Phytochemical analyses and Brine shrimp (Artemia Salina) lethality studies on Syzygium cordatum
- Authors: Chiguvare, Herbert
- Date: 2013
- Subjects: Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11336 , http://hdl.handle.net/10353/d1004352 , Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Description: 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.
- Full Text:
- Date Issued: 2013
- Authors: Chiguvare, Herbert
- Date: 2013
- Subjects: Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11336 , http://hdl.handle.net/10353/d1004352 , Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Description: 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.
- Full Text:
- Date Issued: 2013
Beta-N-methylamino-L-alanine in South African fresh water cyanobacteria : incidence, prevalence, ecotoxicological considerations and human exposure risk
- Authors: Esterhuizen-Londt, Maranda
- Date: 2010
- Subjects: Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10306 , http://hdl.handle.net/10948/1473 , Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Description: β-N-methylamino-L-alanine (BMAA) is a non-proteinogenic amino acid associated with human neurodegenerative disease. Due to the cosmopolitan nature of cyanobacteria, detection of BMAA in cyanobacteria has caused concerns about human exposure risk. This study was therefore based on the hypothesis that BMAA poses a health risk to humans either by direct ingestion or by indirect exposure to BMAA from a cyanobacterial source via a freshwater food chain. A validated gas chromatography-mass spectrometry (GC-MS) BMAA analysis method and a confirmatory liquid chromatography-mass spectrometry (LC-MS) method, with improved sensitivity, were developed in addition to a LC-MS/MS method for analyte confirmation. These methods were used to quantify BMAA in South African cyanobacteria, isolated from various potable water reservoirs. The majority of the isolates tested, contained BMAA. Possible human exposure by direct consumption of BMAA released from cyanobacterial blooms was investigated by the development of a robust solid phase extraction (SPE) method used for BMAA concentration and quantification in raw and treated tap water. Despite the use of the SPE method that facilitated the concentration of BMAA from large quantities of water, no free dissolved BMAA was detected in raw or processed fresh water. The fate of exogenous BMAA was therefore investigated firstly by evaluating the efficacy of standard water treatment processes employed in South Africa and secondly by investigating the possibility of BMAA bioaccumulation and biomagnification in aquatic food chains. Standard water treatment processes proved highly efficient at removing free dissolved BMAA, explaining the absence of BMAA in treated tap water. However, the cause of the BMAA absence in raw potable water remained unknown. Uptake of BMAA by model aquatic organisms was investigated in controlled experiments. BMAA uptake was documented in both Ceratophyllum demersum and Daphnia magna, however, BMAA-protein association and biomagnification were not observed in D. magna. BMAA had an inhibitory effect on the oxidative stress enzyme acitivties of both organisms tested (as well as human S9 extracts), resulting in accumulation of detrimental reactive oxygen species (ROS) in the cells. Exposure of crop plants to BMAA in controlled experiments resulted in BMAA uptake, protein association, and subsequent inhibition of the antioxidative enzyme activities. However, BMAA was detected in neither free nor protein-associated form in natural crop plants irrigated with known BMAA-containing bloom water. Post-mortem liver samples of Clarias gariepinus (Catfish) and Crocodylus niloticus (Crocodile), from a natural fresh water ecosystem that experienced frequent cyanobacterial blooms, contained both free and protein-associated BMAA. Higher BMAA concentrations were found in crocodile liver samples compared to fish liver samples, strongly suggesting biomagnification from one trophic level to the next. BMAA concentrations corresponded to crocodile age. This is the first report of bioaccumulation and biomagnification in two trophic levels in a fresh water ecosystem. These findings strongly suggest possible human exposure via aquatic food chains of cyanobacterial origin. Direct BMAA exposure via drinking water is not plausible due to the efficiency of standard water treatment processes to remove BMAA. The use of raw water for agricultural and recreational use, however, remains a problem. The development of management strategies as well as daily tolerable levels for BMAA is urgently required.
