant parts and were extracted exhaustively in 70% EtOH. Using high-pressure liquid chromatography (HPLC), concentrations of alkylamides and select caffeic acid derivatives were quantified in all samples and compared across plant part, developmental stage, and organic fertilizers. It was determined that while there were no major differences between treatments, phytochemical concentrations changed throughout the season in all plant parts; revealing that aerial parts of the plant also bioactive secondary metabolites and should not be excluded from future studies. Following this study, an MIC50 assay was used to test these extracts against P. aeruginosa PA14. It was noted that seasonality effects of phytochemistry were not consistent with bioactivity and that there were no significant differences between extract and carrier control. While the antibiotic activity of root extracts varied seasonally, the flower extract exhibited the most consistent antibiotic potential. The results presented in this thesis will not only aid in industry practices and yield optimization but, through filling knowledge gaps on seasonality and organic treatments in field trials, will increase the understanding of E. purpurea’s chemistry and related bioactivity, with implications on both the medicinal properties and eco-physiology of the species.

Cancer and malaria are among the most life-threatening diseases globally. Cancer is responsible for about 125,000 annual deaths globally. In 2015, the World Health Organization report estimated that 236000-635000 people died of malaria. These diseases are complicated by the development of resistance to available chemotherapeutic agents. Natural products have been recognized for their major applications in the identification of drug leads in drug discovery. Viola philippica Car, Viola yedoensis Makino (Violaceae), Triclisia subcordata Oliv (Menispermeaceae) and Cyclicodiscus gabunensis Harms (Fabaceae) are medicinal plants traditionally used for the treatment of various diseases including malaria or cancer in China and West Africa. However, the bioactive compounds are unknown. Therefore, this study evaluated the in vitro anticancer and antimalarial activities of the four medicinal plants and searched their bioactive compounds. The in vitro anti-ovarian cancer and antimalarial assays were demonstrated respectively using sulforhodamine B dye and Syber green 1 fluorescence assay methods. Bioassay-guided fractionation and purification were performed. Structural elucidation was performed by nuclear magnetic resonance spectroscopy and mass spectrometry analysis. Results revealed the anticancer and antimalarial activities of T. subcordata; V. philippica, and V. yedoensis to be bisbenzylisoquinoline alkaloids (cycleanine, isochondodendrine and 2′-norcocsuline) and/or cyclotides. The cycleanine analogues were synthesized and found to be more potent than cycleanine. Induction of apoptosis by these alkaloids has also been determined. This study could serve as basis for the support of use of these plants in cancer and/or malaria treatment. The BBIQ alkaloids and analogues could serve as lead compounds in drug discovery. Future in vivo studies need to be carried out on these alkaloids to get drug approval.

Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.

Orientador: Rosemeire Cristina Linhari Rodrigues Pietro
Banca: Rosemeire Cristina Linhari Rodrigues Pietro
Banca: Luis Carlos Marques
Banca: Taís Maria Bauab
Resumo: Embora milhares de espécies vegetais sejam utilizadas na medicina tradicional do mundo, estima-se que apenas 1% são conhecidas por estudos científicos, com valor terapêutico demonstrado. As espécies Polygonum acre e Synadenium carinatum são de fácil cultivo, estando largamente distribuídas no Brasil, e possuindo um alto índice de utilização na medicina popular. Entre os usos populares da espécie Polygonum acre podem ser citados: anti-séptico, antiinflamatório, hipotensor, anti-hemorroidal, diurético, vermicida e anti-diarréico. A espécie Synadenium carinatum é popularmente utilizada para o tratamento de cânceres. No entanto, não existem estudos científicos que comprovem esses efeitos, nem informações sobre a segurança de utilização dessas drogas pelos seres humanos além de haver poucos estudos fitoquímicos destas espécies. Neste trabalho foram realizados os estudos fitoquímico, microbiológico, a busca de atividades biológicas dos extratos dessas plantas e a investigação de aspectos relativos à segurança de utilização. Os testes fitoquímicos indicaram a presença de flavonóides, taninos, saponinas, mono, sesqui e diterpenos, e derivados cinâmicos nas espécies P. acre e S. carinatum, enquanto que apenas a espécie P. acre respondeu positivamente quanto à presença de proantocianidinas condensadas e leucoantocianidinas. Na avaliação da atividade antimicrobiana através do método de difusão em ágar e da técnica de Concentração Inibitória Mínima, a espécie P. acre apresentou atividade antimicrobiana nas concentrações de 300 mg/mL, enquanto a atividade antimicrobiana para os extratos de S. carinatum não foi evidenciada. Nos testes de toxicidade aguda, pode-se observar que na dose de 2 g/kg, o extrato etanólico 70% de P. acre apresentou taxa de mortalidade de 50%, enquanto o extrato etanólico 70% de S. carinatum não apresentou toxicidade ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Although there are thousands of plant species used in traditional medicines in the world, it is estimated that only 1% are known by scientific studies with demonstrated therapeutic value. The species Polygonum acre and Synadenium carinatum are easy to culture, being widely distributed in Brazil, and having a high rate of use in folk medicine. Among the popular uses of the species Polygonum acre may be cited anti-septic, anti-inflammatory, hypotensive, anti-hemorrhoid, diuretic, vermifuge, and anti-diarrheal. The Synadenium carinatum species is popularly used in the treatment against cancer. However, there are no scientific studies that show these effects, no information about the safe use of these drugs by human beings and there are few phytochemical studies of this species. In this work, the phytochemical study and microbiological quality control, the search for biological activities of the extracts of plants and the investigation of aspects related to safe use were done. The phytochemical tests indicated the presence of flavonoids, tannins, saponins, mono, sesqui and diterpenes, and cinnamic derivatives in P. acre and S. carinatum species, while only the species P. acre responded positively about the presence of condensed proanthocyanidins and leucoanthocianidins. In the evaluation of antimicrobial activity by the ágar diffusion method and Minimum Inhibitory Concentration technique, the P. acre species had antimicrobial activity at concentrations of 300 mg / mL, while the antimicrobial activity for the extracts of S. carinatum was not demonstrated. In the tests of acute toxicity the ethanol extract 70% of P. acre at a dose of 2 g/kg showed 50% of mortality while the ethanol extract 70% of S. carinatum has no toxicity. At the dose of 1 g/kg the 70% ethanol extract of P. acre showed 50% of mortality while the latex of S. carinatum caused 100% of death the mice... (Complete abstract click electronic access below)
Mestre

Syzygium cordatum Hoscht ex. C Krauss, also known as water berry, is normally used by the people of South Africa for respiratory ailments including tuberculosis, stomach complaints, treatment of wounds and as emetics. An extract of the leaves can be used as a purgative for diarrhoea treatment. The leaves of Syzygium cordatum Myrtaceae were obtained from the Eastern Cape Province of South Africa, air dried and sequential solvent extraction was done to obtain various non volatile crude extracts. The volatile extract, that is the essential oil was extracted from the leaves using hydrodistillation and analysis of compounds was done by GC/MS for composition. 32 compounds were obtained from the fresh leaves and 18 compounds were obtained from the dry leaves. The fresh oil contains caryophyllene (11.8 percent) and caryophyllene oxide (11.1 percent) as the main sesquiterpene component. α-Pinene(5.0 percent) was the only monoterpene compound identified in the fresh oil in substantial amount. The dry leaves oil had copanene (17.0 percent), β-Caryophellene (26.0 percent), cubenol (6.5 percent) and caryophellene oxide (14.2 percent) as the dominant constituent of the oil. Summary of the classes of compounds in the oil revealed that the chemical profile of both oils were dominated by sesquiterpenoid compounds. This is the first time that terpenoids compounds are being identified in both the fresh and dry leaf oil of S. cordatum. Hexane leaf extract was selected due to the interest in the terpenoid compounds. Column chromatography of the hexane crude gave five (5) of which two are fully reported. The isolates were fully elucidated using spectroscopic methods to be β-Sitosterol (HC3) and Friedela-3-one (HC1A/HC1D). Cytotoxicity analysis was carried out on the crude using the Brine shrimps assay. Isolates 1C and1D showed significant lethality using the brine shrimps assay with lethality values (LC50) of 4.105mg/ml for HC1C and 4.11mg/ml for 1D/1A respectively.

The first study was looking into the phytochemical analysis of herbal medicinal plant from Malaysia. Phenolic compounds in an aqueous infusion from leaves of Ficus deltoidea (Moraceae), a well-known herbal tea in Malaysia, were analysed by HPLC coupled to photodiode-array and fluorescence detectors and an electrospray ionization tandem mass spectrometer. Following chromatography of extracts on a reverse phase C12 column 25 flavonoids were characterized and/or tentatively identified, with the main constituents being flavan-3-ol monomers, proanthocyanidins and C-linked flavone glycosides. The proanthocyanidins were dimers and trimers comprising (epi)catechin and (epi)afzelechin units. No higher molecular weight proanthocyanidin polymers were detected. The antioxidant activity of F. deltoidea extract was analysed using HPLC with on-line antioxidant detection. This revealed that 85% of the total antioxidant activity of the aqueous F. deltoidea infusion was attributable to the flavan-3-ol monomers and the proanthocyanidins In terms of the bioavailability of hydroxycinnamates with the high occurrence of p-coumaroylquinic acid in F. deltoidea extract, further study was carried out using radiolabeled caffeic acid. In the study, male Sprague-Dawley rats ingested 140 x 106 dpm of [3-14C]trans-caffeic acid and over the ensuing 72 h period body tissues, plasma, urine and faeces were collected and the overall levels of radioactivity determined. Where sufficient radioactivity had accumulated samples were analysed by HPLC with on-line radioactivity and tandem mass spectrometric detection. Nine labeled compounds were identified, the substrate and its cis isomer, 3¢-O- and 4¢-O-sulphates and glucuronides of caffeic acid, 4¢-O-sulphates and glucuronides of ferulic acid and isoferulic acid-4¢-O-sulphate. Four unidentified metabolites were also detected. After passing down the gastrointestinal tract the majority of the radiolabeled metabolites were excreted in urine with minimal accumulation in plasma. Only relatively small amounts of unidentified 14C-labeled metabolites were expelled in faeces. There was little or no accumulation of radioactivity in body tissues, including the brain. The overall recovery of radioactivity 72 h after ingestion of [3-14C]caffeic acid was ~80% of intake. The role of colonic microflora in the metabolism of caffeic acid was carried using an in vitro model of the human colonic microflora. Caffeic acid (55 µmoles) was incubated with human faecal materials obtained from five Asian donors, and caffeic acid degradation was monitored from 0-8 h. Faecal samples were analysed by GC-MS, where major phenolic acids identified were dihydrocaffeic acid, 3-(3¢-hydroxyphenyl)propionic acid and phenylacetic acid. Caffeic acid was quickly degraded by the colonic microflora, as it disappeared after 4 h of incubation in two of the faecal samples. The degree of degradation of caffeic acid was significantly influenced by the addition of glucose as well as individual variations in the density and the composition of microflora. These findings support extensive metabolism of caffeic acid in the colon, depending on the substrate concentration and the supplement of glucose which resulting the formation of simple phenolics.

Magister Pharmaceuticae - MPharm
Evaluation of the antinociceptive, anti-inflammatory and antipyretic activities of Ruta graveolens L. in mice and rats FIRDOWS LOONAT M. Pharm. Pharmaceutical Sciences thesis: School of Pharmacy, University of the Western Cape Ruta graveolens (Rutaceae) L. is a medicinal plant that is commonly used to manage and treat essential events such as pain, inflammation and fever. Despite its popularity, particularly as a medicinal plant in the Calvinia district and Bredasdorp region of South Africa, scientific data to substantiate its widespread traditional use and the possible mechanisms of action for this plant species is lacking. Therefore, the objectives of this study were: to scientifically evaluate and validate the antinociceptive, anti-inflammatory and antipyretic activities of Ruta graveolens using the acetic-acid writhing test and hot-plate test, the carrageenan rat paw oedema test, and the E. coli-induced pyrexia test, respectively; to investigate the possible mechanisms of the antinociceptive, anti-inflammatory and antipyretic activities of the plant using interaction studies; to determine some secondary metabolites present in the plant species using standard phytochemical analytical procedures; to characterise the plant species using HPLC techniques; and to determine the safety profile of the plant species using an acute toxicity study.Three percent (3 %) acetic acid (0.25 ml, i.p.) produced a substantial number of writhes in mice. The leaf methanol extract of Ruta graveolens (100 mg/kg, i.p.) significantly reduced the number of writhes induced by 3 % acetic acid (0.25 ml, i.p.). R. graveolens (100 mg/kg,i.p.) produced 54 % inhibition of 3 % acetic acid-induced writhes. Indomethacin (20 mg/kg,i.p.) and paracetamol (500 mg/kg, i.p.) significantly reduced the number of 3 % acetic acidinduced writhes. Indomethacin (20 mg/kg, i.p.) and paracetamol (500 mg/kg, i.p.) produced 57 % and 80 % inhibition of 3 % acetic acid-induced writhes, respectively. R. graveolens (25– 50 mg/kg, i.p. and 200 – 400 mg/kg, i.p.) and indomethacin (10 mg/kg, i.p.) did not significantly reduce the number of writhes induced by 3 % acetic acid. However, combined therapy of the leaf methanol extract of R. graveolens (25 mg/kg, i.p.) and indomethacin (10 mg/kg, i.p.) significantly reduced the number of 3 % acetic acid-induced writhes. The combined therapy of the lowest and sub-effective doses of the leaf methanol extract of R. graveolens (25 mg/kg, i.p.) and indomethacin (10 mg/kg, i.p.) produced 59 % inhibition of the writhes elicited by 3 % acetic acid. The leaf methanol extract of R. graveolens (50 – 400 mg/kg, i.p.) greatly delayed the reaction time in mice to thermal stimulation produced with hot-plate. 50 – 400 mg/kg (i.p.) of the leaf methanol extract of R. graveolens significantly antagonised rat paw oedema induced by 1 % carrageenan (0.1 ml, subplantar) over the 4 h period of testing. In addition, indomethacin (10 mg/kg, i.p.) significantly antagonised 1 % carrageenan-induced rat paw oedema. R. graveolens (25 mg/kg, i.p.) and indomethacin (2mg/kg, i.p.) given separately did not significantly alter rat paw oedema induced by 1 % carrageenan. However, combined therapy of the leaf methanol extract of R. graveolens (25 mg/kg, i.p.) and indomethacin (2 mg/kg, i.p.) significantly reduced 1 % carrageenan-induced rat paw oedema. The leaf methanol extract of R. graveolens (400 mg/kg, i.p.) significantly reduced the mean rectal temperature of normothermic rats. Ruta graveolens (100 – 400 mg/kg, i.p.) significantly reduced pyrexia induced by E. coli (50 μg/kg, i.m.) over the 5 h period of testing. In addition, pentoxifylline (50 mg/kg, i.p.) significantly reduced E. coliinduced pyrexia. Ruta graveolens (25 – 50 mg/kg, i.p.), paracetamol (500 mg/kg, i.p.) and pentoxifylline (10 mg/kg, i.p.) did not significantly reduce pyrexia induced by E. coli.However, combined therapy of the leaf methanol extract of R. graveolens (25 mg/kg, i.p.)and pentoxifylline (10 mg/kg, i.p.) significantly reduced E. coli (50 μg/kg, i.m.)-induced pyrexia.The phytochemical studies of the powdered leaves of Ruta graveolens indicated the presence of alkaloids, cardiac glycosides, flavonoids, saponins, tannins and triterpene steroids. The HPLC fingerprint indicated characteristic peaks at the following retention times; 1.654 min,2.271 min, 2.403 min, 4.705 min and 7.691 min. The LD50 obtained for Ruta graveolens after oral administration was probably greater than 4000 mg/kg which shows that the plant extract is non-toxic to mice.In conclusion, the data obtained indicate that Ruta graveolens possesses antinociceptive, antiinflammatory and antipyretic activities. Since prostaglandins have been shown to mediate acetic acid-induced writhes, prostaglandins, histamine, serotonin, capsaicin and bradykinin implicated in carrageenan-induced rat paw oedema, and tumor necrosis factor-α (TNF-α) implicated in E.coli-induced pyrexia, it is possible that R. graveolens may be producing its antinociceptive, anti-inflammatory and antipyretic activities by affecting these chemical mediators. The data obtained also justify the use of the plant species by traditional medicine practitioners for the treatment of painful and inflammatory conditions, and pyrexia.

