An investigation into the feasibility of incorporating ketoconazole into solid lipid microparticles
- Authors: Jhundoo, Henusha Devi
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/54701 , vital:26601
- Description: One of the major challenges of the oral administration of ketoconazole (KTZ), an inhibitor of sterol 14α demethylase, used in the management of systemic and topical mycoses in immuno-compromised and paediatric patients is the lack of availability of liquid dosage forms. In order to overcome this challenge, extemporaneous preparations have been manufactured by care-givers and health care providers by crushing or breaking solid oral dosage forms of KTZ and mixing with a vehicle to produce a liquid dosage form that can be swallowed by patients. However, the use of extemporaneous preparations may lead to under or over-dosing if the care-givers are not guided accordingly. Furthermore, the dearth of information on the stability of these KTZ-containing extemporaneous preparations may lead to ineffective antifungal therapy and complicate the problems of resistance as it is difficult to estimate the shelf-lives of these extemporaneous products under varying storage conditions due to the susceptibility of KTZ to chemical degradation. Therefore, there is a need for formulation scientists to develop novel drug delivery systems that avoid the need for extemporaneous preparations, possess well-established limits of stability and minimize the risks of systemic adverse effects to facilitate KTZ therapy. The use of solid lipid microparticles (SLM) as potential carriers for the oral administration of KTZ was investigated since solid lipid carriers are known to exhibit the advantages of traditional colloidal carriers. The research undertaken in these studies aimed to investigate the feasibility of developing and manufacturing solid lipid microparticles (SLM), using a simple micro-emulsion technique, as a carrier for KTZ. Prior to pre-formulation, formulation development and optimization studies of KTZ-loaded SLM, it was necessary to develop and validate an analytical method for the in vitro quantitation and characterization of KTZ in aqueous dispersions of SLM during development and assessment studies. An accurate, precise, specific and sensitive reversed-phase high performance liquid chromatographic (RP-HPLC) method coupled with UV detection at 206 nm was developed, optimized and validated for the analysis of KTZ in formulations. Formulation development studies were preceded by solubility studies of KTZ in different lipids. Labrafil® M2130 CS was found to exhibit the best solubilising potential for KTZ. Pre-formulation studies were also designed to determine the polymorphic behavior and the crystallinity of KTZ and Labrafil® M2130 CS that was used for subsequent manufacture of the solid lipid carriers. DSC and FTIR studies revealed that there were no changes in the crystallinity of KTZ or Labrafil® M2130 CS following exposure to a temperature of 60°C for 1 hour. In addition the potential for physicochemical interaction of KTZ with the lipid Labrafil® M2130 CS was investigated using DSC and FTIR and the results revealed that KTZ was molecularly dispersed in Labrafil® M2130 CS and that it is unlikely that KTZ would interact with the lipid. It was therefore established that KTZ and Labrafil® M2130 CS were thermo-stable at a temperature of 60°C and thus a micro-emulsion technique could be used to manufacture the KTZ-loaded SLM. Drug-free and KTZ-loaded SLM were prepared using a modified micro-emulsion technique that required the use of an Ultra-Turrax® homogenizer set at 24 000 rpm for 5 minutes followed by the use of the Erweka GmbH homogenizer. SLM were characterized in terms of particle size (PS), zeta potential (ZP), shape and surface morphology using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition drug loading capacity (DLC) and encapsulation efficiency (EE) of SLM for KTZ were assessed using RP-HPLC. Formulation development and optimization studies of KTZ-loaded SLM were initially aimed at selecting an emulsifying system that was able to stabilize the SLM in an aqueous dispersion. Successful formulations were selected based on their ability to remain physically stable on the day of manufacture. Pluronic® F68 used in combination with Lutrol® E40, Soluphor® P, Soluplus® produced unstable dispersions on the day of manufacture and these combinations were not investigated further. However, the formulation of a stable KTZ-loaded SLM dispersion was accomplished by use of a combination of Pluronic® F68, Tween 80 and sodium cholate as the surfactant system. Increasing amounts of Labrafil® M2130 CS resulted in the production of particles with low DLC and EE, a large PS and a relatively unchanged ZP. An optimum concentration of 10% w/v Labrafil® M2130 CS was selected to manufacture the KTZ-loaded SLM. Studies to determine the influence of KTZ loading on the quality of SLM revealed that concentrations of KTZ > 5% w/v led to a reduction in DLC and EE and an increase in PS with minimal impact on the ZP. Stability studies conducted at 25°C/65% RH and 40°C/75% RH for up to 30 days following manufacture revealed that batch SLM 15 manufactured using 10% w/v Labrafil® M2130 CS, 5% w/v KTZ and a combination of 4% w/v Pluronic® F-68, 2% w/v Tween 80 and 1% w/v sodium cholate produced the most stable dosage form when stored at 25°C/65% RH for up to 30 days. However, storage at 40°C/75% RH resulted in instability of the formulation. An aqueous dispersion of KTZ-loaded SLM has been developed and assessed and may offer an alternative to extemporaneous preparations used for KTZ therapy in paediatric and immuno-compromised patients.