- Full Text:
- Date Issued: 2010
- Authors: Esterhuizen-Londt, Maranda
- Date: 2010
- Subjects: Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10306 , http://hdl.handle.net/10948/1473 , Cyanobacteria , Bioaccumulation , Chromatographic analysis , Neurotoxic agents
- Description: β-N-methylamino-L-alanine (BMAA) is a non-proteinogenic amino acid associated with human neurodegenerative disease. Due to the cosmopolitan nature of cyanobacteria, detection of BMAA in cyanobacteria has caused concerns about human exposure risk. This study was therefore based on the hypothesis that BMAA poses a health risk to humans either by direct ingestion or by indirect exposure to BMAA from a cyanobacterial source via a freshwater food chain. A validated gas chromatography-mass spectrometry (GC-MS) BMAA analysis method and a confirmatory liquid chromatography-mass spectrometry (LC-MS) method, with improved sensitivity, were developed in addition to a LC-MS/MS method for analyte confirmation. These methods were used to quantify BMAA in South African cyanobacteria, isolated from various potable water reservoirs. The majority of the isolates tested, contained BMAA. Possible human exposure by direct consumption of BMAA released from cyanobacterial blooms was investigated by the development of a robust solid phase extraction (SPE) method used for BMAA concentration and quantification in raw and treated tap water. Despite the use of the SPE method that facilitated the concentration of BMAA from large quantities of water, no free dissolved BMAA was detected in raw or processed fresh water. The fate of exogenous BMAA was therefore investigated firstly by evaluating the efficacy of standard water treatment processes employed in South Africa and secondly by investigating the possibility of BMAA bioaccumulation and biomagnification in aquatic food chains. Standard water treatment processes proved highly efficient at removing free dissolved BMAA, explaining the absence of BMAA in treated tap water. However, the cause of the BMAA absence in raw potable water remained unknown. Uptake of BMAA by model aquatic organisms was investigated in controlled experiments. BMAA uptake was documented in both Ceratophyllum demersum and Daphnia magna, however, BMAA-protein association and biomagnification were not observed in D. magna. BMAA had an inhibitory effect on the oxidative stress enzyme acitivties of both organisms tested (as well as human S9 extracts), resulting in accumulation of detrimental reactive oxygen species (ROS) in the cells. Exposure of crop plants to BMAA in controlled experiments resulted in BMAA uptake, protein association, and subsequent inhibition of the antioxidative enzyme activities. However, BMAA was detected in neither free nor protein-associated form in natural crop plants irrigated with known BMAA-containing bloom water. Post-mortem liver samples of Clarias gariepinus (Catfish) and Crocodylus niloticus (Crocodile), from a natural fresh water ecosystem that experienced frequent cyanobacterial blooms, contained both free and protein-associated BMAA. Higher BMAA concentrations were found in crocodile liver samples compared to fish liver samples, strongly suggesting biomagnification from one trophic level to the next. BMAA concentrations corresponded to crocodile age. This is the first report of bioaccumulation and biomagnification in two trophic levels in a fresh water ecosystem. These findings strongly suggest possible human exposure via aquatic food chains of cyanobacterial origin. Direct BMAA exposure via drinking water is not plausible due to the efficiency of standard water treatment processes to remove BMAA. The use of raw water for agricultural and recreational use, however, remains a problem. The development of management strategies as well as daily tolerable levels for BMAA is urgently required.
- Full Text:
- Date Issued: 2010
Identification and characterization of novel oncology related platinum complexes using chromatographic and mass spectrometric techniques
- Authors: Wentzel, Mauritz
- Date: 2008
- Subjects: Chromatographic analysis , Spectrum analysis
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10310 , http://hdl.handle.net/10948/715 , Chromatographic analysis , Spectrum analysis
- Description: In this thesis mass spectral and chromatographic techniques were developed and applied to identify and characterise numerous novel platinum(II) and (IV) compounds designed as anticancer agents. In a novel method for the synthesis of cis-oxalato(trans- -1,2- cyclohexanediamine)platinum(II) or oxaliplatin these techniques could be applied to differentiate between the molecular complex and the autoionised analogue (viz. Ptdach2 2+Ptox2 2-). In another novel synthetic method for the same compound the ligand exchange reactions at various temperatures could be investigated and kinetic curves obtained served to illuminate the chemistry involved, indicating the role of small amounts of water in the essentially non-aqueous solvent systems dmf and isoamyl alcohol respectively. These allowed ligand exchange without resulting in hydrolyses even up to 85°C. The ionisation rate of divalent platinum halide complexes was determined for various amine ligands as well as N-S chelate ligands. A comparison of these could suggest why N-S complexes have poor anticancer action. Ionisation was not only studied for neutral molecular species but also for monocationic ones. Relationships could be found with stereochemical aspects of the chelates used. By investigating results of EV-CAD studies thermodynamic data could be obtained which indicated that bond strength decreases from chloro to iodo analogues although extent of ionisation in aqueous solution, i.e kinetic stability, is the reverse. Products formed by the reaction of NO2 gas with Platinum(II) compounds could be identified and separated which greatly contributed to the understanding of the chemistry involved in the formation of mononitro platinum(IV) complexes. Some of these proved to have exceptional anticancer properties. Studies of the interaction of thiol containing biomolecules were performed as a function of time. The results contributed to the understanding of the action of the anticancer agents.