Antimicrobial resistance is a worldwide problem that has deleterious long-term effects as the development of drug resistance outpaces the development of new drugs. Plants have been used for many generations for healing purposes, and screening of extracts of these plants has often yielded positive outcomes. This study was aimed at isolating and characterizing the major active antimicrobial compounds present in the stem bark of C. molle, in a bid to identify potential sources of cheap starting materials for the synthesis of new drugs. Various solvents (hexane, ethyl acetate, dichloromethane, acetone, ethanol and methanol) were used for extraction. The agar well diffusion technique was used to screen for antimicrobial activity of C. molle extracts against Streptococcus pyogenes ATCC 49399, Plesiomonas shigelloides ATCC 51903, Pseudomonas aeruginosa ATCC 15442, Helicobacter pylori ATCC 43526 and Helicobacter pylori 252C (clinical isolate); minimum inhibition concentration (MIC) of the most active extracts was determined by the broth dilution method. Fractionation of acetone extract was done by thin layer chromatography (TLC) and bioautography to determine the compounds present and their antimicrobial activity respectively. The acetone extract was purified by column chromatography and their MIC determined. The most potent fraction (EA4) was subjected to Gas chromatography- Mass spectrometry (GC-MS) and High performance liquid chromatography (HPLC) for identification of the active compounds. Results were analyzed by the Fisher‟s exact test. All the extracts tested demonstrated antimicrobial activity with zone diameters of inhibition ranging from 0–32 mm. Acetone was the most potent extract with its MIC ranging from 0.078–5.0 mg/mL. Seventeen fractions were collected from column chromatography and the most active fraction against all the organisms was EA 4 (eluted with 100 percent ethyl acetate), with its MIC ranging from 0.078 - 2.5mg/mL. There was no statistically significant difference (P>0.05) in the potency of the xii four extracts (acetone, methanol, ethanol and ethyl acetate) and antibiotic (ciprofloxacin) on the different bacterial strains tested, likewise the crude extract and the fractions. No compound was detected by GC-MS whereas numerous peaks were identified by HPLC implying that the active compounds in this plant are non volatile. We could not identify the compounds thereby proposing further studies using Nuclear magnetic resonance to identify the compounds. The study revealed that the acetone extract of C. molle was the most active against all the test organisms and therefore justifies the use of this plant in traditional medicine.

Medicinal plants have been used as traditional medicine in the treatment of numerous human diseases for thousands of years in many parts of the world. In the developing world, especially in rural areas, herbal remedies continue to be a primary source of medicine. Scientifically, medicinal plants have proven to be an abundant source of biologically active compounds, many of which have already been formulated into useful therapeutic substances or have provided a basis for the development of new lead molecules for pharmaceuticals. Antibiotic resistance, undesireable side effects and expences associated with the use of combination therapy in the treatment of Helicobacter pylori infections have generated a considerable interest in the study of medicinal plants as potential sources of new drugs against this organism. The high complexicity of bioactive compounds accumulated in plants coupled with their broad antimicrobial activity may make it difficult for pathogenic organisms, including H. pylori to acquire resistance during treatment. This study therefore evaluates the antimicrobial potential of selected South African medicinal plants employed in the treatment of H. pylori-related infections, and the subsequent isolation of the plant active principles. An ethnobotanical survey of plants used in the treatment of H. pylori-related infections was conducted in the study area. Crude extracts of Combretum molle, Sclerocarya birrea, Garcinia kola, Alepidea amatymbica and 2 Strychnos species were screened against 30 clinical strains of H. pylori and 2 standard control strains (NCTC 11638 and ATCC 43526). In the preliminary stages of this study, ethyl acetate, acetone, ethanol, methanol and water extracts of the plants were tested against H. pylori by agar well diffusion and micro broth dilution methods. The plant crude extracts that exhibited anti-H. pylori activity with a iv percentage susceptibility of 50 percent and above were considered for the rate of kill assays and the most active crude extracts selected for bio-assay guided isolation of the active ingredient. Preliminary fractionation of the crude extract was achieved by thin layer chromatography (TLC) using different solvent combinations; hexane/diethylether (HDE), ethyl acetate/methanol/water (EMW) and chloroform/ethyl acetate/formic acid (CEF) in order to determine the most suitable combination for column chromatography (CC) and subsequent testing by indirect bioautography. The extract was then fractionated in a silica gel column using previously determined solvent combinations as eluent. Active fractions obtained from column chromatography separations were further fractionated and the compounds identified by gas chromatography/mass spectrometry (GC/MS) analysis. All the plants exhibited antimicrobial activity against H. pylori with zone of inhibition diameters ranging from 0 - 38 mm and minimum inhibitory concentration (MIC) values ranging from 0.06 - 5.0 mg/mL. The most active plant extracts were the acetone extract of C. molle with a percentage susceptibility of 87.1 percent, acetone and aqueous extracts of S. birrea (71 percent each) and the ethanolic extracts of G. kola (53.3 percent). Except for the aqueous extract, these extracts also exhibited a strong bactericidal activity against H. pylori at different concentrations. TLC analysis revealed the presence of 9 components in the acetone extract of S. birrea with the EMW solvent system as opposed to 5 and 8 with HDE and CEF respectively. Bioassay-guided isolation led to the identification of 52 compounds from the acetone extract of S. birrea with n-octacosane being the most abundant (41.68 percent). This was followed by pyrrolidine (38.91 percent), terpinen-4-ol (38.3 percent), n-eicosane (24.98 percent), cyclopentane (16.76 percent), n-triacontane (16.28 percent), aromadendrene (13.63 percent) and α-gujunene (8.77 percent). Terpinen-4-ol and pyrrolidine demonstrated strong antimicrobial activity against H. pylori at all concentrations tested. These results may serve as preliminary scientific validation of the ethnomedicinal uses of the above mentioned plants in the treatment of H. pylori-related infections in South Africa. Terpinen-4-ol and pyrrolidine could be considered for further evaluation as therapeutic or prophylactic agents in the treatment of H. pylori-related infections. However, further investigations would be necessary to determine their toxicological properties, in-vivo potencies and mechanism of action against H.pylori

Diverse pharmacological activities and the solid clinical performance of Chinese herbal medicines have attracted worldwide attention in terms of its modernization. Here, a hexaherbal Chinese formula (HHCF) for treating atopic dermatitis (AD) topically has been studied from chemical and biological perspectives. The HHCF comprises the rootstock of Scutellaria baicalensis Georgi (SCU), Rheum tanguticum Maxim. Ex Balf. (RHE), Sophora flavescens Aiton (SOP); the root bark of Dictamnus dasycarpus Turcz. (DIC); the bark of Phellodendron chinense C.K. Schneid. (PHE) and the fruit of Kochia scoparia (L.) Schrad. (KOC). In this thesis the chemical composition of the HHCF has been profiled and characterized using liquid-chromatography (LC) coupled with triple quadruple mass spectrometry (MS). 68 chemical compounds including alkaloids, anthraquinones, coumarins, flavonoids, naphthalene derivatives, phenylbutanone glucosides, phenolic acids, pterocarpans, stilbenes and tannins were putatively identified in the LC-MS profile of the HHCF based on mass measurement and characteristic fragment ions. The source(s) of these chemical compounds has been analyzed using the developed EXCEL template and PHE, RHE and SOP contributed the largest number of chemical compounds identified in the HHCF, while KOC seems to contribute very little. To evaluate the anti-inflammatory effects of the HHCF for treating AD/skin inflammation, the thymus and activation-regulated chemokine (CCL17), Prostaglandin E2 (PGE2), IL-8, IL-6 and hyaluronidase inhibitory effects were studies in vitro. Results showed that the HHCF inhibited hyaluronidase activity, TNF-α- plus IFN--induced CCL17 production in spontaneously immortalized human epidermal keratinocytes (HaCaT) and IL-1β-induced PGE2 in human fibroblasts with no effects on IL-8 and IL- 6. Among the chemical compounds characterized in LC-MS profile of the HHCF, multivariate regression models indicated the major contributor to the CCL17 inhibition were berberine, catechin dimers, gallic acid, galloylglucose, 4-(4'-hydroxylphenyl)-2- butanone 4'-O-β-D-glucoside, pyrogallol and resveratrol 4'-O-β-D-(6''-O-galloyl) glucoside. The effects of the HHCF against other pathogenesis-related targets need to be further study to ascertain its therapeutic efficacy against AD.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico
O presente trabalho relata o estudo dos constituintes quÃmicos volÃteis das folhas, talos e raÃzes de Croton rhamnifolius (Euphorbiaceae), popularmente conhecida no nordeste do Brasil por âQuebra-facaâ, como tambÃm os constituintes quÃmicos nÃo-volÃteis das raÃzes. A anÃlise por CGL/EM do Ãleo essencial das folhas de C. rhamnifolius (OEFCR) permitiu a identificaÃÃo de 21 constituintes quÃmicos, dos quais os componentes majoritÃrios foram identificados como sendo o 1,8 â cineol (31,81%) e o â felandreno (11,84 %). O Ãleo essencial dos talos (OETCR) apresentou 14 constituintes, todos identificados, onde o -pineno (30,81%) e o âcariofileno (12,86 %) foram os componentes principais. No Ãleo essencial das raÃzes (OERCR), foram identificados 8 constituintes quÃmicos, sendo majoritÃrios, o valenceno (25,97%), e o borneol (20,05%). A anÃlise fitoquÃmica do extrato etanÃlico da raiz resultou no isolamento de um alcalÃide indÃlico diterpenoÃdico de carÃter inÃdito que foi denominado rhamnifolina (1), dois diterpenos do tipo beiereno, denominado de estachenona (2) e 12-oxo-estachenona (3), e de dois triterpenos pentacÃclicos, um da sÃrie oleanano, o Ãcido 3-acetil-olean-14-en-28-Ãico (Ãcido acetil aleuritÃlico, AAA) (4) e o lupeol (5) da sÃrie lupano. A determinaÃÃo estrutural dos constituintes nÃo volÃteis foi realizada atravÃs de mÃtodos espectromÃtricos EM, IV e RMN 1H e 13C, inclusive tÃcnicas uni e bidimensionais (HMBC e HSQC), enquanto a identificaÃÃo da constituiÃÃo volÃtil foi utilizada cromatografia gÃs-lÃquido acoplada à espectrometria de massa (CGL/EM) e cromatografia gÃs-lÃquido com detecÃÃo por ionizaÃÃo em chama (CGL/DIC) e comparaÃÃo com dados descritos na literatura.
The present work reports the study on the volatile chemical constitution of the leaves, twigs and roots of Croton rhamnifolius (Euphorbiaceae), popularly known in the Northeast of Brazil as âquebra-facaâ (Port. Lit. = knife breaker) as well as of the non-volatile constituints of the roots. GC-MS analysis of the essential oil from leaves of C. rhamnifolius (OEFCR) allowed the identification of 21 constituents from which 1,8-cineol (31,81%) and - phelandrene (11,84%) were the major components. For the essential oil from twigs (OETCR) all 14 components were identified and - pinene (30,81%) and - cariophyllene (12,86%) were the major ones. For the essential oil from roots (OERCR) jsut 8 components were identified with valencene (25,97%) and borneol (20,05%) arising as the chief components. The phytochemical analysis of the ethanol extract from the roots led to the isolations and characterization of a diterpenoid incorporating an indol alkaloid moiety that hÃs been named as rhamnifolin (1), two beierene diterpenes, stachenone (2) and 12-oxostachenone (3), and two pentacyclic triterpenes: one, the 3-acetyl-olean-14-ene-28-oic acid (aleuritolic acid, AAA) (4) belonging to the oleanane and the other, lupeol (5), belonging to the lupane series. The structure determination of all non-volatile constituents was performed by mean of spectroscopic techniques such as MS, IR, 1H and 13C NMR, including uni and bidimensional pulse sequences. The identification of the volatile constitution was performed by GC-MS for the qualitative analysis while GC-FID was used for the quantitative analysis. Comparison to data published in the literature was also used for identification wherever the case.