- Full Text:
- Date Issued: 2015
- Authors: Jhundoo, Henusha Devi
- Date: 2015
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/54701 , vital:26601
- Description: One of the major challenges of the oral administration of ketoconazole (KTZ), an inhibitor of sterol 14α demethylase, used in the management of systemic and topical mycoses in immuno-compromised and paediatric patients is the lack of availability of liquid dosage forms. In order to overcome this challenge, extemporaneous preparations have been manufactured by care-givers and health care providers by crushing or breaking solid oral dosage forms of KTZ and mixing with a vehicle to produce a liquid dosage form that can be swallowed by patients. However, the use of extemporaneous preparations may lead to under or over-dosing if the care-givers are not guided accordingly. Furthermore, the dearth of information on the stability of these KTZ-containing extemporaneous preparations may lead to ineffective antifungal therapy and complicate the problems of resistance as it is difficult to estimate the shelf-lives of these extemporaneous products under varying storage conditions due to the susceptibility of KTZ to chemical degradation. Therefore, there is a need for formulation scientists to develop novel drug delivery systems that avoid the need for extemporaneous preparations, possess well-established limits of stability and minimize the risks of systemic adverse effects to facilitate KTZ therapy. The use of solid lipid microparticles (SLM) as potential carriers for the oral administration of KTZ was investigated since solid lipid carriers are known to exhibit the advantages of traditional colloidal carriers. The research undertaken in these studies aimed to investigate the feasibility of developing and manufacturing solid lipid microparticles (SLM), using a simple micro-emulsion technique, as a carrier for KTZ. Prior to pre-formulation, formulation development and optimization studies of KTZ-loaded SLM, it was necessary to develop and validate an analytical method for the in vitro quantitation and characterization of KTZ in aqueous dispersions of SLM during development and assessment studies. An accurate, precise, specific and sensitive reversed-phase high performance liquid chromatographic (RP-HPLC) method coupled with UV detection at 206 nm was developed, optimized and validated for the analysis of KTZ in formulations. Formulation development studies were preceded by solubility studies of KTZ in different lipids. Labrafil® M2130 CS was found to exhibit the best solubilising potential for KTZ. Pre-formulation studies were also designed to determine the polymorphic behavior and the crystallinity of KTZ and Labrafil® M2130 CS that was used for subsequent manufacture of the solid lipid carriers. DSC and FTIR studies revealed that there were no changes in the crystallinity of KTZ or Labrafil® M2130 CS following exposure to a temperature of 60°C for 1 hour. In addition the potential for physicochemical interaction of KTZ with the lipid Labrafil® M2130 CS was investigated using DSC and FTIR and the results revealed that KTZ was molecularly dispersed in Labrafil® M2130 CS and that it is unlikely that KTZ would interact with the lipid. It was therefore established that KTZ and Labrafil® M2130 CS were thermo-stable at a temperature of 60°C and thus a micro-emulsion technique could be used to manufacture the KTZ-loaded SLM. Drug-free and KTZ-loaded SLM were prepared using a modified micro-emulsion technique that required the use of an Ultra-Turrax® homogenizer set at 24 000 rpm for 5 minutes followed by the use of the Erweka GmbH homogenizer. SLM were characterized in terms of particle size (PS), zeta potential (ZP), shape and surface morphology using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition drug loading capacity (DLC) and encapsulation efficiency (EE) of SLM for KTZ were assessed using RP-HPLC. Formulation development and optimization studies of KTZ-loaded SLM were initially aimed at selecting an emulsifying system that was able to stabilize the SLM in an aqueous dispersion. Successful formulations were selected based on their ability to remain physically stable on the day of manufacture. Pluronic® F68 used in combination with Lutrol® E40, Soluphor® P, Soluplus® produced unstable dispersions on the day of manufacture and these combinations were not investigated further. However, the formulation of a stable KTZ-loaded SLM dispersion was accomplished by use of a combination of Pluronic® F68, Tween 80 and sodium cholate as the surfactant system. Increasing amounts of Labrafil® M2130 CS resulted in the production of particles with low DLC and EE, a large PS and a relatively unchanged ZP. An optimum concentration of 10% w/v Labrafil® M2130 CS was selected to manufacture the KTZ-loaded SLM. Studies to determine the influence of KTZ loading on the quality of SLM revealed that concentrations of KTZ > 5% w/v led to a reduction in DLC and EE and an increase in PS with minimal impact on the ZP. Stability studies conducted at 25°C/65% RH and 40°C/75% RH for up to 30 days following manufacture revealed that batch SLM 15 manufactured using 10% w/v Labrafil® M2130 CS, 5% w/v KTZ and a combination of 4% w/v Pluronic® F-68, 2% w/v Tween 80 and 1% w/v sodium cholate produced the most stable dosage form when stored at 25°C/65% RH for up to 30 days. However, storage at 40°C/75% RH resulted in instability of the formulation. An aqueous dispersion of KTZ-loaded SLM has been developed and assessed and may offer an alternative to extemporaneous preparations used for KTZ therapy in paediatric and immuno-compromised patients.
- Full Text:
- Date Issued: 2015
An investigation into the neuroprotective effects of melatonin in a model of rotenone-induced neurodegeneration
- Authors: Kadanthode, Rubina John
- Date: 2004
- Subjects: Melatonin , Nervous system -- Degeneration -- Treatment , Rotenone
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3763 , http://hdl.handle.net/10962/d1003241 , Melatonin , Nervous system -- Degeneration -- Treatment , Rotenone
- Description: Parkinson’s disease, one of the most common neurodegenerative disorders associated with ageing, is characterised by abnormal and profound loss of nigrostriatal dopaminergic neurons. The cause of Parkinson’s disease is unknown, but epidemiological studies suggest an association with pesticides and other environmental toxins, and biochemical studies implicate oxidative damage and mitochondrial impairment, particularly at the level of complex I enzyme. Recently, rotenone, a commonly used organic pesticide and a classical inhibitor of mitochondrial complex I has been reported to reproduce the specific features of Parkinson’s disease in rodents. The mitochondrial respiratory chain is one of the most important sites of reactive oxygen species production under physiological conditions. Toxic free radicals have been implicated in a variety of neurodegenerative diseases as well as ageing itself. Melatonin, a secretory product of the pineal gland is a multifaceted free radical scavenger and natural antioxidant. In the present study, the neuroprotective effects of melatonin against the environmental neurotoxin, rotenone was investigated. Initial studies showed that inhibition of mitochondrial complex I enzyme by rotenone induced superoxide radical generation. Melatonin, administered to the rat in vivo and in vitro was able to offer neuroprotection by curtailing the production of superoxide radicals induced by rotenone. Mitochondria, being the major target of rotenone, the effects of melatonin were investigated at the mitochondrial level. Melatonin was able to increase the electron transport chain activity thus preventing the respiratory inhibition by rotenone. The pineal hormone also counteracted the action of rotenone on complex I enzyme. These results suggest melatonin’s ability to potentially limit the free radical generation and thereby modulate the mitochondrial functions. The detection and measurement of lipid peroxidation is the evidence most frequently cited to support the involvement of free radical reactions in toxicology and in human disease. Melatonin also offered significant protection in vivo and in vitro against rotenone induced lipid peroxidation. Since iron plays a major role in oxidative damage and in the progression of Parkinson’s disease, the effect of melatonin on both rotenone and iron induced lipid peroxidation was investigated, the results of which show that melatonin affords protection and this was suggested to be due to its interaction with the rotenone-iron complex that might have formed. Electrochemical studies were further used to characterise the interactions between melatonin, rotenone and iron (III). Melatonin was shown to bind with iron and thus reducing their toxicity. Histological studies were undertaken to assess the effects of melatonin on rotenone induced toxicity on the dopaminergic neurons in the rat brain. Rotenone treated brains showed extensive neuronal damage whereas with melatonin less damage was observed. Rotenone induces apoptosis via reactive oxygen species production and apoptotic cell death has been identified in PD brains. Furthermore, the apoptotic cell death was detected and quantified by the TUNEL staining. Rotenone treated sections showed signs of apoptosis whereas with melatonin, less apoptotic damage was observed. The findings of this study indicate that the neurohormone, melatonin may protect against rotenone-induced neurodegeneration. Since melatonin production falls substantially during ageing, the loss of this antioxidant is theorized to be instrumental in the degenerative processes associated with advanced age. Considering how devastating diseases such as Parkinson’s disease, are to a patient and the patient’s families, the discovery of protective agents are a matter of urgency. Further investigations using the pesticide model will help to determine the involvement of environmental exposure in the pathogenesis of human diseases as well as to test therapeutic strategies for the treatment of such diseases.