- Full Text:
- Date Issued: 2008
- Authors: Wentzel, Mauritz
- Date: 2008
- Subjects: Chromatographic analysis , Spectrum analysis
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10310 , http://hdl.handle.net/10948/715 , Chromatographic analysis , Spectrum analysis
- Description: In this thesis mass spectral and chromatographic techniques were developed and applied to identify and characterise numerous novel platinum(II) and (IV) compounds designed as anticancer agents. In a novel method for the synthesis of cis-oxalato(trans- -1,2- cyclohexanediamine)platinum(II) or oxaliplatin these techniques could be applied to differentiate between the molecular complex and the autoionised analogue (viz. Ptdach2 2+Ptox2 2-). In another novel synthetic method for the same compound the ligand exchange reactions at various temperatures could be investigated and kinetic curves obtained served to illuminate the chemistry involved, indicating the role of small amounts of water in the essentially non-aqueous solvent systems dmf and isoamyl alcohol respectively. These allowed ligand exchange without resulting in hydrolyses even up to 85°C. The ionisation rate of divalent platinum halide complexes was determined for various amine ligands as well as N-S chelate ligands. A comparison of these could suggest why N-S complexes have poor anticancer action. Ionisation was not only studied for neutral molecular species but also for monocationic ones. Relationships could be found with stereochemical aspects of the chelates used. By investigating results of EV-CAD studies thermodynamic data could be obtained which indicated that bond strength decreases from chloro to iodo analogues although extent of ionisation in aqueous solution, i.e kinetic stability, is the reverse. Products formed by the reaction of NO2 gas with Platinum(II) compounds could be identified and separated which greatly contributed to the understanding of the chemistry involved in the formation of mononitro platinum(IV) complexes. Some of these proved to have exceptional anticancer properties. Studies of the interaction of thiol containing biomolecules were performed as a function of time. The results contributed to the understanding of the action of the anticancer agents.
- Full Text:
- Date Issued: 2008
High performance liquid chromatographic analysis of erythromycin in serum and urine
- Authors: Stubbs, Christopher
- Date: 1985 , 2013-03-13
- Subjects: High performance liquid chromatography , Erythromycin , Erythromycin -- Pharmacokinetics , Chromatographic analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3813 , http://hdl.handle.net/10962/d1004581 , High performance liquid chromatography , Erythromycin , Erythromycin -- Pharmacokinetics , Chromatographic analysis
- Description: Erythromycin, a macrolide antibiotic used mainly against gram-positive bacteria has been in clinical use since 1952 (1). Previous pharmacokinetic data published on this antibiotic have been derived predominantly from microbiological assay techniques. However, these techniques are relatively imprecise as well as being non-specific and extremely tedious to perform. A novel high performance liquid chromatographic analysis of erythromycin in human serum and urine using U.V. detection at 200 nm and/or electrochemical detection using both an amperometric and a coulometric electrochemical detector is presented. The method involves a solid phase extraction procedure followed by a simple phase separation step and chromatography on a reverse phase column. In order to select the optimum U.V. detector for this analysis, five "state of the art" detectors were compared in terms of their signal-to-noise ratios at U.V. wavelengths between 200 and 210 nm. A known metabolite des-N-methylerythromycin is readily detectable using U.V. detection, whilst another metabolite/degradation product anhydroerythromycin is not seen using U.V. detection but is readily observable using an electrochemical detector. The method has a limit of sensitivity of 0.25 μg/mL and 1.00 μg/mL in serum and urine respectively (U.V. detection) and is sufficiently sensitive to monitor serum and urine concentrations of erythromycin in man after administration of a single 500 mg erythromycin stearate tablet. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 1985
- Authors: Stubbs, Christopher
- Date: 1985 , 2013-03-13
- Subjects: High performance liquid chromatography , Erythromycin , Erythromycin -- Pharmacokinetics , Chromatographic analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3813 , http://hdl.handle.net/10962/d1004581 , High performance liquid chromatography , Erythromycin , Erythromycin -- Pharmacokinetics , Chromatographic analysis
- Description: Erythromycin, a macrolide antibiotic used mainly against gram-positive bacteria has been in clinical use since 1952 (1). Previous pharmacokinetic data published on this antibiotic have been derived predominantly from microbiological assay techniques. However, these techniques are relatively imprecise as well as being non-specific and extremely tedious to perform. A novel high performance liquid chromatographic analysis of erythromycin in human serum and urine using U.V. detection at 200 nm and/or electrochemical detection using both an amperometric and a coulometric electrochemical detector is presented. The method involves a solid phase extraction procedure followed by a simple phase separation step and chromatography on a reverse phase column. In order to select the optimum U.V. detector for this analysis, five "state of the art" detectors were compared in terms of their signal-to-noise ratios at U.V. wavelengths between 200 and 210 nm. A known metabolite des-N-methylerythromycin is readily detectable using U.V. detection, whilst another metabolite/degradation product anhydroerythromycin is not seen using U.V. detection but is readily observable using an electrochemical detector. The method has a limit of sensitivity of 0.25 μg/mL and 1.00 μg/mL in serum and urine respectively (U.V. detection) and is sufficiently sensitive to monitor serum and urine concentrations of erythromycin in man after administration of a single 500 mg erythromycin stearate tablet. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 1985
- «
- ‹
- 1
- ›
- »