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CAPES
Para tornar potáveis águas salobras ou salinas, é necessário fazer a dessalinização, processo que normalmente exige alto investimento e recursos tecnológicos complexos para a produção em larga escala. Uma solução simples e eficiente para as comunidades do Semi Árido pode ser a dessalinização de água salobra pelo contato com materiais biológicos, através do uso de dispositivos caseiros contendo, por exemplo, sementes que adsorvam parte do sal da água, reduzindo a salinidade da mesma. Em estudo desenvolvido recentemente sobre a capacidade de sorção de sais de água salobra por sementes típicas de regiões semi áridas foram consideradas mais eficientes sementes de umbu (Spondias tuberosa Arruda) e Umburana (Amburana cearensis A. C. Smith). Em função disso, foi estudado neste trabalho o perfil químico desses materiais biológicos no seu estado natural e transformado em carvão quando aquecido a 250 °C por 1 hora, que foi a melhor condição encontrada para maior eficiência de dessalinização. Da análise fitoquímica da semente da umburana foi observada a presença de alcalóides, taninos, saponinas, cumarinas, triterpenos, esteróides e flavonóides, e o seu carvão apresentou resultados positivos para alcalóides, taninos, saponinas, esteróides e flavonóides. Na semente do umbu observou-se a presença de taninos, cumarinas, ácidos orgânicos, esteróides e flavonóides e no carvão não foram observados resultados positivos. No perfil dos ácidos graxos obtido por cromatografia gasosa (CG) dos ésteres metílicos produzidos por reação de esterificação dos extratos hexânicos das sementes, foram encontrados: na semente in natura da umburana dez ácidos graxos, sendo oito saturados (palmitíco, margárico, oléico, esteárico, nnonadecílico, araquídico, n-heneicosóico e lignocérico), um insaturado (ácido oléico) e outro que não foi possível identificar; o carvão desta semente apresentou os mesmos ácidos graxos, além do ácido mirístico; a semente in natura do umbu apresentou sete ácidos graxos saturados (mirístico, palmitíco, esteárico, nnonadecílico, araquídico, n-heneicosóico e behênico) e dois ácidos insaturados (oléico e linoléico); o seu carvão, apresentou a mesma composição dos ácidos graxos saturados que a semente in natura, mas apenas um insaturado (ácido oléico). A análise por CLAE dos extratos da semente da umburana in natura e como carvão e do umbu in natura revelou pela primeira vez à presença de ácido Lascórbico e ácido gálico nestes materiais biológicos. A presença de alcalóides identificados no carvão da umburana não é um bom indicativo para sua utilização como dessalinizador, devido à conhecida toxicidade de substâncias dessa família, assim como pela presença de cumarina, em função da sua toxicidade e solubilidade parcial em água. No entanto, devido à facilidade de obtenção da cumarina a partir da semente moída da umburana, sua utilização poderá ser viável em indústrias de produtos de limpeza e cosméticos ou como material de partida para síntese de fármacos, entre outros. No carvão produzido com a semente de umbu não foi identificado nenhum dos metabólitos encontrados na semente in natura e este resultado é promissor para a sua utilização como dessalinizador. No seu carvão foi observado apenas ácido L-ascórbico, substância solúvel em água, que além de evitar doenças, é extremamente importante no combate dos radicais livres oriundos de processos oxidativos. Além disso, este material, rico em ácidos graxos, poderá ser mais bem aproveitado, principalmente pela presença do ácido linoléico, considerado benéfico à saúde.
Salvador

Kanadinė jakšūnė nėra gerai ištyrinėtas vaistinis augalas, todėl buvo tikslinga atlikti Lietuvoje išaugintos žaliavos fitocheminius tyrimus augalo veikliųjų junginių sudėčiai nustatyti. Sukaupti duomenys gali būti naudojami iš kanadinės jakšūnės gaminamų preparatų standartizacijai ir naujų, dar nenustatytų farmakologinių poveikių paieškai. Tyrimo objektas ir metodai: kanadinės jakšūnės žaliava ištirta efektyviosios skysčių chromatografijos metodu. Darbo tikslas ir uždaviniai: atlikti kanadinės jakšūnės kaupiamų flavonoidų ir fenilpropanoidų kokybinės ir kiekybinės sudėties tyrimą bei nustatyti, kokiame augalo organe ir kokiu vegetacijos tarpsniu jų kaupiama daugiausiai. Taip pat nustatyti dominuojančius veikliuosius junginius žolėje ir atskiruose organuose bei atlikti palyginamąja ataugusios žaliavos analizę. Ištirti kanadinės jakšūnės žolėje ir lapuose sukauptų flavonoidų suminių kiekių dinamiką skirtingais vegetacijos tarpsniais antraisiais – septintaisiais augimo metais ir pateikti žaliavos rinkimo rekomendacijas bei nustatyti, ar augalo amžius turi įtakos kaupiamų veikliųjų junginių kiekiui. Išvados: Daugiausiai veikliųjų junginių visais vegetacijos tarpsniais susikaupia kanadinės jakšūnės lapuose, išskyrus žydėjimo tarpsnį, kai didžiausias nustatytų veikliųjų junginių kiekis randamas žieduose. Lapuose ir žolėje daugiausiai veikliųjų junginių šeštaisiais augimo metais sukaupta butonizacijos tarpsniu, o stiebuose – vegetacijos pabaigoje. Septintaisiais augimo metais... [toliau žr. visą tekstą]
Showy tick-trefoil is not widely explored medical plant. Therefore it is reasonable to carry out a phytochemical research for the plant which was grown in Lithuania in order to identify its composition of active compounds. Accumulated information of analysis can be useful in standardization of pharmaceutical preparations from showy tick-trefoil, also in researching previously unidentified pharmacological effects. Object and methods: raw material of showy tick-trefoil was examining by methods of high pressure liquid chromatography. Objectives: To perform accumulated flavonoids and fenilpropanoids quality and quantity analysis on showy tick-trefoil and identify in which organ and which vegetation period they are mostly compiled; to identify dominating active compounds in herb and individual organs; to make a comparative analysis of outgrowth raw material. Also to investigate total amounts of compiled flavonoids in showy tick-trefoil herb and leaves in different vegetation periods from second till seventh year of herb growing; to give recommendations for picking raw material. Finally to indicate whether the age of the plant has any influence for quantity of active compounds. Findings: The biggest amount of active compounds in every vegetation period is concentrated in leaves, except blooming period. In the sixth year of growing most of active compounds in leaves and in herb are founded during budding period, in stems – during the end of vegetation. During seventh year of growing... [to full text]

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Embora milhares de espécies vegetais sejam utilizadas na medicina tradicional do mundo, estima-se que apenas 1% são conhecidas por estudos científicos, com valor terapêutico demonstrado. As espécies Polygonum acre e Synadenium carinatum são de fácil cultivo, estando largamente distribuídas no Brasil, e possuindo um alto índice de utilização na medicina popular. Entre os usos populares da espécie Polygonum acre podem ser citados: anti-séptico, antiinflamatório, hipotensor, anti-hemorroidal, diurético, vermicida e anti-diarréico. A espécie Synadenium carinatum é popularmente utilizada para o tratamento de cânceres. No entanto, não existem estudos científicos que comprovem esses efeitos, nem informações sobre a segurança de utilização dessas drogas pelos seres humanos além de haver poucos estudos fitoquímicos destas espécies. Neste trabalho foram realizados os estudos fitoquímico, microbiológico, a busca de atividades biológicas dos extratos dessas plantas e a investigação de aspectos relativos à segurança de utilização. Os testes fitoquímicos indicaram a presença de flavonóides, taninos, saponinas, mono, sesqui e diterpenos, e derivados cinâmicos nas espécies P. acre e S. carinatum, enquanto que apenas a espécie P. acre respondeu positivamente quanto à presença de proantocianidinas condensadas e leucoantocianidinas. Na avaliação da atividade antimicrobiana através do método de difusão em ágar e da técnica de Concentração Inibitória Mínima, a espécie P. acre apresentou atividade antimicrobiana nas concentrações de 300 mg/mL, enquanto a atividade antimicrobiana para os extratos de S. carinatum não foi evidenciada. Nos testes de toxicidade aguda, pode-se observar que na dose de 2 g/kg, o extrato etanólico 70% de P. acre apresentou taxa de mortalidade de 50%, enquanto o extrato etanólico 70% de S. carinatum não apresentou toxicidade...
Although there are thousands of plant species used in traditional medicines in the world, it is estimated that only 1% are known by scientific studies with demonstrated therapeutic value. The species Polygonum acre and Synadenium carinatum are easy to culture, being widely distributed in Brazil, and having a high rate of use in folk medicine. Among the popular uses of the species Polygonum acre may be cited anti-septic, anti-inflammatory, hypotensive, anti-hemorrhoid, diuretic, vermifuge, and anti-diarrheal. The Synadenium carinatum species is popularly used in the treatment against cancer. However, there are no scientific studies that show these effects, no information about the safe use of these drugs by human beings and there are few phytochemical studies of this species. In this work, the phytochemical study and microbiological quality control, the search for biological activities of the extracts of plants and the investigation of aspects related to safe use were done. The phytochemical tests indicated the presence of flavonoids, tannins, saponins, mono, sesqui and diterpenes, and cinnamic derivatives in P. acre and S. carinatum species, while only the species P. acre responded positively about the presence of condensed proanthocyanidins and leucoanthocianidins. In the evaluation of antimicrobial activity by the ágar diffusion method and Minimum Inhibitory Concentration technique, the P. acre species had antimicrobial activity at concentrations of 300 mg / mL, while the antimicrobial activity for the extracts of S. carinatum was not demonstrated. In the tests of acute toxicity the ethanol extract 70% of P. acre at a dose of 2 g/kg showed 50% of mortality while the ethanol extract 70% of S. carinatum has no toxicity. At the dose of 1 g/kg the 70% ethanol extract of P. acre showed 50% of mortality while the latex of S. carinatum caused 100% of death the mice... (Complete abstract click electronic access below)

Ancient medicine is a fast-growing field of research at international level, but since successful research implies both medical (or pharmaceutical) knowledge and the ability to read Latin and Greek, only one classicist in South Africa has published on the topic in co-operation with a medical doctor. Although the professional and scholarly literature on ancient Greek and Roman medicine has proliferated in the last few decades, few studies have appeared that focus on narcotics and analgesics, their effects as observed and recorded by physicians, pharmacologists, and medical botanists, and an evaluation according to modern pharmacology or phytochemistry.1 The study is especially topical in the light of the modern-day search for herbal medicines and the renewed interest in South African ethno botanical traditions (e.g. Afrikaner, Zulu, Swazi) as opposed to chemically prepared narcotics. Phytochemical properties of crude drugs have predictable physiological effects in the living human organism. Therefore, presuming that humans have not changed except incrementally in their physiological chemistry over the short span of two millennia, one can likewise document the effects of narcotics and analgesics in Graeco-Roman pharmacology, provided that the botanical sources and their manufactured forms are carefully identified. A secondary assumption is that ancient plants also carried phytochemical properties generally identical to modern ones, so that the descriptive nomenclatures of modern botany and phytochemistry reveal multiple pharmacological principles and their actions in ancient drugs, as well as their potentially undesirable side effects. 1. One of the exceptions is the 1995 "The Opium Poppy in Hellenistic and Roman Medicine" by Prof. John Scarborough (in Roy Porter and Mikulas Teich, eds., Drugs and Narcotics in History [Cambridge and New York: Cambridge University Press]).
Thesis (Ph.D. (Latin))--North-West University, Potchefstroom Campus, 2009.

Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.