- Full Text:
- Date Issued: 2004
- Authors: Kadanthode, Rubina John
- Date: 2004
- Subjects: Melatonin , Nervous system -- Degeneration -- Treatment , Rotenone
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3763 , http://hdl.handle.net/10962/d1003241 , Melatonin , Nervous system -- Degeneration -- Treatment , Rotenone
- Description: Parkinson’s disease, one of the most common neurodegenerative disorders associated with ageing, is characterised by abnormal and profound loss of nigrostriatal dopaminergic neurons. The cause of Parkinson’s disease is unknown, but epidemiological studies suggest an association with pesticides and other environmental toxins, and biochemical studies implicate oxidative damage and mitochondrial impairment, particularly at the level of complex I enzyme. Recently, rotenone, a commonly used organic pesticide and a classical inhibitor of mitochondrial complex I has been reported to reproduce the specific features of Parkinson’s disease in rodents. The mitochondrial respiratory chain is one of the most important sites of reactive oxygen species production under physiological conditions. Toxic free radicals have been implicated in a variety of neurodegenerative diseases as well as ageing itself. Melatonin, a secretory product of the pineal gland is a multifaceted free radical scavenger and natural antioxidant. In the present study, the neuroprotective effects of melatonin against the environmental neurotoxin, rotenone was investigated. Initial studies showed that inhibition of mitochondrial complex I enzyme by rotenone induced superoxide radical generation. Melatonin, administered to the rat in vivo and in vitro was able to offer neuroprotection by curtailing the production of superoxide radicals induced by rotenone. Mitochondria, being the major target of rotenone, the effects of melatonin were investigated at the mitochondrial level. Melatonin was able to increase the electron transport chain activity thus preventing the respiratory inhibition by rotenone. The pineal hormone also counteracted the action of rotenone on complex I enzyme. These results suggest melatonin’s ability to potentially limit the free radical generation and thereby modulate the mitochondrial functions. The detection and measurement of lipid peroxidation is the evidence most frequently cited to support the involvement of free radical reactions in toxicology and in human disease. Melatonin also offered significant protection in vivo and in vitro against rotenone induced lipid peroxidation. Since iron plays a major role in oxidative damage and in the progression of Parkinson’s disease, the effect of melatonin on both rotenone and iron induced lipid peroxidation was investigated, the results of which show that melatonin affords protection and this was suggested to be due to its interaction with the rotenone-iron complex that might have formed. Electrochemical studies were further used to characterise the interactions between melatonin, rotenone and iron (III). Melatonin was shown to bind with iron and thus reducing their toxicity. Histological studies were undertaken to assess the effects of melatonin on rotenone induced toxicity on the dopaminergic neurons in the rat brain. Rotenone treated brains showed extensive neuronal damage whereas with melatonin less damage was observed. Rotenone induces apoptosis via reactive oxygen species production and apoptotic cell death has been identified in PD brains. Furthermore, the apoptotic cell death was detected and quantified by the TUNEL staining. Rotenone treated sections showed signs of apoptosis whereas with melatonin, less apoptotic damage was observed. The findings of this study indicate that the neurohormone, melatonin may protect against rotenone-induced neurodegeneration. Since melatonin production falls substantially during ageing, the loss of this antioxidant is theorized to be instrumental in the degenerative processes associated with advanced age. Considering how devastating diseases such as Parkinson’s disease, are to a patient and the patient’s families, the discovery of protective agents are a matter of urgency. Further investigations using the pesticide model will help to determine the involvement of environmental exposure in the pathogenesis of human diseases as well as to test therapeutic strategies for the treatment of such diseases.
- Full Text:
- Date Issued: 2004
Optimization of an intranasal levodopa nanocrystalline formulation for delivery to the brain
- Authors: Kakono, Chiedza
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/420709 , vital:71771
- Description: Thesis embargoed. Probable release date in 2025. , Thesis (MSc (Pharm)) -- Faculty of Pharmacy, Pharmacy, 2023
- Full Text:
- Date Issued: 2023-03-29
- Authors: Kakono, Chiedza
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/420709 , vital:71771
- Description: Thesis embargoed. Probable release date in 2025. , Thesis (MSc (Pharm)) -- Faculty of Pharmacy, Pharmacy, 2023
- Full Text:
- Date Issued: 2023-03-29
Interactions between steroidal anti-inflammatory agents and collagen
- Authors: Kanfer, Isadore
- Date: 1975
- Subjects: Anti-inflammatory agents Collagen
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3849 , http://hdl.handle.net/10962/d1012620
- Description: Much research has been done on the formation of fibrils from solutions of soluble collagen in vitro in order to gain some knowledge of the mechanisms which may occur in vivo. The in vitro formation of fibres from solutions of collagen has been shown to be extremely sensitive to the nature of the solution environment and the presence of added chemical compounds, and thus constitutes an interesting system for the study of collagen-small molecule inter actions. The present study is concerned with the effects of various corticosteroid drugs, used medicinally as anti-inflammatory agents, on collagen in solution. As these corticosteroids are administered to reduce inflammation in conditions such as rheumatoid arthritis and a host of other pathological conditions in which collagen is implicated, this work has been undertaken in order to establish and charac teri ze any binding mechanisms which may be involved. Furthermore, the corticosteroid drugs available commercially in pure form as the free base or as the water-soluble ester salts offer an interesting range of structural and stereochemical variants for the study of their reaction with a complex and biologically important protein molecule such as collagen. A great deal of research on drug- protein interactions (Goldstein, 1949; Meyer and Guttman, 1968a) and more specifically, steroid-protein interactions have been reported over the years (Daughaday, 1959; Sandberg et al., 1966; Villee and Engel, 1961; Westphal , 1971). Comprehensive reports, however, on steroid-collagen interactions in vitro are conspicuously absent from modern scientific literature although relatively superficial accounts have been published (Menczel and Maibach, 1972; Eik-Nes et al., 1954). Although work involving the above has appeared relating specifically to the effects of steroids on collagen biosynthesis both in vivo and in vitro there have been minimal accounts of steroid-collagen interactions tailored to characterize the binding at the molecular level. The effect of corticosteroids on the metabolism of connective tissue has also received special attention (Asboe-Hansen, 1959; Kivirikko, 1953; Nakagawa and Tsurufuji, 1972). Recently, Uitto et al. (1972) reported the effects of several anti-inflammatory corticosteroids on collagen biosynthesis in vitro, whilst Aalto and Kulonen (1972) reported the effects of several antirheumatic drugs on the synthesis of collagen and other proteins in vitro. The interactions between collagen and certain drugs has also been briefly reviewed (Chvapil, 1967). Much data also exists on the binding of a wide range of small molecules and ions with serum albumin (Steinhardt and Reynolds, 1969; Scatchard, 1949; Klotz, 1950). Serum albumin, being specialized for a very general transport function and apparently designed for the purpose of combining with a large range of small molecules, has a proportion of possible reactive sites 'buried' within the molecule itself because of its folded conformation. In addition, serum albumin shows a high degree of cooperative binding in contrast to collagen. The latter molecule, with its larger molecular size and weight is specialized for a biologically structural function and has a higher proportion of possible reactive sites which appear relatively more accessible to ligands. A study of the interactions between corticosteroids and collagen thus provides the opportunity to investigate a protein which is very different from the much studied serum albumin. Because of the limited information available regarding the interaction of steroid drugs and collagen at the molecular level, studies of this nature are relevant to the understanding of the mode of action of steroid compounds which are such an important group of therapeutic substances used in modern medicine.