Thesis (PhD(Agric))--Stellenbosch University, 2013.
ENGLISH ABSTRACT: The development and application of science-based tools for determining optimum fruit maturity and postharvest handling protocols to maintain quality and reduce losses during postharvest handling and marketing is essential to maintain the competitiveness of the emerging pomegranate industry in South Africa. Currently, there are no quality standards for the South African pomegranate industry, neither is there a general consensus on the optimal harvest maturity indices for fruit cultivars. These information are important to ensure the delivery of good quality fruit to consumers, particularly for long supply chains. The overall aims of this study were (a) to develop science-based management tools for determining optimum maturity indices and storage performance of pomegranate fruit cultivars grown in South Africa, and (b) to characterise the physico-chemical and pharmacological properties of selected cultivars relevant to postharvest handling and industrial applications. In Section II, seasonal studies on pomegranate (‘Baghwa’ and ‘Ruby’) fruit growth and the evolution of maturity indices during development were conducted. Significant increases in total soluble solids (TSS), sugars (glucose and fructose) and anthocyanin composition, coupled with significant decline in titratable acidity (TA), organic acids and total phenolics (TP) occurred with advancing fruit maturity. Fruit at advanced maturity stages were characterized by intense pigmentation of peel and aril, which coincided with maximum accumulation of anthocyanins. Among all the major maturity indices investigated, TSS, BrimA and anthocyanins did not show significant (p<0.05) seasonal variability, and strong correlations were found among the indices. In combination, these indices accounted for fruit juice sugar content, acidity and colour and could serve as reliable markers to determine optimal maturity for both pomegranate cultivars. The studies in Section III focused on characterization of postharvest quality including nutritional, medicinal and antioxidant properties of fruit parts. Quality attributes of eight commercial cultivars were analysed by cluster analysis, which enabled the cultivars to be separated into two clusters (cluster 1 = ‘Ruby’, ‘Arakta’ and ‘Ganesh’; cluster 2 = ‘Bhagwa’, ‘Acco’ and ‘Herskawitz’) and two ungrouped cultivars (‘Molla de Elche’ and ‘Wonderful’) based on important quality attributes (size, texture, colour, soluble solids, acidity, juiciness and phenolics). Furthermore, pomegranate fruit peel extracts were studied to highlight their potential for value-adding in pharmaceutical and other industrial applications. The results showed that fruit peels of the investigated cultivars possess strong antibacterial, antioxidant and antityrosinase activities, and hence could be exploited as potential sources of natural antimicrobial and antioxidant agents, as well as a potential tyrosinase inhibitor. The research reported in Section IV investigated the effects of harvest maturity and storage conditions on postharvest quality and nutritional value of ‘Bhagwa’ and ‘Ruby’ cultivars. Fruit harvested at commercial maturity were stored at 5±0.3°C, 7±0.5°C and 10±0.4°C with 92±3% RH and at room temperature (20±2.2°C, 65±5.5% RH) for 16 weeks. Fruit physiological responses and quality were affected by storage condition, with the maximum levels of respiration occurring at higher temperature and extended storage duration. Fruit colour and antioxidant capacity varied slightly among storage temperatures, with total soluble solids and titratable acidity decreasing gradually over time at different temperatures. Considering that fruit stored at 5°C and 92% RH had significantly reduced weight loss, low incidence of physiological disorders and best results in maintaining flavour attributes (TSS and TA, TSS:TA ratio), the investigated cultivars may be stored at 5°C and >92% RH for 8 - 12 weeks. In paper 9 (Section IV), the research investigated the relationships between instrumental and sensory measurements of pomegranate fruit at different harvest maturities during storage and shelf life. Mature ‘Bhagwa’ fruit harvested at different times could not be discriminated by sensory attributes assessed by a trained panel. However, TSS (R2 = 0.677) and juice content (R2 = 0.512) were the two most decisive quality attributes at shelf life related to harvest maturity status. For ‘Ruby’, however, a combination of instrumental and sensory attributes appeared to be influential in discriminating mature fruit harvested at different times, with TSS:TA ratio being the most decisive (R2 = 0.654) in distinguishing different fruit harvests, followed by sweet taste (R2 = 0.474) and hue angle (R2 = 0.431). The results showed that to ensure the best post-storage quality of ‘Bhagwa’, the optimum harvest maturity was between 167 - 175 DAFB (H2 and H3) when fruit reached maximum TSS level (>16°Brix; H3) and juice content (>65 mL/100 g aril; H2). However, for ‘Ruby’, this study indicated that the optimum harvest date was at 143 DAFB (H2) when TSS:TA ratio was >55, which coincided with significantly higher sensory rating for sweet taste after shelf life of fruit at H2 than H1 and H3, respectively. The results from this thesis provide new understanding and better insights on fruit characteristics of major pomegranate cultivars grown in South Africa. Overall, the study provides new knowledge on science-based tools for assessing fruit readiness for harvest as well as storage conditions to maintain fruit postharvest quality and reduce losses. It also provides scientific information on phytochemical contents and antioxidant compounds in fruit to promote value-adding of pomegranate as a good raw material with potential applications in health food products and other industrial applications such as pharmaceuticals and cosmetics.
AFRIKAANSE OPSOMMING; Die ontwikkeling en toepassing van wetenskapgegronde instrumente vir die bepaling van optimale vrugrypheid en naoes-hanteringsprotokolle om gedurende die naoes-hantering en -bemarking van vrugte gehalte te behou en verliese te verminder, is noodsaaklik om die mededingendheid van die ontluikende granaatbedryf in Suid-Afrika te verseker. Tans is daar nie enige gehaltestandaarde vir die Suid-Afrikaanse granaatbedryf óf algemene eenstemmigheid oor die optimale oesrypheidsaanwysers vir vrugtekultivars nie. Hierdie inligting is belangrik om die naoes-lewering van uithalervrugte aan verbruikers te verseker, veral vir lang verskaffingskettings. Die oorkoepelende doelwitte van hierdie studie was (a) om wetenskapgegronde bestuursinstrumente te ontwikkel vir die vasstelling van optimale rypheidsaanwysers en bergingsprestasie van granaatkultivars wat in Suid-Afrika verbou word, en (b) om die fisiko-chemiese eienskappe en farmakologiese kenmerke van gekose kultivars te tipeer. In deel II is seisoenale studies oor granaatgroei en die ontwikkeling van rypheidsaanwysers gedurende groei onderneem. Namate vrugte ryp geword het, is beduidende toenames in totale oplosbare vaste stowwe (TSS), suikers (glukose en fruktose) en antosianien-samestelling opgemerk, sowel as ’n beduidende afname in titreerbare suur (TA), organiese suur en totale fenol (TP). Vrugte in gevorderde stadia van rypheid is gekenmerk deur intense pigmentasie van die skil en aril, wat met maksimum opbou van antosianien verband gehou het. Van ál die belangrike rypheidsaanwysers wat ondersoek is, het TSS, BrimA en antosianien onbeduidende (p<0.05) seisoenale veranderlikheid getoon, en is sterk verbande tussen die aanwysers opgemerk. Gesamentlik sou die aanwysers kon rekenskap gee van sapsuikerinhoud, -suurgehalte én -kleur, en sou dit dus as betroubare rypheidsmerkers kon dien om optimale rypheid vir albei granaatkultivars te bepaal. Die studies in deel III het gekonsentreer op die tipering van die naoes-kenmerke, onder meer die voedings-, medisinale en antioksidant-kenmerke van vrugtedele. Kenmerke van agt kommersiële kultivars is deur middel van groepsontleding bestudeer, waarvolgens die kultivars op grond van belangrike kenmerke (grootte, tekstuur, kleur, oplosbare vaste stowwe, suurgehalte, sappigheid en fenol) in twee groepe (groep 1 = ‘Ruby’, ‘Arakta’ en ‘Ganesh’; groep 2 = ‘Bhagwa’, ‘Acco’ en ‘Herskawitz’) en twee niegegroepeerde kultivars (‘Molla de Elche’ en ‘Wonderful’) ingedeel is. Ten einde die toegevoegde waarde van granaatskille vir farmaseutiese en kosmetiese doeleindes te bevorder, is skilekstrakte ook bestudeer. Die resultate toon dat die vrugteskille van die bestudeerde kultivars oor sterk antibakteriese, antioksidant- en antitirosinase- eienskappe beskik. Daarom kan die skil van die granaatkultivars as moontlike bron van natuurlike antimikrobiese en antioksidant-agense sowel as ’n moontlike tirosinase-inhibitor ontgin word. Die navorsing in deel IV het ondersoek ingestel na die uitwerking van oesrypheid en bergingsomstandighede op die naoes-gehalte en -voedingswaarde van die kultivars ‘Bhagwa’ en ‘Ruby’. Vrugte wat op kommersiële rypheid geoes is, is vir 16 weke by 5±0.3 °C, 7±0.5 °C en 10±0.4 °C met 92±3% RH, sowel as by kamertemperatuur (20±2.2 °C, 65±5.5% RH) geberg. Die bergingsomstandighede het die fisiologiese reaksies en gehalte van die vrugte beïnvloed: Maksimum vlakke van respirasie het teen hoër temperature en met verlengde berging voorgekom. Die kleur en antioksidantvermoë van die vrugte het effens tussen bergingstemperature verskil, en totale oplosbare vaste stowwe en titreerbare suur het mettertyd geleidelik by verskillende temperature afgeneem. Gedagtig daaraan dat die vrugte wat teen 5 °C en 92% RH geberg is beduidend minder gewigsverlies, ’n lae voorkoms van fisiologiese afwykings en die beste resultate in blywende geurkenmerke (TSS en TA, TSS:TA-verhouding) getoon het, kan die bestudeerde kultivars vir 8 tot 12 weke teen 5 °C en >92% RH geberg word (navorsingstuk 8). In navorsingstuk 9 (deel IV) is daar ondersoek ingestel na die verhouding tussen instrument- en sintuiglike metings van granate in verskillende stadia van oesrypheid gedurende berging en raklewe. Geen verskil in sintuiglike kenmerke kon bespeur word by ryp ‘Bhagwa’- vrugte wat op verskillende tye geoes is nie. Tog was TSS (R2 = 0.677) en sapinhoud (R2 = 0.512) die twee bepalendste gehaltekenmerke wat betref oesrypheidstatus gedurende raklewe. By ‘Ruby’ kon ’n kombinasie van instrument- en sintuiglike kenmerke egter wél tussen stadia van oesrypheid onderskei, met die TSS:TA-verhouding die bepalendste (R2 = 0.654) in die onderskeid tussen verskillende vrugteoeste, gevolg deur ’n soet smaak (R2 = 0.474) en skakeringshoek (R2 = 0.431). Die resultate toon dat die beste nabergingsgehalte vir ‘Bhagwa’ verkry word by ’n optimale oesrypheid van 167–175 DAFB (H2 en H3), wanneer vrugte die maksimum TSS-vlak (>16°Brix; H3) en sapinhoud (>65 mL/100 g aril; H2) bereik het. Vir ‘Ruby’ dui hierdie studie op ’n optimale oesdatum van 143 DAFB (H2) met ’n TSS:TA-verhouding van >55, wat verband gehou het met ’n beduidend hoër telling vir soet smaak by H2 eerder as by H1 en H3 ná raklewe. Die resultate van hierdie tesis bied ’n beter begrip van, en insig in, die vrugtekenmerke van granaatkultivars wat in Suid-Afrika verbou word. Oor die algemeen bied die studie wetenskaplike inligting om moontlik die toegevoegde waarde van granate as ’n goeie bron van minerale elemente sowel as farmaseutiese, kosmetiese en antioksidant-verbindings te bevorder. Dit bied ook kennis oor die ontwikkeling van wetenskapgegronde instrumente vir die vasstelling van optimale vrugrypheid en naoes-hanteringsprotokolle om gedurende die naoes-hantering en -bemarking van granate vruggehalte te behou en verliese te verminder.

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CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Leishmaniasis is a world public health problem wide considered by the World Health Organization as one of five infectious endemic diseases of major relevancy. The search for leishmanicids agents with fewer side effects is one of the greatest challenges in modern research, and is increasingly related to the uses of medicinal plants and other natural products. The species Caesalpinia ferrea Martius widely distributed in Brazil it is popularly used for many therapeutic purposes, including the treatment of various infections like those caused by leishmaniasis, making it a promising tests of biological activity against leishmania. The objective of this study was to evaluate by bioassays the activity of hexane and methanol extracts against protozoa, analyzing semi-purified fractions obtained from the most active. The fruits of C. ferrea were collected at the National Institute of Amazon Research, V8’s Campus in the City of Manaus - AM and processed in the departments of Natural Products and leishmaniasis and Chagas Diseases at INPA. The studies were performed with the whole fruit and its separate parts, coconut shell and seed. Hexane e methanol crude extracts were prepared from the fruits, coconut shell and seeds, which were tested to evaluate the biological activity in vitro against promastigotes of the stationary phase of the protozoan Leishmania (Viannia) guyanensis. The extracts were analyzed by thin layer chromatography Comparison (CCDC) and Nuclear Magnetic Resonance (NMR) to determine the chromatographic profile. All biological assays were carried out experimentally in vitro in microplates containing 106/mL promastigotes to axenic cultures of L. (V.) guyanensis evaluating the inhibitory activity and stimulating parasite growth. The most active crude extract was submitted to CCDC and fractionation by partition líquid-líquid resulting in semi-purified fractions that were evaluated for their chemical composition. The results showed that the epicarp methanolic extract of C. ferrea, showed anti-leishmanial activity for L. (V.) guyanensis in the concentration of 20 mg/mL/24h. However, contrary to expectations, there was a stimulatory effect on L. (V.) guyanensis by the hexanic extract obtained from the fruit seed of C. ferrea. Chemically analyzing the coconut shell and fruits methanolics extracts of, which showed anti-leishmanial activity, we verified the presence of substances such as chlorophyll, flavonoids, xanthones, tannins, triterpenes and saponins as major constituents, and shows antioxidant activity. The results showed that the methanolics extracts of coconut shell and fruitsof C. ferrea can be considered as an promising alternative in the treatment of cutaneous leishmaniasis and should be undertaken pharmacological and toxicological tests in vivo of extract, making other fractioning in order to isolate the substances responsible for the anti-leishmanial activity.
A leishmaniose é um problema de saúde pública mundial considerada pela Organização Mundial da Saúde como uma das cinco doenças infecto-parasitárias endêmicas de maior relevância. A busca por agentes leishmanicidas com poucos efeitos colaterais é um dos maiores desafios na pesquisa moderna, e cada vez mais está relacionada aos usos de plantas medicinais e outros produtos naturais. A espécie Caesalpinia ferrea Martius largamente distribuída no Brasil, é usada popularmente para muitos fins terapêuticos, inclusive no tratamento de diversas infecções como as causadas pela Leishmaniose, tornando-a promissora para a realização de ensaios de atividade biológica contra a leishmania. O objetivo deste trabalho foi o de avaliar, através de bioensaios, a atividade de extratos hexânicos e metanólicos contra protozoários, analisando também frações semi-purificadas obtidas do extrato de melhor atividade leishmanicida. Os frutos de C. ferrea foram coletados no Instituto Nacional de Pesquisas da Amazônia, Campus do V8 na Cidade de Manaus-AM, e processados nos departamentos de Produtos Naturais e de Leishmaniose e Doença de Chagas no INPA. Os estudos foram realizados com os frutos inteiros e suas partes separadas, epicarpos e sementes. Foram preparados extratos brutos hexânicos, e metanólicos a partir dos frutos inteiros e também das partes separadas, epicarpos e sementes, os quais foram submetidos aos ensaios para avaliação da atividade biológica in vitro contra formas promastigotas da fase estacionária de protozoários da espécie Leishmania (Viannia) guyanensis. Os extratos brutos foram analisados através de Cromatografia em Camada Delgada Comparativa (CCDC) e Ressonância Magnética Nuclear (RMN) para determinação do perfil cromatográfico. Todos os ensaios biológicos foram realizados experimentalmente in vitro, em microplacas contendo 106/mL formas promastigotas de culturas axênicas de L. (V.) guyanensis (MHOM/BR/75/IM 4147) avaliando-se a atividade inibitória ou ativadora de crescimento parasitário. O extrato bruto de melhor atividade biológica foi submetido à CCDC e fracionamento por partição líquido-líquido obtendo-se frações semi-purificadas que foram avaliadas quanto a sua composição química. Os resultados mostraram que o extrato metanólico do epicarpo de C. ferrea, apresentou atividade leishmanicida para L. (V.) guyanensis na concentração de 20 mg/mL/24h. No entanto, ao contrário das expectativas, houve um efeito estimulante em L. (V.) guyanensis pelo extrato hexânico obtido das sementes de frutos de C. ferrea. Analisando quimicamente o extrato metanólico de frutos e epicarpos, os quais demonstraram atividade leishmanicida, verificou-se a presença de substâncias como a clorofila, flavonóides, xantonas, taninos, triterpenos e saponinas como constituintes majoritários, além de apresentar atividade antioxidante. Os resultados obtidos demonstraram que os extratos metanólicos de frutos e epicarpos de C. ferrea podem ser considerados como importantes alternativas no tratamento das leishmanioses, devendo ser realizado testes farmacológicos e toxicológicos in vivo desses extratos, realizando outros fracionamentos a fim de se confirmar alguma substância de melhor atividade leishmanicida.