- Full Text:
- Date Issued: 1975
- Authors: Kanfer, Isadore
- Date: 1975
- Subjects: Anti-inflammatory agents Collagen
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3849 , http://hdl.handle.net/10962/d1012620
- Description: Much research has been done on the formation of fibrils from solutions of soluble collagen in vitro in order to gain some knowledge of the mechanisms which may occur in vivo. The in vitro formation of fibres from solutions of collagen has been shown to be extremely sensitive to the nature of the solution environment and the presence of added chemical compounds, and thus constitutes an interesting system for the study of collagen-small molecule inter actions. The present study is concerned with the effects of various corticosteroid drugs, used medicinally as anti-inflammatory agents, on collagen in solution. As these corticosteroids are administered to reduce inflammation in conditions such as rheumatoid arthritis and a host of other pathological conditions in which collagen is implicated, this work has been undertaken in order to establish and charac teri ze any binding mechanisms which may be involved. Furthermore, the corticosteroid drugs available commercially in pure form as the free base or as the water-soluble ester salts offer an interesting range of structural and stereochemical variants for the study of their reaction with a complex and biologically important protein molecule such as collagen. A great deal of research on drug- protein interactions (Goldstein, 1949; Meyer and Guttman, 1968a) and more specifically, steroid-protein interactions have been reported over the years (Daughaday, 1959; Sandberg et al., 1966; Villee and Engel, 1961; Westphal , 1971). Comprehensive reports, however, on steroid-collagen interactions in vitro are conspicuously absent from modern scientific literature although relatively superficial accounts have been published (Menczel and Maibach, 1972; Eik-Nes et al., 1954). Although work involving the above has appeared relating specifically to the effects of steroids on collagen biosynthesis both in vivo and in vitro there have been minimal accounts of steroid-collagen interactions tailored to characterize the binding at the molecular level. The effect of corticosteroids on the metabolism of connective tissue has also received special attention (Asboe-Hansen, 1959; Kivirikko, 1953; Nakagawa and Tsurufuji, 1972). Recently, Uitto et al. (1972) reported the effects of several anti-inflammatory corticosteroids on collagen biosynthesis in vitro, whilst Aalto and Kulonen (1972) reported the effects of several antirheumatic drugs on the synthesis of collagen and other proteins in vitro. The interactions between collagen and certain drugs has also been briefly reviewed (Chvapil, 1967). Much data also exists on the binding of a wide range of small molecules and ions with serum albumin (Steinhardt and Reynolds, 1969; Scatchard, 1949; Klotz, 1950). Serum albumin, being specialized for a very general transport function and apparently designed for the purpose of combining with a large range of small molecules, has a proportion of possible reactive sites 'buried' within the molecule itself because of its folded conformation. In addition, serum albumin shows a high degree of cooperative binding in contrast to collagen. The latter molecule, with its larger molecular size and weight is specialized for a biologically structural function and has a higher proportion of possible reactive sites which appear relatively more accessible to ligands. A study of the interactions between corticosteroids and collagen thus provides the opportunity to investigate a protein which is very different from the much studied serum albumin. Because of the limited information available regarding the interaction of steroid drugs and collagen at the molecular level, studies of this nature are relevant to the understanding of the mode of action of steroid compounds which are such an important group of therapeutic substances used in modern medicine.
- Full Text:
- Date Issued: 1975
Formulation development, manufacture and evaluation of hydralazine hydrochloride microspheres
- Kangausaru, Shakemore Tinashe
- Authors: Kangausaru, Shakemore Tinashe
- Date: 2017
- Subjects: Hydralazine , Microspheres , Drugs Controlled release , Drugs Design , Drug development , Hypertension Chemotherapy
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/59220 , vital:27482
- Description: Despite improvements in its detection and treatment since the 1970s, hypertension is the most common and important risk factor for cardiovascular diseases. Hypertension is a chronic condition often underdiagnosed and/or inadequately treated in Sub-Saharan Africa. Recent survey results illustrate that the condition continues to contribute significantly to mortality and morbidity in adults and that it is poorly controlled in clinical practice. Hydralazine (HYD) is used either alone or in combination for the management of chronic hypertension, chronic cardiac failure and hypertensive crises. Due to its short plasma half-life of between 2 to 4 hours, HYD is normally administered two to four times daily, therefore making it a potential candidate for inclusion in sustained release formulations. The formulation of sustained release microsphere dosage forms may be useful to improve patient adherence and to achieve predictable and optimised therapeutic plasma concentrations. A stability indicating reversed-phase high performance liquid chromatography (RP-HPLC) method for the quantitation of HYD in pharmaceutical dosage forms was developed and optimised using a Central Composite Design (CCD) approach. UV/Vis detection method was selected as HYD contains an ultraviolet light-absorbing chromophore. The method was validated with respect to linearity and range, limits of quantitation (LOQ) and detection (LOD), accuracy, precision, sensitivity, selectivity and specificity as per International Conference on Harmonisation (ICH) guidelines. The method was applied to commercially available HYD tablets. No interfering peaks were observed from excipients used in the commercially available tablets. Preformulation studies were conducted to ensure the manufacture of high quality, stable sustained release HYD microspheres. The results revealed that there was an interaction between HYD and Carbopol® 971P, therefore Carbopol® polymers were not included during formulation studies. HYD was found to be compatible with Methocel® K100LV, Eudragit® RS PO and Avicel® 101 and HYD formulations were developed and optimised using these excipients. An oil-in-oil (o/o) solvent evaporation technique was selected for the manufacture of HYD microspheres due to its simplicity and to avoid exposure of HYD to moisture that could have been encountered if a water-in-oil (w/o) manufacturing process was used. The selection of o/o solvent evaporation technique was also based on the hydrophilicity of HYD and the polymers selected. Different grades of Methocel® and Eudragit® were selected to evaluate their effect on encapsulation efficiency (EE), in vitro release and microparticle shape and morphology. The best combination of these polymers which resulted in the desired EE, in vitro release, microparticle shape and size were then selected for formulation optimisation. A numerical optimisation approach was used to predict a formulation composition that