Les plantes médicinales constituent une source inexhaustible de composés (des produits naturels - PN) utilisé en médecine pour la prévention et le traitement de diverses maladies. L'introduction de nouvelles technologies et méthodes dans le domaine de la chimie des produits naturels a permis le développement de méthodes ‘high throughput’ pour la détermination de la composition chimique des extraits de plantes, l'évaluation de leurs propriétés et l'exploration de leur potentiel en tant que candidats médicaments. Dernièrement, la métabolomique, une approche intégrée incorporant les avantages des technologies d'analyse moderne et la puissance de la bioinformatique s’est révélé un outil efficace dans la biologie des systèmes. En particulier, l'application de la métabolomique pour la découverte de nouveaux composés bioactifs constitue un domaine émergent dans la chimie des produits naturels. Dans ce contexte, le genre Acronychia de la famille des Rutaceae a été choisi sur la base de son usage en médecine traditionnelle pour ses propriétés antimicrobienne, antipyrétique, antispasmodique et anti-inflammatoire. Nombre de méthodes chromatographiques modernes, spectrométriques et spectroscopiques sont utilisées pour l'exploration de leur contenu en métabolites suivant trois axes principaux constituant les trois chapitres de cette thèse. En bref, le premier chapitre décrit l’étude phytochimique d’Acronychia pedunculata, l’identification des métabolites secondaires contenus dans cette espèce et l'évaluation de leurs propriétés biologiques. Le deuxième chapitre vise au développement de méthodes analytiques pour l'identification des dimères d’acétophénones (marqueurs chimiotaxonomiques du genre) et aux stratégies utilisées pour la déréplication de ces différents extraits et la caractérisation chimique des composés par UHPLC-HRMSn. Le troisième chapitre se concentre sur l'application de méthodologies métabolomique (RMN et LC-MS) pour l'analyse comparative (entre les différentes espèces, origines, organes), pour des études chimiotaxonomiques (entre les espèces) et pour la corrélation des composés contenus avec une activité pharmacologique
Medicinal plants constitute an unfailing source of compounds (natural products – NPs) utilised in medicine for the prevention and treatment of various deceases. The introduction of new technologies and methods in the field of natural products chemistry enabled the development of high throughput methodologies for the chemical composition determination of plant extracts, evaluation of their properties and the exploration of their potentials as drug candidates. Lately, metabolomics, an integrated approach incorporating the advantages of modern analytical technologies and the power of bioinformatics has been proven an efficient tool in systems biology. In particular, the application of metabolomics for the discovery of new bioactive compounds constitutes an emerging field in natural products chemistry. In this context, Acronychia genus of Rutaceae family was selected based on its well-known traditional use as antimicrobial, antipyretic, antispasmodic and anti-inflammatory therapeutic agent. Modern chromatographic, spectrometric and spectroscopic methods were utilised for the exploration of their metabolite content following three basic axes constituting the three chapters of this thesis. Briefly, the first chapter describes the phytochemical investigation of Acronychia pedunculata, the identification of secondary metabolites contained in this species and evaluation of their biological properties. The second chapter refers to the development of analytical methods for the identification of acetophenones (chemotaxonomic markers of the genus) and to the dereplication strategies for the chemical characterisation of extracts by UHPLC-HRMSn. The third chapter focuses on the application of metabolomic methodologies (LC-MS & NMR) for comparative analysis (between different species, origins, organs), chemotaxonomic studies (between species) and compound-activity correlations

Magister Scientiae (Biodiversity and Conservation Biology)
South Africa is recognised worldwide for its rich diversity of plants, many of which have been used in ethno-medicine. However, the use of wetland plant species in ethno-medicine required further investigations. This research is aimed at investigating three wetland medicinal, plant species, Centella asiatica, Cyperus longus and Typha capensis based on their geographical, seasonal, mineral nutrient (Fe, Mn, Ca, Mg, K and Na) and secondary metabolite characteristics. Samples of each species were collected from Grabouw, Kelderhoff, Kenilworth, Pringle Bay, University of the Western Cape (UWC) and Worcester within the Western Cape Province of South Africa. Specimen and soil collections were carried out during autumn, spring, summer and winter of 2014. Both plant and soil samples were acid digested and mineral nutrient concentrations in the samples were analysed using an atomic absorption spectrometer (AAS). Secondary metabolites were determined using analytical TLC on normal phase Merck-Silva gel coated aluminium plates as well as by using HPLC separation from crude extracts of C. asiatica, C. longus and T. capensis using LC-MS hardware from Agilent. The elemental analysis of soil samples showed that Ca, K, Mg, Mn and Zn concentrations were predominantly low. Soil mineral concentrations increased progressively from inland (Worcester) towards the coastland in the south (Pringle Bay). Calcium and sodium concentrations, in particular, were higher in soil samples obtained from Grabouw (inland south) and decreased northward towards Worcester. Comparatively, plant mineral concentrations were generally higher than soil concentrations. The high concentration of some of these essential elements, in selected plants is an indication that these plant species could be a good source of essential elements. High concentrations of phytochemicals were found in Centella asiatica during winter, while Cyperus longus and Typha capensis exhibited high concentrations during autumn indicating variation in respect of season. Consequently, harvesting of the studied plants should be done at the season with a relatively high phytochemical concentration. Studies are needed to investigate the extent of pesticide or herbicide contamination in wetland plants to protect the health of users. The LC-MS analyses of the three study species showed that seasonal variation affects metabolite constituents and moreover that these metabolite constituents differ from one locality to another. The seasonal variation of the elements in the studied medicinal plants justified the importance of harvesting seasons in the optimal utilization of the studied plants for medicinal purpose. s, for C. asiatica, anti-bacterial treatments for C. longus and fertility enhancement and birth control for T. capensis.

Syzygium cordatum is a medicinal plant indigenous to South Africa and Mozambique, commonly used to treat stomach aches, diabetes, respiratory problems and tuberculosis. In spite of the folklore use, adequate scientific data to credit its widespread traditional use is lacking. The objectives of this study were: to evaluate and validate scientifically the successful therapeutic claims by traditional medicine practitioners that Syzygium cordatum is effective in treating diarrhoea and diabetes
to determine the effects of the plant extract on gastrointestinal transit of a charcoal meal in mice
to determine the effects on castor oilinduced intestinal fluid accumulation
to determine the safety profile of the plant by carrying out acute toxicology study and to carry out preliminary screening of the active compounds present in the plant using standard phytochemical analytical procedures. The aqueous leaf extract of Syzygium cordatum (3.125 -50mg/kg, p.o) significantly reduced the faecal output caused by castor oil (0.7ml). All the doses used, reduced faecal output from 100% produced by castor oil to between 40 and 61%. S.cordatum (6.25 &ndash
50mg/kg, p.o) significantly and in a dose dependent manner, delayed the onset of castor oil-induced diarrhoea.

Background. As a result of the AIDS pandemic, many people areimmuno compromised andopportunistic fungal infections (OFIs) such as candidiasis are common. Despite the widespread use of medicinal plants in South Africa, there is a dearth of knowledge regarding the use of such plants in the management of these infections. This study evaluates three South African medicinal plants (Arctotis arctotoides, Pittosporum viridiflorum, and Gasteria bicolor) traditionally used in the treatment of OFIs in HIV/AIDS patients, in the Eastern Cape Province, South Africa. Materials and methods. A six-stage process of documentation, evaluation and analysis of results was conducted: (1) Selection of medicinal plants most frequently used in the treatment of OFIs through ethnomedical studies and the survey of specialised literature; (2) Collection and preparation of the extract of each plant; (3) Antifungal evaluation of the crude plant extracts. (4) Phytochemical and antioxidant evaluation of the active crude plant extracts; (5) Cytotoxicity evaluation of the bioactive extracts using the Chang liver cell line, and (6) Statistical analysis of the results. Ethnobotanical information was obtained through interviews with traditional healers and AIDS patients with the aid of semi-structured questionnaires, direct observations and by reviewing studies reported in the literature. Following the approval from the University of Fort Hare‘s Ethics Committee, 101 HIV/AIDS patients were recruited through convenience sampling into an anonymous cross-sectional questionnaire study. The agar diffusion and micro-dilution methods were used to determine the antifungal activities of the hexane, acetone and aqueous extracts of A. arctotoides, G. bicolor and P. viridiflorum against 10 opportunistic fungi.

L'objectif de cette thèse est d'évaluer les modifications du métabolisme secondaire des plantes contaminées aux éléments trace métalliques (ETM) et leurs conséquences sur les communautés bactériennes rhizosphériques associées incluant des bactéries présentant des phénotypes de MultiDrug Résistance (MDR). Nous nous sommes focalisés sur deux contextes de sols exposés aux métaux : la phytoremédiation de sites miniers au Vietnam et la reconversion de sols agricoles contaminés par la re-déposition atmosphérique d'activités métallurgiques en France. Nos résultats ont mis en évidence que la contamination par différents types de métaux (dont Cu et Pb principalement) a conduit à une altération de la production des métabolites secondaires des racines, tiges et feuilles de la plante hyperaccumulatrice Pteris vittata et que concernant les racines des tendances similaires dans les changements métaboliques ont pu être observés dans un autre type de contexte de pollution (Zn et Pb plus particulièrement). De même, les profils métaboliques des parties souterraines (racines et rhizomes) de Miscanthus x giganteus ont été modifiés par les concentrations en Pb, Cd et Zn des sols agricoles. Pour les deux plantes examinées des dérivés de l'acide chlorogénique ont été retrouvés en proportions augmentées dans les racines malgré des contextes de nature des sols et de pollutions métalliques très contrastés. Cependant, les dérivés de tanin catéchiques sont spécifiquement trouvés en proportions plus élevées dans les racines de P. vittata sous pression métallique. Ces polyphénols sont connus pour leur capacité à piéger les radicaux libres et leur pouvoir antioxydant et pourraient donc être impliqués dans l'adaptation de ces plantes au stress métallique en contribuant à limiter le stress oxydatif généré par les ETM. Au niveau des parties aériennes, nous n'avons étudié que le changement pour P. vittata et avons mis en évidence une proportion plus élevée de dérivés flavonoïdiques pour les plantes contaminées. Nos résultats de métagénomique nous permettent de conclure également sur un effet des ETM sur la diversité et la richesse spécifique des communautés bactériennes des sols étudiés : une forte contamination en Cu (10 fois la limite autorisée) a diminué la diversité et la richesse bactérienne, alors que pour des niveaux en ETM plus modérés incluant Cu, Pb et Zn, la diversité des communautés bactériennes rhizosphériques semble plus influencée par la plante ou la saison plutôt que par l'effet des ETM. Cet effet sur la composition bactérienne de la rhizosphère de P. vittata se traduit par un enrichissement de certains genres connus comme pathogènes opportunistes de l'homme, notamment Ralstonia, Acinetobacter, Burkholderia et Mycobacterium. En outre, le genre Cupriavidus, connu comme très résistant aux ETM est le seul genre spécifiquement associé à P. vittata qui ait été augmenté au sein de la communauté rhizosphérique pour les deux sites miniers étudiés par rapport aux sols rhizosphériques non pollués. Ce genre pourrait donc être impliqué dans le processus d'adaptation de cette plante au stress métallique. Quant aux communautés rhizosphériques de Miscanthus x giganteus, la sélection de Stenotrophomonas et Pseudomonas dans les sols agricoles contaminés a été observée. Dans le cadre de cette thèse nous avons également mis au point une méthode rapide pour tester l'impact de métabolites végétaux sur des souches pathogènes d'origine clinique et environnementale et également évaluer leur activité inhibitrice de pompes à efflux (IPE) de la famille des RND. Nos données ont ainsi permis de mettre en évidence des activités intéressantes et comparables à celle de l'inhibiteur de pompe à efflux PAßN pour des composés testés qui étaient extraits des racines de Fallopia x bohemica ou des dérivés de ces derniers
The objective of this thesis is to evaluate the modification of plant secondary metabolism production contaminated with metallic trace elements (MTE) and its consequences on the associated rhizospheric bacterial communities including bacteria presenting MultiDrug Resistant (MDR) phenotypes. We have focused on two contexts of metals exposure: the phytoremediation of mining sites in Vietnam and the reconversion of agricultural soils contaminated by the atmospheric re-deposition of metallurgical activities in France. Our results highlighted that contamination by different types of metals (mainly Cu and Pb) has led to an alteration in the production of secondary metabolites in the roots, stems and leaves of the hyper-accumulating Pteris vittata and for roots, a similar trend in the metabolic changes could be observed in another type of pollution context (Zn and Pb more particularly). Similarly, the metabolic profiles of the underground parts (roots and rhizomes) of Miscanthus x giganteus were modified by the concentrations of Pb, Cd and Zn in agricultural soils. For the two plants examined chlorogenic acid derivatives have been found in increased proportions in the roots despite soil type and pollution context were highly contrasted. However, catechic tannin derivatives are specifically found in higher proportions in the roots of P. vittata under metal pressure. These polyphenols are known for their ability to scavenge free radicals and their antioxidant properties and thus could be involved in the adaptation of these plants to metallic stress by helping to limit the oxidative stress generated by MTE. At the level of the aerial parts, we studied only the change for P. vittata and evidenced higher proportions of flavonoid derivatives for contaminated plants. Our metagenomic results allow us to conclude also on the effect of MTE on the diversity and the specific richness of the bacterial communities of the studied soils: a high contamination of Cu (10 times the allowed limit) decreased dramatically bacterial richness and diversity, while for more moderate MTE levels including Cu Pb and Zn, the diversity of rhizosphere bacterial communities was more explained by plant or season effect rather than an effect of MTE. This effect on P.vittata rhizosphere bacterial composition is reflected by an enrichment in genera known as opportunistic human pathogens, including Ralstonia, Acinetobacter, Burkholderia and Mycobacterium. In addition, Cupriavidus, known as a highly resistant genus, is the only P. vittata specifically associated genus found in increased proportions at both mining sites compared to non-contaminated rhizosphere soils. This genus could then be involved in the adaptation process of this plant with metal stress. As for the rhizospheric communities of Miscanthus x giganteus, the selection of Stenotrophomonas and Pseudomonas in agricultural soils contaminated with MTE was observed. As a part of this thesis, we have also developed a rapid method for testing the impact of plant metabolites on pathogenic strains of clinical and environmental origin and their efflux pump inhibition (EPI) activity of RND family. Our data thus showed interesting and notable EPI activities comparable to that of the efflux pump inhibitor PAßN for tested compounds issued from Fallopia x bohemica roots or for their derivatives

La galanthamine est un alcaloïde isoquinoléique, utilisé dans le monde entier pour le traitement palliatif de la maladie d’Alzheimer en raison de son pouvoir inhibiteur de l’acétylcholinestérase. Cet alcaloïde est extrait à partir de bulbes d’Amaryllidaceae, Leucojum aestivum, Galanthus nivalis, et Narcissus sp. ou obtenu par synthèse chimique. La difficulté principale de cette dernière réside dans le respect de la configuration des centres d'asymétrie. La culture de tissus in vitro pourrait constituer une alternative intéressante pour obtenir ce composé à haute valeur ajoutée. Le premier objectif de ce projet, vise à améliorer l’accumulation de cet alcaloïde par les biais des biotechnologies. Le second objectif est de rechercher par criblage phytochimique (HPLC, LCMS, GCMS, et HPTLC-MS) de bulbes in vitro et in vivo d’Amaryllidaceae, de nouveaux alcaloïdes biologiquement actifs. Le troisième objectif porte sur l’étude de la voie de biosynthèse en vue de réaliser une synthèse biomimétique de la galanthamine. Nous avons établi des cultures in vitro de 3 espèces d’Amaryllidaceae. La variation des paramètres exogènes a conduit à une accumulation accrue d’alcaloïdes (0,02 à 0,2 % MS). Le criblage phytochimique a conduit à l’identification d’alcaloïdes nouveaux issus des cultures in vitro, n’existant pas in vivo, et présentant un puissant pouvoir inhibiteur de l’acétylcholinestérase (40 à 80 % Inh). L’ajout, de la 4’-Ométhyl-d3-norbelladine aux cultures in vitro a conduit à sa métabolisation en trois types d’alcaloïdes deutérés. Une stimulation de la synthèse et du relargage de la galanthamine native (0,15 % MS et 0,16 % Milieu) a été observée en présence du précurseur deutéré
Galanthamine is an Amaryllidaceae alkaloid used worldwide for the symptomatic treatment of Alzheimer’s disease because of his capacity to inhibit the acetylcholinesterase enzyme. There are two galanthamine sources for medical applications. One is the total synthesis, a complicated process because galanthamine has three asymmetric carbons, requiring stereochemically controlled synthesis. Galanthamine is also extracted from bulbs of some Amaryllidaceae such as Leucojum aestivum, Galanthus nivalis, and Narcissus sp.. The first aim of this work is to improve the accumulation of this alkaloid using biotechnologies. The second aim consists on the phytochemical screening (HPLC, LCMS, GCMS, et HPTLCMS) of in vivo and in vitro Amaryllidaceae bulbs, in order to identify new alkaloids with important pharmacological activities. Finally, the third aim concerns the study of the biosynthesis pathway in order to establish a biomimetic synthesis of galanthamine. Therefore, we established in vitro cultures of three Amaryllidaceae species. The variation of exogenousparameters led to the obtainment of high galanthamine accumulation (0.02 to 0.2 % DW). The phytochemical screening showed new alkaloids in extracts of in vitro cultures, which did not exist in in vivo extracts, and possessing high acetylcholinesterase activity (40 to 80 % Inh). The 4’-O-methyl-d3-norbelladine is incorporated into three different groups ofAmaryllidaceae alkaloids. The addition of the labelled precursor to shoot cultures stimulated the synthesis of native galanthamine (0.15 % DW and 0.16 % Culture medium)

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FUNDAÇÃO DE AMPARO À PESQUISA E AO DESENVOLVIMENTO CIENTIFICO E TECNOLÓGICO DO MARANHÃO
Phytotherapy, for being traditional practice of health recognized in several scientific studies and for World Health Organization, has become as an pharmacological alternative well accepted and accessible to the world s populations. As a result of this therapeutic practice consolidation, several plant species and their products are commercialized at fairs and public markets. The specie Schinus terebinthifolius Raddi, popularly known as aroeira, is among the plants widely consumed, due to its properties anti-inflammatory, healing and antibacterial. Thus, this study aims to evaluate the quality of commercial samples of aroeira, acquired in public markets in the city of São Luís - MA. For the study were used aroeira's barks acquired in twelve public markets, as well as barks obtained of a specimen previously identified in herbarium, used as reference sample of the specie Schinus terebinthifolius Raddi. Was performed analysis morphological (macroscopic) and anatomical (microscopic) of the cross sections and powders obtained from the barks, phytochemicals tests, evaluation of moisture content and antibacterial activity by the methods of broth dilution and agar diffusion. After macroscopic analysis, six commercial samples showed morphological aspects different from reference sample. With regard to microscopic analysis was demonstrated the authenticity of five commercial samples, by comparison with the plant reference sample and information in the literature. In Phytochemical analysis, the reference sample showed the major constituents characteristic of the species, however, the commercial samples analysis, differences were found related, mainly, the analysis of tannins and saponins. The differences observed in this evaluation may reflect the influence of intrinsic and extrinsic factors on the chemical composition of plant species. The determination of moisture in the commercial samples analyzed showed that five had moisture content above the maximum limit allowed for barks, being, therefore, reproved in thismcriterion. In the evaluation of antibacterial activity, all samples showed some degree of activity for Staphylococcus aureus ATCC 25923, however, only six were also active against Escherichia coli ATCC 25922, four of those considered authentic in the microscopic analysis. The verification of poor quality of herbal drugs marketed for therapeutic purposes demonstrates the need for development of inspection programs, surveillance and quality control of these drugs, aiming, thus, consumer safety.
A fitoterapia, por ser prática tradicional de saúde reconhecida em diversos estudos científicos e pela Organização Mundial da Saúde, tem se constituído como uma alternativa medicamentosa bem aceita e acessível aos povos do mundo. Como consequência da consolidação desta prática terapêutica, várias espécies vegetais e seus produtos são comercializados em feiras e mercados públicos. A espécie Schinus terebinthifolius Raddi, popularmente conhecida como aroeira, está entre as plantas de grande consumo devido à suas propriedades anti-inflamatória, cicatrizante e antibacteriana. Assim, o objetivo deste estudo foi avaliar a qualidade de amostras comerciais de aroeira, adquiridas em mercados públicos da cidade de São Luís - MA. Para o estudo foram utilizadas cascas de aroeira adquiridas em doze mercados públicos, bem como cascas obtidas de um exemplar previamente identificado em herbário, utilizadas como amostra referência da espécie S. terebinthifolius Raddi. Realizou-se análise morfológica (macroscópica) e anatômica (microscópica) das secções transversais e dos pós obtidos das cascas, testes fitoquímicos, avaliação do teor de umidade e da atividade antibacteriana pelos métodos de diluição em caldo e difusão em ágar. Após análise macroscópica, seis amostras comerciais apresentaram aspectos morfológicos diferentes da amostra referência. No que se refere à análise microscópica, ficou demonstrado a autenticidade de cinco amostras comerciais, através de comparação com a amostra referência e de informações contidas em literatura. Na análise fitoquímica, a amostra referência apresentou os principais constituintes característicos da espécie, no entanto, na análise das amostras comerciais, constatou-se diferenças relacionadas, principalmente, a análise de taninos e saponinas. As diferenças observadas nesta avaliação podem ser reflexo da influência dos fatores intrínsecos e extrínsecos sobre a composição química de espécies vegetais. A determinação de umidade nas amostras comerciais analisadas revelou que cinco apresentavam teor de umidade acima do limite máximo permitido para cascas, ficando, portanto, reprovadas neste critério. Na avaliação da atividade antibacteriana, todas as amostras revelaram algum grau de atividade para Staphylococcus aureus ATCC 25923, no entanto, apenas seis foram ativas também contra Escherichia coli ATCC 25922, quatro dessas consideradas autênticas na análise morfoanatômica. A comprovação da má qualidade de drogas vegetais comercializadas com fins terapêuticos demonstra a necessidade de desenvolvimento de programas de fiscalização, vigilância e controle de qualidade dessas drogas, objetivando, assim, a segurança do consumidor.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
As pimentas do gênero Capsicum spp. fazem parte da biodiversidade e da riqueza cultural brasileira, são cultivadas em todo o território nacional possuindo uma enorme variedade de cores, sabores, tamanho e pungência. A Embrapa Clima Temperado – Pelotas, com o intuito de conservar e caracterizar a variabilidade genética de pimentas do gênero Capsicum, mantém um banco ativo de germoplasma de Capsicum (BAG). As pimentas são utilizadas in natura ou processadas. Portanto muitos destes genótipos necessitam de avaliação quanto aos aspectos de qualidade e potencial para utilização em produtos alimentícios. Durante o processamento, pode ocorrer redução no teor de compostos bioativos, pela sua instabilidade às condições tecnológicas de processamento, o que consequentemente pode reduzir a capacidade antioxidante, além de interferir também nas características sensoriais. Com base nisto o presente trabalho teve como objetivo caracterizar acessos de pimenta Capsicum spp. para fins tecnológicos e avaliar a estabilidade de compostos potencialmente bioativos. O teor de compostos fenólicos de pimentas in natura variou de 52,45 mg.100g -¹ (P259) a 21,20 mg.100g -¹ (P27) equivalentes de ácido gálico. O teor de carotenoides totais variou de 0,64 mg.100g -¹ (P259) a 40,26 mg.100g -¹ (P115) equivalentes de β-caroteno, o teor de ácido L-ascórbico de 158,38 mg.100g -¹ (P22) a 18,86 mg.100g -¹ (P115) e o teor de capsaicinoides totais de 0,024 mg.100g -¹ (3,93 SHU) no acesso P27 a 1734 mg.100g -¹ (279.174,27 SHU) para o acesso P247. Em relação aos produtos observaram-se perdas e incrementos nos teores de compostos bioativos, os quais foram influenciados pelo tipo de processamento e pelas características iniciais do fruto in natura (principalmente para geleia e pirulito , quando comparado com teor do fruto in natura). A redução do teor de vitamina C (ácido L ascórbico), chegou a 81,62% para a conserva da pimenta P259, e o teor de compostos fenólicos totais em conserva de pimenta P115 foi 5 vezes menor quando comparado ao fruto in natura. O processamento de pimenta conserva, desidratada e em pasta promoveu o incremento no teor de carotenoides totais. A análise sensorial dos produtos revelou que os consumidores de pimenta da região de Pelotas/RS preferem produtos com pungência não muito elevada e de coloração vermelha.
Capsicum spp., part of the Brazilian biodiversity and cultural richness, are grown all over the country having a huge variety of colors, flavors, size and pungency. Embrapa Temperate Climate - Pelotas, in order to conserve and characterize the variability of Capsicum, maintains an active germplasm bank of Capsicum (BAG). The peppers are used fresh or processed. Therefore many of these genotypes require evaluation of quality attributes and potential for use in food products. During processing, a reduction in the content of bioactive compounds, due to their instability to technological processing conditions, can consequently reduce their antioxidant capacity, and also interfere with sensory characteristics. The present study aimed to characterize pepper Capsicum spp. accessions for technological purposes and to evaluate the stability of potentially bioactive compounds. The content of phenolic compounds in fresh pepper fruit ranged from 52.45 mg.100g -¹ (P259) to 21.20 mg.100g -¹ (P27) of gallic acid equivalents. The total carotenoid content ranged from 0.64 mg.100g -¹ (P259) to 40.26 mg 100g -¹ (P115) of β-carotene, the content of L-ascorbic acid from 158.38 mg 100g -¹ (P22) 18.86 mg.100g -¹ (P115) and total capsaicinoids content from 0.024 mg.100g -¹ (3,93 SHU) in accession P27 to 1734 mg.100g -¹ (279.174,27 SHU) for the P247 accession. For pepper products, losses and increments in the content of bioactive compounds were observed influenced by the type of processing and the initial characteristics of the fresh fruit (mainly for jelly and lollipops when compared to the content of the fresh fruit). Vitamin C (L-ascorbic acid) reduction was 81.62% for the pickled pepper P259, and total phenolics content of pickled pepper P115 was 5 times lower when compared to the fresh fruit. The processing of pickled, dehydrated and pepper sauce promoted the increase in the levels of total carotenoids. The sensory analysis of the products revealed that consumers of pepper in Pelotas/RS prefer products medium pungency and red color.