would produce minimal HYD release initially and maximum HYD release after 12 hours of dissolution testing. The release kinetics of HYD from the manufactured microspheres were established by fitting in vitro release data to several mathematical models. The in vitro release data for the optimised formulations was best described using Higuchi model. The short-term stability of the optimised formulations was established by undertaking stability studies at 4°C, 25 °C/60 % RH and 40 °C/75 % RH. The results revealed that there was no significant change in appearance and physicochemical properties of the microspheres over a period of one month. However, long-term stability studies would be required to determine the shelf-life of the formulations. In addition, a gas chromatographic (GC) method was selected for determining residual amounts of acetone and n-hexane in the optimised formulations. GC methods were developed and optimised by evaluation of process parameters. System suitability testing was performed with respect to resolution, theoretical number of plates and selectivity. Method validation was performed with respect to linearity, range, inter- and intra-day precision, retention time (Rt) precision, limit of quantitation (LOQ) and detection (LOD). A solvent extraction method was used to analyse residual solvents in the optimised formulations. The drying process was sufficient in evaporating acetone and n-hexane from the optimised formulations. Solvent evaporation technique has been successfully used in the manufacture of HYD microspheres. The microspheres have potential for further development, scale up formulation studies and long-term stability studies. , Thesis (MSc) -- Faculty of Pharmacy, Pharmacy, 2017
- Full Text:
- Date Issued: 2017
- Authors: Kangausaru, Shakemore Tinashe
- Date: 2017
- Subjects: Hydralazine , Microspheres , Drugs Controlled release , Drugs Design , Drug development , Hypertension Chemotherapy
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/59220 , vital:27482
- Description: Despite improvements in its detection and treatment since the 1970s, hypertension is the most common and important risk factor for cardiovascular diseases. Hypertension is a chronic condition often underdiagnosed and/or inadequately treated in Sub-Saharan Africa. Recent survey results illustrate that the condition continues to contribute significantly to mortality and morbidity in adults and that it is poorly controlled in clinical practice. Hydralazine (HYD) is used either alone or in combination for the management of chronic hypertension, chronic cardiac failure and hypertensive crises. Due to its short plasma half-life of between 2 to 4 hours, HYD is normally administered two to four times daily, therefore making it a potential candidate for inclusion in sustained release formulations. The formulation of sustained release microsphere dosage forms may be useful to improve patient adherence and to achieve predictable and optimised therapeutic plasma concentrations. A stability indicating reversed-phase high performance liquid chromatography (RP-HPLC) method for the quantitation of HYD in pharmaceutical dosage forms was developed and optimised using a Central Composite Design (CCD) approach. UV/Vis detection method was selected as HYD contains an ultraviolet light-absorbing chromophore. The method was validated with respect to linearity and range, limits of quantitation (LOQ) and detection (LOD), accuracy, precision, sensitivity, selectivity and specificity as per International Conference on Harmonisation (ICH) guidelines. The method was applied to commercially available HYD tablets. No interfering peaks were observed from excipients used in the commercially available tablets. Preformulation studies were conducted to ensure the manufacture of high quality, stable sustained release HYD microspheres. The results revealed that there was an interaction between HYD and Carbopol® 971P, therefore Carbopol® polymers were not included during formulation studies. HYD was found to be compatible with Methocel® K100LV, Eudragit® RS PO and Avicel® 101 and HYD formulations were developed and optimised using these excipients. An oil-in-oil (o/o) solvent evaporation technique was selected for the manufacture of HYD microspheres due to its simplicity and to avoid exposure of HYD to moisture that could have been encountered if a water-in-oil (w/o) manufacturing process was used. The selection of o/o solvent evaporation technique was also based on the hydrophilicity of HYD and the polymers selected. Different grades of Methocel® and Eudragit® were selected to evaluate their effect on encapsulation efficiency (EE), in vitro release and microparticle shape and morphology. The best combination of these polymers which resulted in the desired EE, in vitro release, microparticle shape and size were then selected for formulation optimisation. A numerical optimisation approach was used to predict a formulation composition that would produce minimal HYD release initially and maximum HYD release after 12 hours of dissolution testing. The release kinetics of HYD from the manufactured microspheres were established by fitting in vitro release data to several mathematical models. The in vitro release data for the optimised formulations was best described using Higuchi model. The short-term stability of the optimised formulations was established by undertaking stability studies at 4°C, 25 °C/60 % RH and 40 °C/75 % RH. The results revealed that there was no significant change in appearance and physicochemical properties of the microspheres over a period of one month. However, long-term stability studies would be required to determine the shelf-life of the formulations. In addition, a gas chromatographic (GC) method was selected for determining residual amounts of acetone and n-hexane in the optimised formulations. GC methods were developed and optimised by evaluation of process parameters. System suitability testing was performed with respect to resolution, theoretical number of plates and selectivity. Method validation was performed with respect to linearity, range, inter- and intra-day precision, retention time (Rt) precision, limit of quantitation (LOQ) and detection (LOD). A solvent extraction method was used to analyse residual solvents in the optimised formulations. The drying process was sufficient in evaporating acetone and n-hexane from the optimised formulations. Solvent evaporation technique has been successfully used in the manufacture of HYD microspheres. The microspheres have potential for further development, scale up formulation studies and long-term stability studies. , Thesis (MSc) -- Faculty of Pharmacy, Pharmacy, 2017
- Full Text:
- Date Issued: 2017
The participatory development and implementation of a facilitator’s manual for the promotion of exclusive breastfeeding
- Authors: Katsinde, Shingirai Miranda
- Date: 2016
- Language: English
- Type: Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/4201 , vital:20632