Ethanol extracts of “jenipapo” (Genipa americana Linnaeus), “umbu" (Spondia tuberosa Arruda), “siriguela” (Spondia purpurea Linnaeus) and “mangaba” (Hancornia speciosa Gomes) were prepared from separate pulp, seeds and peel; except mangaba which are used pulp and peel. The investigation of antioxidant capacity of the ethanol extracts were carried out by the methods of determining the content of ascorbic acid and total phenolics; scavenging 2, 2 -diphenyl-1-picrilhydrazyl and 2, 2’- azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical, antioxidant capacity of reduction of iron and copper and lipid peroxidation using a biomimetic membrane system, and measurement enzymatic of catalase and superoxide dismutase. It was also analyzed the acetylcholinesterase inhibitory activity, cytotoxic effect on corneal epithelial cells of sheep, as well as phytochemicals assays and identification of phenolic compounds and organic acids present in the extracts by UPLC - MS. The highest values of total phenolic content were obtained with peel and seed extracts and to ascorbic acid in the seed of siriguela, showing better antioxidant activities of seeds and peel extracts of siriguela and umbu. Lipid peroxidation assays indicated that genipap pulp is a promising antioxidant. Genipap pulp and siriguela seed ethanol extracts presented an acetylcholinesterase inhibition zone similar to that of the positive control, carbachol. The investigation of phenols and organic acid contents revealed the presence of quercetin (48,38 to 3,88 μg.g-1), quinic acid (43,28 to 41,88 μg.g-1) and citric acid (3,78 to 0,43 μg.g-1) in several extracts and chlorogenic acid with the highest amount found in siriguela seeds (356,93 μg.g-1). These data suggest new uses for these foods as potential antioxidant supplements for the food, pharmaceutical and cosmetic industries.
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
A partir das cascas, polpas e sementes do jenipapo (Genipa americana Linnaeus), umbu (Spondia tuberosa Arruda), siriguela (Spondia purpurea Linnaeus) e mangaba (Hancornia speciosa Gomes) foram preparados extratos etanólicos, com exceção da mangaba onde o extrato foi preparado utilizando casca e polpa. Os extratos foram submetidos à investigação da capacidade antioxidante através dos métodos de determinação do conteúdo de ácido ascórbico e fenóis totais, sequestro do radical 2,2-difenil-1-picril-hidrazil e 2,2´- azinobis (3-etilbenzotiazolina-6-ácido sulfônico), capacidade antioxidante de redução do ferro e cobre, lipoperoxidação utilizando um sistema de membranas biomimético e mensuração enzimática da catalase e superóxido dismutase. Também foi avaliada a atividade inibitória da enzima acetilcolinesterase, o efeito citotóxico em células epiteliais da córnea de ovelhas, além da realização de ensaios fitoquímicos e a identificação de compostos fenólicos e ácidos orgânicos presentes nos extratos. Os maiores teores de fenóis totais foram obtidos nos extratos das cascas e sementes e no referente ao ácido ascórbico na semente da siriguela, apresentando melhor atividade antioxidante os extratos das sementes e cascas da siriguela e do umbu. Porém, no ensaio de peroxidação lipídica, o extrato etanólico da polpa de jenipapo demonstrou ser um antioxidante promissor. Os extratos etanólicos da polpa do jenipapo e da semente da siriguela apresentaram uma zona de inibição da enzima acetilcolinesterase semelhantes ao controle positivo, carbacol. Na investigação dos compostos fenólicos e ácidos orgânicos por UPLC-MS verificou-se a presença nos extratos de quercetina (48,38 a 3,88 μg.g-1), ácido quínico (43,28 a 41,88 μg.g-1), ácido cítrico (3,78 a 0,43 μg.g-1) em vários extratos. E ácido clorogênico, na semente da siriguela (356,93 μg.g-1). Os resultados obtidos desses extratos sugerem novas formas de utilização desses alimentos como potenciais suplementos antioxidantes na indústria alimentícia, farmacêutica e cosmética.

Plus de 300 alcaloïdes d’Amaryllidaceae doués d’activités biologiques ont été isolés à partir des plantes appartenant à cette famille. De nos jours, seule la galanthamine, utilisée pour le traitement palliatif de la maladie d’Alzheimer, est commercialisée. L’accumulation de ces alcaloïdes dans les plantes est limitée. La culture in vitro est une méthode alternative intéressante pour l’obtention plus aisée de ces alcaloïdes à haute valeur ajoutée. Le premier objectif de ce travail vise à développer une méthode de purification efficace, simple et rapide des extraits de plantes préalablement à leur analyse en LCMS et GCMS. Le second objectif est d’étudier l’effet de plusieurs facteurs exogènes, ajoutés au milieu de culture de bulbilles de Leucojum aestivum et de sa variété Gravety Giant en bioréacteurs RITA®, sur les voies de biosynthèse de la galanthamine et de la lycorine. La variation des paramètres exogènes a permis une accumulation accrue en galanthamine et en lycorine (0,814 mg/g et 1,54 mg/g de matière sèche respectivement) dans les bulbilles. Le troisième objectif porte sur l’isolement et l’identification d’endophytes à partir de bulbes in vivo et in vitro de trois espèces d’Amaryllidaceae (L. aestivum, Narcissus pseudonarcissus et Galanthus elwesii). Des bactéries endophytes du genre Bacillus ont été identifiées. Un nouvel alcaloïde a été isolé à partir des cultures bactériennes
Over 300 Amaryllidaceae alkaloids possessing a wide range of biological activities have been isolated from plants belonging to this family. Galanthamine, used for the palliative treatment of Alzheimer’s disease, is the only one commercialized. The biodisponiblity of these alkaloids is low. In vitro culture offers an alternative yet interesting approach for the biotechnological production of these valuable alkaloids. The aim of this work was, first, to develop a fast, efficient and easy purification method of plant extracts prior to their phytochemical analysis both in LCMS and GCMS. Second, the combined effects of bioreactor RITA® culture and feeding with different exogenous factors on the biosynthetic pathway of both galanthamine and lycorine were studied. The experiments were conducted both with Leucojum aestivum and L. aestivum ‘Gravety Giant’ bulblets. The variation of several exogenous parameters resulted in a better accumulation of galanthamine and lycorine (0.814 mg/g and 1.54 mg/g dry weight respectively) in the bulblets. The third aim was to isolate and identify alkaloid producing endophytes from in vivo and in vitro bulbs of three Amaryllidaceae species (L. aestivum, Narcissus pseudonarcissus and Galanthus elwesii). Bacterial endophtes belonging to the Bacillus genus were identified. A new alkaloid was isolated from bacterial liquid cultures

There are concerns about using synthetic phenolic antioxidants such as butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) as food additives because negative effects to human health have been reported. thus, a replacement of these synthetics by antioxidant extractions from various foods has been proposed. By the generation of more than eight thousand different phenolics, fruits and vegetables are the prime sources of natural antioxidants. The consumption of fruits and vegetables has been strongly linked with several health benefits, a result of their medicinal properties and high nutritional value. Antioxidants control and reduce the oxidative damage in foods by delaying or inhibiting oxidation caused by reactive oxygen species (ROHs); ultimately increasing the shelf-life and quality of these foods.Beta-carotene, ascorbic acid, and many phenolics play dynamic roles in delaying aging, reducing inflammations, and preventing certain cancers. Increasing the consumption of fruits and vegetables has been recommended by many agencies and health care systems. In order to extract, measure, and identify bioactive compounds from a wide variety of fruits and vegetables, researchers use many different techniques and methods. Ultrasonic assisted technique (UAE) is known to be one of the easiest extraction techniques because it uses common laboratory equipment such as ultrasonic baths and probes. In this technique, a smashed sample is mixed with the suitable solvent and placed into the ultrasonic bath while temperature and extraction time are controlled. Five experiments were conducted to study the effects of different ultrasonic parameters on the polyphenol and antioxidant content and antimicrobial activity of extracts from plants grown in southern Illinois. In the first experiment, ultrasonic treatments at different frequencies,temperatures, power levels, and exposure times were compared for their effects on the yield of total phenolics, total flavonoids and antioxidant activities of spinach extracts. The best conditions for extraction judged by yields were an ultrasonic frequency of 37 kHz, extraction time of 30 min, reaction temperature of 400C, and ultrasonic power of 50%. The mean yield (mg/100g), total phenolics (mg gallic acid equivalence/ g DW), flavonoids (mg/g DW), % DPPH free radical scavenging activity, and % ferric reducing antioxidant power were all high (64.88±21.84, 33.96±11.30, 27.37±11.85 , 64.18±16.69 and 70.25 ±9.68). In the second experiment, the study was designed to optimize ultrasonic-assisted extraction of lutein and β-carotene from spinach. Further, the identities and purity of the natural products in the TLC spots were assayed using matrix-assisted laser desorption/ionization time-off-light mass spectrometry (MALDI-TOF MS) analysis. The optimal UAE settings for output of lutein and β-carotene simultaneously from spinach extracts were an extraction temperature of 40 °C, extraction power of 40% (28 W/cm3) and extraction time of 16 min. UAE assisted extraction of carotenes from spinach can provide a source of lutein and β-carotene for the dietary supplement industry. In the third study, investigations of the effects of extraction parameters for UAE on antioxidant activity of peach and pumpkin extracts; and studying the chemical structures of samples before and after processing by Fourier transformed infrared spectroscopy were made. The optimal conditions for peach extracts were an extraction temperature of 41.53 ̊C, power of 43.99 % and time of 27.86 min for total phenolics. The optimal conditions for extractions of total phenolics from pumpkins were inferred to be a temperature of 41.45 ̊C, a power of 44.60 % and a time of 25.67 min. In the fourth study, TLC followed by Quantity-OneTM (Biorad) image analysis was used as a simple and rapid method for identification and quantification of compounds in complex mixtures. The results were consistent under optimal conditions among the experimental values and their predicted values. The MALDI-TOF MS technique was also applied for result confirmation. In the last experiments, UAE and Box-Wilson design were applied to optimize the extraction of antimicrobial activities from spinach extracts and to test their antimicrobial activities on both Gram-negative and Gram-positive bacteria. The spinach extracts exhibited antimicrobial activities against both bacterial pathogens with MICs in the 60-100 mg.ml-1 range. SEM showed that cells of the treated bacterial strains were damaged and showed structural defects compared to the controls. RAPD analysis of genomic DNA showed that the number and sizes of amplicons were decreased for the treated pathogens compared with non-treated strains.

Thesis (Ph.D.) - University of Glasgow, 2008.
Ph.D. thesis submitted to the Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, 2008. Includes bibliographical references. Print version also available.

A tropical shrub from Euphorbiaceae (Juss.) family, namely Acalypha wilkesiana var. macafeana hort. was investigated for its antioxidant and antibacterial properties. The antioxidant properties were assayed by Ferric Reducing Antioxidant Power (FRAP) assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and β-carotene bleaching assay. Assessment of its antibacterial properties was conducted with pour plate disc diffusion assay and dilution methods. The plant was collected, dried, grinded and soaked continuously in four different solvent starting from non-polar to polar solvent: hexane, ethyl acetate, ethanol and water. The crude extracts were concentrated under pressure and kept in -20 °C prior to investigation. The ethanol extract of A. wilkesiana var. macafeana hort. exhibited good antioxidant and antibacterial activities with results more potent than the standards used. To further locate the bioactive constituents of the plant, we fractionated the ethanol extract leading to five fractions, namely F1, F2, F3, F4 and F5. Both antioxidant and antibacterial assays were conducted upon all the five fractions. Results showed profound activity from F5 of both antioxidant and antibacterial properties which warrants further investigation. To shed light on the active constituents of F5, identification was done with high performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LCMS) and nuclear magnetic resonance (NMR). Separation was obtained with reversed phase HPLC which showed one major compound and 6 minor compounds. The major compound was collected with a fraction collector and identified as geraniin via interpretation and comparison of its NMR shifts, while the other 3 compounds were identified by fragmentations of LC-MS. The compounds identified are β-glucogallin, potentillin and sanguiin H-6. All identified compounds are ellagitannins, except for β-glucogallin which is a gallotannin. The in vitro cell-based assay was performed to HepG2 cells to assess the ability of antioxidants like ellagitannins to protect cells against oxidative insults, and F5 was observed to be able to protect cells against cell death induced by t-BHP insults in a dose-dependent manner. F5 was also found non-toxic in the concentration needed to protect the cells, which is 100 µg/mL. We then explored the synergistic property of the tannin fraction, F5 with commercial antibiotics and observed F5 and ampicillin inhibit the growth of Staphylococcus aureus synergistically. Field Emission Scanning Electron Microscopy (FESEM) analysis was able to demonstrate that the bactericidal mechanism of F5 involves cell wall lysis as the result illustrates indentation of the cell surface and some showed total collapse of the cells. To explore its ability to be formulated and used as a topical agent for treating bacterial infections, a preliminary formulation was made incorporating F5, and formulated with 3 different bases. The formulation made with the paraffin base was observed able to exert the antibacterial property of F5 against Staphylococcus aureus in the in-vitro assay. In vivo animal study on guinea pigs with an incised cut infected with Staphylococcus aureus and treated with the formulation showed that the closure and healing of the wound was faster than Burnol®. Our results indicate possible use of ellagitannins from A. wilkesiana var. macafeana hort. with ampicillin to treat Staphylococcus aureus infections as it is bactericidal via a mechanism involving cell lysis. It also illustrates the possibility to be used as a topical wound healing agent with respect to its antibacterial and antioxidant properties. Ellagitannins from A. wilkesiana var. macafeana hort. can be viewed as a possible bactericidal agent that can contribute to the development of topical antibacterial drug or cosmetics in tropical countries.

This thesis investigated the integration of pharmacological and phytochemical data of medicinal plants from the Cree of Eeyou Istchee in Northern Quebec. Data from these 17 plant extracts were assessed for patterns of biological activity and chemical signals that could be explained by taxonomic or plant organ groupings. The Squamish medicinal plant Oplopanax horridus (Sm.) Miq. was also assessed for enzyme inhibition activity across multiple extracts and for bioactive compounds using an untargeted metabolomics approach. A comprehensive data set was assembled documenting the relative activities on the 17 plant extracts in 69 cell-free and cell-based bioassays covering activity on glucohomeostasis, effects of hyperglycemia, and capacity for enzyme inhibition. Multivariate analysis suggests that the leaf part extracts are particularly associated with antioxidant and antiglycation activities, while another discrete group of extracts associate strongly with other sets of glucohomeostasis assays. The activity of extracts on enzyme inhibition appears to be the factor most strongly driving the majority of activity patterns, likely because extracts that interact strongly with more metabolic enzymes will have more effects on other targets in the body. The phytochemical profiles of the Cree medicinal plants were assessed in two ways. First, spectroscopic and chromatographic data for the plant extracts was compared to a database of phytochemical standards using a proprietary Waters software, UNIFI, to match known signals of chemical standards to unidentified peaks in the plant extracts. Second, similarly collected spectroscopic data for the Cree plant extracts was processed using the software MZMine for multivariate analysis in R, revealing the chemical diversity of the bark extracts in relation to the fruit and leaf extracts. Additionally, marker signals were determined for major sample groupings, and the capacity for this analytical approach to be used to tentatively identify unique compounds was demonstrated. Through bioassay guided fractionation of the O. horridus inner bark extract using the CYP 3A4 inhibition assay, the DCM subfraction midway through the non-polar elution on open column chromatography was determined to be the most potent. This fraction contained 10 major peaks on HPLC-DAD analysis. The hot water extract was found to have negligible activity on CYP 3A4 inhibition. Together, this research provides the first integrated look at the pharmacological and phytochemical data from across the Cree anti-diabetic medicinal plants in a statistical way, as well as providing a first look at O. horridus for inclusion in the anti-diabetes project.