- Description: Background: Breastfeeding is a common practice, but exclusive breastfeeding for the first six months is no longer a cultural norm in the majority of South African communities. Identification of facilitating and constraining factors which affect breastfeeding and exclusive breastfeeding practices is thus important. The promotion of exclusive breastfeeding is essential for improved infant health and development, especially if it takes into consideration the broader cultural and socio-economic aspects influencing these practices. This study follows up on an initial project conducted in Glenmore and Ndwayana, two rural communities in the Eastern Cape, South Africa. The current study aimed at working with community care workers who are associated with two community based organisations, who work within three communities, Glenmore, Ndwayana and Grahamstown. The study objectives were to identify the factors that influence the adoption of exclusive breastfeeding in the communities researched, to conduct workshops with the community workers on exclusive breastfeeding practices, as well as to develop and implement a facilitator’s manual for the promotion of exclusive breastfeeding. Method: Using the community based participatory research approach and the PEN-3 theoretical framework to guide the research process, individual semi-structured interviews were conducted with 14 community care workers to identify the factors that affect breastfeeding and exclusive breastfeeding. Through participatory involvement and a cyclical research process, a facilitator’s manual on breastfeeding was developed, which was used during the workshops in the training of community care workers on breastfeeding and exclusive breastfeeding practices. A pre and post intervention knowledge questionnaire was given to the community care workers to complete before and after the workshops. The facilitator’s manual was modified based on 14 months of consistent interaction with the community care workers, who provided feedback on improving the content as well as on how to enhance the cultural appropriateness of the facilitator’s manual during guided focus group discussions. Readability testing also guided further modification of the facilitator’s manual. Results:The major findings confirmed that exclusive breastfeeding for six months was no longer a common practice in these three communities. Factors affecting breastfeeding and exclusive breastfeeding were classified as perceptions (knowledge attitudes and beliefs), enablers (resources and facilities) and nurturers (people), in accordance with the PEN-3 model thematic categories. The knowledge questionnaire, semi-structured interview results, and data from the workshops showed that the community care workers were not sufficiently equipped with information on breastfeeding and exclusive breastfeeding. The facilitator’s manual development and modification was made possible by inputs from the community care workers and the community based organisation liaisons. A culturally appropriate, community specific facilitator’s manual for the promotion of breastfeeding and exclusive breastfeeding was produced. Conclusion: The factors affecting breastfeeding and exclusive breastfeeding were identified. These factors were useful in facilitating discussions on how to improve breastfeeding and exclusive breastfeeding practices in the communities researched. The facilitator’s manual and the workshops were useful in equipping community care workers with knowledge on breastfeeding and exclusive breastfeeding. The involvement of community based organisations will assist to ensure sustainability of breastfeeding promotion by community care workers by adopting the facilitator’s manual as part of their women and child development programmes.
- Full Text:
- Date Issued: 2016
- Authors: Katsinde, Shingirai Miranda
- Date: 2016
- Language: English
- Type: Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/4201 , vital:20632
- Description: Background: Breastfeeding is a common practice, but exclusive breastfeeding for the first six months is no longer a cultural norm in the majority of South African communities. Identification of facilitating and constraining factors which affect breastfeeding and exclusive breastfeeding practices is thus important. The promotion of exclusive breastfeeding is essential for improved infant health and development, especially if it takes into consideration the broader cultural and socio-economic aspects influencing these practices. This study follows up on an initial project conducted in Glenmore and Ndwayana, two rural communities in the Eastern Cape, South Africa. The current study aimed at working with community care workers who are associated with two community based organisations, who work within three communities, Glenmore, Ndwayana and Grahamstown. The study objectives were to identify the factors that influence the adoption of exclusive breastfeeding in the communities researched, to conduct workshops with the community workers on exclusive breastfeeding practices, as well as to develop and implement a facilitator’s manual for the promotion of exclusive breastfeeding. Method: Using the community based participatory research approach and the PEN-3 theoretical framework to guide the research process, individual semi-structured interviews were conducted with 14 community care workers to identify the factors that affect breastfeeding and exclusive breastfeeding. Through participatory involvement and a cyclical research process, a facilitator’s manual on breastfeeding was developed, which was used during the workshops in the training of community care workers on breastfeeding and exclusive breastfeeding practices. A pre and post intervention knowledge questionnaire was given to the community care workers to complete before and after the workshops. The facilitator’s manual was modified based on 14 months of consistent interaction with the community care workers, who provided feedback on improving the content as well as on how to enhance the cultural appropriateness of the facilitator’s manual during guided focus group discussions. Readability testing also guided further modification of the facilitator’s manual. Results:The major findings confirmed that exclusive breastfeeding for six months was no longer a common practice in these three communities. Factors affecting breastfeeding and exclusive breastfeeding were classified as perceptions (knowledge attitudes and beliefs), enablers (resources and facilities) and nurturers (people), in accordance with the PEN-3 model thematic categories. The knowledge questionnaire, semi-structured interview results, and data from the workshops showed that the community care workers were not sufficiently equipped with information on breastfeeding and exclusive breastfeeding. The facilitator’s manual development and modification was made possible by inputs from the community care workers and the community based organisation liaisons. A culturally appropriate, community specific facilitator’s manual for the promotion of breastfeeding and exclusive breastfeeding was produced. Conclusion: The factors affecting breastfeeding and exclusive breastfeeding were identified. These factors were useful in facilitating discussions on how to improve breastfeeding and exclusive breastfeeding practices in the communities researched. The facilitator’s manual and the workshops were useful in equipping community care workers with knowledge on breastfeeding and exclusive breastfeeding. The involvement of community based organisations will assist to ensure sustainability of breastfeeding promotion by community care workers by adopting the facilitator’s manual as part of their women and child development programmes.
- Full Text:
- Date Issued: 2016
Formulation and evaluation of captopril loaded polymethacrylate and hydroxypropyl methycellulose microcapsules
- Khamanga, Sandile Maswazi Malungelo
- Authors: Khamanga, Sandile Maswazi Malungelo
- Date: 2010
- Subjects: Hypertension -- Treatment , Hypertension -- Chemotherapy , Angiotensin converting enzyme -- Inhibitors , Hypotensive agents -- Development , Pharmacokinetics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3860 , http://hdl.handle.net/10962/d1013443
- Description: Angiotensin-converting enzyme (ACE) inhibitors are some of the most commonly prescribed medications for hypertension. They are cited in many papers as the treatment most often recommended by guidelines and favoured over other antihypertensive drugs as first-line agents especially when other high-risk conditions are present, such as diabetic nephropathy. The development of captopril (CPT) was amongst the earliest successes of the revolutionary concept of structure-based drug design. Due to its relatively poor pharmacokinetic profile or short half-life of about 1 hour, the formulation of sustained-release microcapsule dosage form is useful to improve patient compliance and to achieve predictable and optimized therapeutic plasma concentrations. Currently, CPT is mainly administered in tablet form. One of the difficulties of CPT formulation has been reported to be its instability in aqueous solutions. CPT is characterized by a lack of a strong chromophore and, therefore, not able to absorb at the more useful UV–Vis region of the spectrum. For this reason, an accurate, simple, reproducible, and sensitive HPLC-ECD method was developed and validated for the determination of CPT in dosage forms. The method was successfully applied for the determination of CPT in commercial and developed formulations. Possible drug-excipient and excipient-excipient interactions were investigated prior to formulating CPT microcapsules because successful formulation of a stable and effective solid dosage form depends on careful selection of excipients. Nuclear magnetic resonance spectroscopy, Fourier transform infra-red spectroscopy (FT-IR), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) were used for the identification and purity testing of CPT and excipients. The studies revealed no thermal changes during stress testing of binary and whole mixtures which indicate absence of solid state interactions. There were no shifts, appearance and disappearance in the endothermic or exothermic peaks and on the change of other associated enthalpy values on thermal curves obtained with DSC method. Characteristic peaks for common functional groups in the FT-IR were present in all the mixtures indicating the absence of incompatibility. The techniques used in this study can be said to have been efficient in the characterization and evaluation of the drug and excipients. The technique of microencapsulation by oil-in-oil was used to prepare CPT microcapsules. The effects of polymer molecular weight, homogenizing speed on the particle size, flow properties, morphology, surface properties and release characteristics of the prepared CPT microcapsules were examined. In order to decrease the complexity of the analysis and reduce cost response surface methodology using best polynomial equations was successfully used to quantify the effect of the formulation variables and develop an optimized formulation thereby minimizing the number of experimental trials. There was a burst effect during the first stage of dissolution. Scanning electron microscopy (SEM) results indicated that the initial burst effect observed in drug release could be attributed to dissolution of CPT crystals present at the surface or embedded in the superficial layer of the matrix. During the preparation of microcapsules, the drug might have been trapped near the surface of the microcapsules and or might have diffused quickly through the porous surface. The release kinetics of CPT from most formulations followed Fickian diffusion mechanism. SEM photographs showed that diffusion took place through pores at the surface of the microcapsules. The Kopcha model diffusion and erosion terms showed predominance of diffusion relative to swelling or erosion throughout the entire test period. Drug release mechanism was also confirmed by Makoid-Banakar and Korsmeyer-Peppas models exponents which further support diffusion release mechanism in most formulations. The models postulate that the total of drug release is a summation of a couple of mechanisms; burst release, relaxation induced controlled-release and diffusional release. Inspection of the 2D contour and 3D response surfaces allowed the determination of the geometrical nature of the surfaces and further providing results about the interaction of the different variables used in central composite design (CCD). The wide variation indicated that the factor combinations resulted in different drug release rates. Lagrange, canonical and mathematical modelling were used to determine the nature of the stationery point of the models. This represented the optimal variables or stationery points where there is interaction in the experimental space. It is difficult to understand the shape of a fitted response by mere inspection of the algebraic polynomial when there are many independent variables in the model. Canonical and Lagrange analyses facilitated the interpretation of the surface plots after a mathematical transformation of the original variables into new variables. In conclusion, these results suggest the potential application of Eudragit® / Methocel® microcapsules as suitable sustained-release drug delivery system for CPT.
- Full Text:
- Date Issued: 2010
- Authors: Khamanga, Sandile Maswazi Malungelo
- Date: 2010
- Subjects: Hypertension -- Treatment , Hypertension -- Chemotherapy , Angiotensin converting enzyme -- Inhibitors , Hypotensive agents -- Development , Pharmacokinetics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3860 , http://hdl.handle.net/10962/d1013443
- Description: Angiotensin-converting enzyme (ACE) inhibitors are some of the most commonly prescribed medications for hypertension. They are cited in many papers as the treatment most often recommended by guidelines and favoured over other antihypertensive drugs as first-line agents especially when other high-risk conditions are present, such as diabetic nephropathy. The development of captopril (CPT) was amongst the earliest successes of the revolutionary concept of structure-based drug design. Due to its relatively poor pharmacokinetic profile or short half-life of about 1 hour, the formulation of sustained-release microcapsule dosage form is useful to improve patient compliance and to achieve predictable and optimized therapeutic plasma concentrations. Currently, CPT is mainly administered in tablet form. One of the difficulties of CPT formulation has been reported to be its instability in aqueous solutions. CPT is characterized by a lack of a strong chromophore and, therefore, not able to absorb at the more useful UV–Vis region of the spectrum. For this reason, an accurate, simple, reproducible, and sensitive HPLC-ECD method was developed and validated for the determination of CPT in dosage forms. The method was successfully applied for the determination of CPT in commercial and developed formulations. Possible drug-excipient and excipient-excipient interactions were investigated prior to formulating CPT microcapsules because successful formulation of a stable and effective solid dosage form depends on careful selection of excipients. Nuclear magnetic resonance spectroscopy, Fourier transform infra-red spectroscopy (FT-IR), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) were used for the identification and purity testing of CPT and excipients. The studies revealed no thermal changes during stress testing of binary and whole mixtures which indicate absence of solid state interactions. There were no shifts, appearance and disappearance in the endothermic or exothermic peaks and on the change of other associated enthalpy values on thermal curves obtained with DSC method. Characteristic peaks for common functional groups in the FT-IR were present in all the mixtures indicating the absence of incompatibility. The techniques used in this study can be said to have been efficient in the characterization and evaluation of the drug and excipients. The technique of microencapsulation by oil-in-oil was used to prepare CPT microcapsules. The effects of polymer molecular weight, homogenizing speed on the particle size, flow properties, morphology, surface properties and release characteristics of the prepared CPT microcapsules were examined. In order to decrease the complexity of the analysis and reduce cost response surface methodology using best polynomial equations was successfully used to quantify the effect of the formulation variables and develop an optimized formulation thereby minimizing the number of experimental trials. There was a burst effect during the first stage of dissolution. Scanning electron microscopy (SEM) results indicated that the initial burst effect observed in drug release could be attributed to dissolution of CPT crystals present at the surface or embedded in the superficial layer of the matrix. During the preparation of microcapsules, the drug might have been trapped near the surface of the microcapsules and or might have diffused quickly through the porous surface. The release kinetics of CPT from most formulations followed Fickian diffusion mechanism. SEM photographs showed that diffusion took place through pores at the surface of the microcapsules. The Kopcha model diffusion and erosion terms showed predominance of diffusion relative to swelling or erosion throughout the entire test period. Drug release mechanism was also confirmed by Makoid-Banakar and Korsmeyer-Peppas models exponents which further support diffusion release mechanism in most formulations. The models postulate that the total of drug release is a summation of a couple of mechanisms; burst release, relaxation induced controlled-release and diffusional release. Inspection of the 2D contour and 3D response surfaces allowed the determination of the geometrical nature of the surfaces and further providing results about the interaction of the different variables used in central composite design (CCD). The wide variation indicated that the factor combinations resulted in different drug release rates. Lagrange, canonical and mathematical modelling were used to determine the nature of the stationery point of the models. This represented the optimal variables or stationery points where there is interaction in the experimental space. It is difficult to understand the shape of a fitted response by mere inspection of the algebraic polynomial when there are many independent variables in the model. Canonical and Lagrange analyses facilitated the interpretation of the surface plots after a mathematical transformation of the original variables into new variables. In conclusion, these results suggest the potential application of Eudragit® / Methocel® microcapsules as suitable sustained-release drug delivery system for CPT.