CAPES; Fundação Araucária
A espécie Moringa oleifera (Moringaceae) é uma planta que possui ampla aplicação industrial, alto valor nutricional e que, além disso, também exibe diversas atividades biológicas. Utilizadas na medicina popular, as folhas de M. oleifera já demonstraram possuir grande variedade de moléculas bioativas, inclusive compostos fenólicos, os quais são, possivelmente, os responsáveis pelo potencial antioxidante desta parte da planta. Apesar do crescente interesse sobre a espécie e, especificamente, sobre o seu potencial fitoquímico, são escassos os trabalhos que relatam o isolamento e a identificação dos compostos bioativos presentes nas suas folhas, principalmente em exemplares cultivados no Brasil. Sendo assim, os objetivos deste trabalho foram comparar dois métodos de extração de compostos bioativos e, na sequência, isolar bioguiadamente compostos com atividade antioxidante das folhas de M. oleifera coletadas no município de Itajaí (Santa Catarina, Brasil). O monitoramento bioguiado foi realizado com ensaios in vitro de determinação da atividade antioxidante: capacidade de redução do reagente Folin-Ciocalteau, FRAP, sequestro dos radicais DPPH e ABTS, além do ORAC. A técnica de CLAE-DAD foi utilizada para a caracterização química e acompanhamento das etapas do isolamento. A principal diferença prática entre os métodos de extração avaliados foi o preparo de um extrato hidroalcoólico inicial, no processo de extração 1. A partir dos resultados de determinação da atividade antioxidante, interpretados com o auxílio de ferramentas estatísticas (teste de Tukey e teste t pareado), foi possível perceber que o potencial das folhas de M. oleifera sofreu variações em função da forma de extração e dos solventes utilizados. Em geral, as frações produzidas a partir do processo de extração 1 apresentaram maior atividade antioxidante e perfis cromatográficos com sinais mais intensos. Com base nestes resultados, a fração obtida com acetato de etila, no processo de extração 1, foi selecionada para dar sequência ao isolamento bioguiado. A purificação desta fração em coluna aberta preenchida com sílica gel gerou 61 subfrações, as quais, após análise de CCD, foram agrupadas em 18. A avaliação da atividade antioxidante das subfrações agrupadas mostrou que cinco apresentavam grande potencial. Contudo, em função do rendimento, apenas três puderam dar sequência ao isolamento. Nesta etapa, uma análise adicional foi realizada: a determinação da atividade antioxidante por CLAE on-line com o ABTS•+, que permitiu definir quais dos compostos presentes nas três subfrações possuíam maior potencial e, por isso, seriam isolados. Desta forma, cinco compostos foram isolados pela técnica de CLAE semipreparativa, sendo que dois foram testados frente ao ensaio de sequestro do DPPH•. Os valores de EC50 obtidos, 30,34 e 38,72 μg/mL, estão próximos aos encontrados na literatura para substâncias isoladas de outras matrizes naturais. A técnica de RMN permitiu identificar um flavonol glicosilado. Os resultados deste trabalho mostraram que as folhas de M. oleifera coletadas em Itajaí são fonte de compostos fenólicos com potencial antioxidante e, por isso, promissoras para aplicação nas indústrias de cosméticos, alimentos e farmacêutica.
Moringa oleifera (Moringaceae) is a plant that has wide industrial application, high nutritional value and, also, exhibits several biological activities. Used in folk medicine, M. oleifera leaves have already been shown to possess a wide variety of bioactive molecules, including phenolic componds, which are possibly responsible for the antioxidant potential of this part of the plant. Despite the growing interest in this species and, specifically, in its phytochemical potential, there are few studies about the isolation and identification of bioactive compounds present in M. oleifera leaves, especially in specimens grown in Brazil. Therefore, the aims of this work were to compare two methods for extracting bioactive componds and, than, to isolate compounds with antioxidant activity of M. oleifera leaves collected in Itajaí (Santa Catarina, Brazil) by a bioguided study. The bioguided monitoring was carried out with in vitro assays to determine the antioxidant activity: Folin-Ciocalteau reagent reduction capacity, FRAP assay, DPPH and ABTS radical scavenging methods and, also, the ORAC assay. HPLC-DAD technique was used for chemical characterization and monitoring of the isolation stages. The main practical difference between the evaluated extraction methods was the preparation of an initial hydroalcoholic extract, in the extraction process 1. From the results of the antioxidant activity determination, interpreted with the aid of statistical tools (Tukey’s test and paired t-test), it was possible to see that the potential of M. oleifera leaves varied depending on the extraction form and on the solvents used. In general, the fractions prepared from the extraction process 1 showed higher antioxidant activity and chromatographic profiles with more intense signals. Based on these results, the fraction obtained with ethyl acetate, in the extraction process 1, was selected for the bioguided isolation. The purification of this fraction on an open column of silica gel generated 61 subfractions, which, after TLC analysis, were grouped in 18. The evaluation of the antioxidant activity of grouped subfractions showed that five of them presented great potential. However, depending on the yield, only three could follow the isolation. In this step, an additional analysis was performed: the determination of the antioxidant activity by an on-line HPLC method with the ABTS•+. This technique allowed defining which of the compounds presented in each subfraction had higher potential and, therefore, would be isolated. In this way, five compounds were isolated by semipreparative HPLC, two of them were tested by the DPPH• scavenging assay. The obtained EC50 values, 30.34 and 38.72 μg/mL, are close to those found in literature for substances isolated from other natural matrices. The NMR technique allowed identifying a flavonol glucoside. The results of this work showed that M. oleifera leaves collected in Itajaí are source of phenolic compounds with antioxidant potential and, therefore, are promising for the application in cosmetics, food and pharmaceutical industries.

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The apple is a very popular fruit because of its aroma, flavor and attractive look, its consumption is related to several benefits to human health. With industrialization, a residue of juice extraction is generated, the apple pomace, with similar characteristics of nutrients and bioactive compounds that fruit. This apple pomace after stabilization becomes a by-product with excellent biotechnological applications. The positive effects of irradiation in food has been exploration, especially in increasing the useful life of the product. This study aimed to evaluate the effects of gamma radiation on the physicochemical characteristics, thermogravimetric, microstructural and microbiological, phenolic compounds and antioxidant potential of apple cake meal, for nine months of storage, with the help of multivariate techniques such as analysis Principal Components (PCA) and Hierarchical Cluster Analysis (HCA). Gamma radiation and the storage period did not influence the parameters of lipid, sucrose, fructose and ashes. Reducing sugars were not affected by the irradiation and the dose of 1 kGy remained stable during the storage period. The glucose was stable from the third month, and the highest levels were found at a dose of 2 kGy. The mineral calcium levels were higher in the irradiated samples. The dietary fiber content of the irradiated samples resulted in lower values compared to the control sample, but remained within the contents found in the literature. The irradiated samples were stable during storage for protein, lipid, dietary fiber, total reducing sugar, minerals K, Zn, Fe and Mn. The Phlorizin was the phenolic compound that remained constant levels during the search. Chlorogenic acid and its class hydroxycinnamic acids increased proportionally with time in the irradiated samples. The chemometric approach is effective to facilitate the visualization of samples of apple cake meal, the physical-chemical parameters and by HCA, the phenolic compounds were separated into three groups, the first characterized in that only samples without irradiation, the Then, samples with 2 kGy and third represent characterized by a dose of 1 kGy. Thermogravimetry curves with four major mass loss in consecutive reactions. In the hotomicrographs, there was an organic material composition and heterogeneous, with agglomerates of particles which have irregular shapes and sizes. From a microbiological point of view the dose of 2 kGy was lethal to the inhibition of yeasts and molds to the end of the study. This raw material is a great source of fiber and can be applied to products with high added value, aiming economy and reducing the environmental impact.
A maçã é uma fruta muito popular devido ao seu aroma, sabor e visual atrativo, seu consumo está relacionado a vários benefícios a saúde humana. Com a industrialização, é gerado um resíduo da extração do suco, o bagaço de maçã, com características similares de nutrientes e compostos bioativos que a fruta. Esse bagaço quando estabilizado se torna um subproduto com excelentes aplicações biotecnológicas. Os efeitos positivos da irradiação em alimentos tem sido exploratório, principalmente no aumento do tempo de vida útil do produto. Este trabalho teve por objetivo avaliar os efeitos da radiação gama nas características físico-químicas, termogravimétricas, microestruturais e microbiológicas, compostos fenólicos e potencial antioxidante da farinha de bagaço de maçã, durante nove meses de armazenamento, com auxílio de técnicas multivariadas, como a Análise de Componentes Principais (PCA) e a Análise Hierárquica de Agrupamentos (HCA). A radiação gama e o período de armazenamento não influenciaram nos parâmetros de lipídeos, sacarose, frutose e cinzas. Os açúcares redutores não foram influenciados pela irradiação e com a dose de 1 kGy se manteve estável durante o período de armazenamento. A glucose se manteve estável a partir do terceiro mês, e os maiores teores foram encontrados com a dose de 2 kGy. O mineral cálcio apresentou maiores teores nas amostras irradiadas. Os teores de fibra alimentar nas amostras irradiadas resultam em menores valores comparadas a amostra controle, mas continuaram dentro dos índices encontrados na literatura. As amostras irradiadas durante o armazenamento foram estáveis para os teores de proteína, lipídeos, fibra alimentar, açúcar redutor total, os minerais K, Zn, Fe e Mn. A floredizina foi o composto fenólico que manteve teores constantes durante a pesquisa. O ácido clorogênico e a sua classe de ácidos hidroxicinâmicos aumentaram proporcionamente com o tempo nas amostras irradiadas. A abordagem quimiométrica se mostrou eficaz ao facilitar a visualização das amostras da farinha de bagaço de maçã, nos parâmetros físicoquímicos e através da HCA, os compostos fenólicos foram separados em 3 grupos, o primeiro caracterizado por apresentar apenas as amostras sem irradiação, o segundo, amostras com 2 kGy e o terceiro caracterizou-se por representar a dose de 1 kGy. As curvas de termogravimetria apresentam quatro perdas principais de massa, em reações consecutivas. Com as fotomicrografias, observou-se um material de composição orgânica, bem heterogêneo, com aglomerados de partículas que apresentam formas e tamanhos irregulares. Do ponto de vista microbiológico a dose de 2 kGy foi letal para a inibição de bolores e leveduras ao término da pesquisa. Essa matéria prima é uma ótima fonte de fibras e pode ser aplicada em produtos com alto valor agregado, visando economia e reduzindo o impacto ambiental.

Thesis submitted in fulfilment of the requirements for the degree Master of Technology: Consumer Science: Food and Nutrition in the Faculty of Applied Sciences at the Cape Peninsula University of Technology 2012
The chronic diseases of lifestyle, tuberculosis and Human immunodeficiency virus/Acquired immunodeficiency syndrome have a high prevalence in South Africa. These diseases are characterised by oxidative stress and a chronic inflammatory state that contribute to both the development and the acceleration of these diseases. Research into the phytochemical plant food components suggest that these substances could possibly play a vital role in the prevention of such disease. Corn steep liquor (CSL) is a waste product with an exceptionally high polyphenol content and total antioxidant capacity (TAC). This led to the suggestion that it could be utilised in the development of food products with functional food characteristics. Stock cubes, due to the widespread use among consumers of different socio-economic backgrounds, were identified as vehicle for the delivery of the CSL with its phytochemical content. This lead to the development of a stock cube utilising CSL as a source of phytochemical polyphenolic antioxidants with the micronutrients zinc, selenium and copper as added support to immunonutrition, along with iron due to the wide spread prevalence of iron deficiency in the South African population. The acceptability of the developed stock cube was tested by preparing savoury rice and pea soup and having blue collar (n = 50) and white collar (n = 49) participants rate the acceptability on a 9-point hedonic scale, ranging from “dislike extremely” to “like extremely”. The savoury rice received a 41% “like very much” rating followed by a 24% “like moderately” rating. The pea soup was rated even more positively as it received a 42% “like very much” rating and a 29% “like extremely” rating. In addition, among the blue collar participants, significant (p < 0.05 for each) findings occurred with a greater liking of the sample dishes prepared with the developed stock cube by participants of the ethnic Black grouping, with Xhosa as home language and being married, as well as those participants who habitually prepare the meals in the household. A significant (p < 0.05) finding with the white collar participants was a greater liking of the sample dishes prepared with the developed stock cube among those participants who would be willing to make use of a stock cube with health benefits. The developed stock cube was also subjected to six months accelerated shelf-life stability testing, reflecting twelve months real time storage. The parameters measured were microbial growth, oxidative rancidity, antioxidant status and organoleptic changes. All microbial growths tested remained within the acceptable specification ranges. Oxidative rancidity, measured as the peroxide value, was detected analytically at the fifth and sixth samplings, but was not detectable organoleptically. The antioxidant status, measured as the TAC and the total polyphenol content, remained relatively stable during the testing period. Though no noticeable organoleptic changes were observed during the stability testing, there was a darkening of the colour by the second sampling. The results of the acceptance testing, as well as the shelf-life stability testing, support the conclusions that the study objectives of developing a stock cube with functional food characteristics and it being received positively by consumers were achieved. Keywords: stock cubes; corn steep liquor; phytochemicals; consumer acceptance; new product development

Woo Ka-yan.
"September 2003."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references.
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 Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.