- Full Text:
- Date Issued: 2010
Formulation and assessment of verapamil sustained release tablets
- Khamanga, Sandile Maswazi Malungelo
- Authors: Khamanga, Sandile Maswazi Malungelo
- Date: 2005
- Subjects: Verapamil , Tablets (Medicine) , Drugs -- Administration , Cardiovascular agents , Calcium -- Antagonists , Drugs -- Controlled release
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3869 , http://hdl.handle.net/10962/d1018236
- Description: The oral route of drug administration is most extensively used due to the obvious ease of administration. Verapamil hydrochloride is a WHO listed phenylalkylarnine, L-type calcium channel antagonist that is mainly indicated for cardiovascular disorders such as angina pectoris, supraventricular tachycardia and hypertension. Due to its relatively short half-life of approximately 4.0 hours, the formulation of a sustained-release dosage form is useful to improve patient compliance and to achieve predictable and optimized therapeutic plasma concentrations. Direct compression and wet granulation were initially used as methods for tablet manufacture. The direct compression method of manufacture produced tablets that exhibited formulation and manufacturing difficulties. Mini-tablets containing veraparnil hydrochloride were then prepared by wet granulation using Surelease® E-7-19010.and Eudragit® NE 30D as the granulating agents after which the granules were incorporated with an hydrophilic matrix material, Carbopol® 974P NF. Granule and powder blends were evaluated using the angle of repose, loose and tapped bulk density, Can's compressibility index, Hausner's ratio and drug content. Granules with good flow properties and satisfactory compressibility were used for further studies. Tablets were subjected to thickness, diameter and weight variation tests, crushing strength, tensile strength, friability and content uniformity studies. Tablets that showed acceptable pharmaco-technical properties were selected for further analysis. Drug content uniformity and dissolution release rates were determined using a validated isocratic HPLC method. Initially, USP apparatus 1 and 3 dissolution apparatus were used to determine in-vitro drug release rates from the formulations over a 22-hour period. USP apparatus 3 was finally selected as it offers the advantages of mimicking, in part, the changes in the physicochemical environment experienced by products in the gastro-intestinal tract. Differences in release rates between the test formulations and a commercially available product, Isoptin® SR were observed at different pH's using USP apparatus 1. The release of veraparnil hydrochloride from matrix tablets was pH dependent and was markedly reduced at higher pH values. This may be due, in part, to the poor solubility of veraparnil hydrochloride at these pH values and also the possible interaction of verapamil hydrochloride with anionic polymers used in these formulations. Swelling and erosion behaviour of the tablets were evaluated and differences in behaviour were observed which may be attributed to the physico-chemical characteristics of the polymers used in this study. In-vitro dissolution profiles were characterized by the difference (j1) and similarity factor (j2) and also by a new similarity factor, Sct. In addition, the mechanism of drug release from these dosage forms was mainly evaluated using the Korsmeyer-Peppas model and the kinetics of drug release assessed using other models, including Zero order, First order, Higuchi, HixsonCrowell, Weibull and the Baker-Lonsdale model. Dissolution kinetics were best described by application of the Weibull model, and the Korsmeyer-Peppas model. The release exponent, n, confirmed that drug release from these dosage forms was due to the mixed effects of diffusion and swelling and therefore, anomalous release kinetics are predominant. In conclusion, two test batches were found to be comparable to the reference product Isoptin® SR with respect to their in-vitro release profiles.
- Full Text:
- Date Issued: 2005
- Authors: Khamanga, Sandile Maswazi Malungelo
- Date: 2005
- Subjects: Verapamil , Tablets (Medicine) , Drugs -- Administration , Cardiovascular agents , Calcium -- Antagonists , Drugs -- Controlled release
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3869 , http://hdl.handle.net/10962/d1018236
- Description: The oral route of drug administration is most extensively used due to the obvious ease of administration. Verapamil hydrochloride is a WHO listed phenylalkylarnine, L-type calcium channel antagonist that is mainly indicated for cardiovascular disorders such as angina pectoris, supraventricular tachycardia and hypertension. Due to its relatively short half-life of approximately 4.0 hours, the formulation of a sustained-release dosage form is useful to improve patient compliance and to achieve predictable and optimized therapeutic plasma concentrations. Direct compression and wet granulation were initially used as methods for tablet manufacture. The direct compression method of manufacture produced tablets that exhibited formulation and manufacturing difficulties. Mini-tablets containing veraparnil hydrochloride were then prepared by wet granulation using Surelease® E-7-19010.and Eudragit® NE 30D as the granulating agents after which the granules were incorporated with an hydrophilic matrix material, Carbopol® 974P NF. Granule and powder blends were evaluated using the angle of repose, loose and tapped bulk density, Can's compressibility index, Hausner's ratio and drug content. Granules with good flow properties and satisfactory compressibility were used for further studies. Tablets were subjected to thickness, diameter and weight variation tests, crushing strength, tensile strength, friability and content uniformity studies. Tablets that showed acceptable pharmaco-technical properties were selected for further analysis. Drug content uniformity and dissolution release rates were determined using a validated isocratic HPLC method. Initially, USP apparatus 1 and 3 dissolution apparatus were used to determine in-vitro drug release rates from the formulations over a 22-hour period. USP apparatus 3 was finally selected as it offers the advantages of mimicking, in part, the changes in the physicochemical environment experienced by products in the gastro-intestinal tract. Differences in release rates between the test formulations and a commercially available product, Isoptin® SR were observed at different pH's using USP apparatus 1. The release of veraparnil hydrochloride from matrix tablets was pH dependent and was markedly reduced at higher pH values. This may be due, in part, to the poor solubility of veraparnil hydrochloride at these pH values and also the possible interaction of verapamil hydrochloride with anionic polymers used in these formulations. Swelling and erosion behaviour of the tablets were evaluated and differences in behaviour were observed which may be attributed to the physico-chemical characteristics of the polymers used in this study. In-vitro dissolution profiles were characterized by the difference (j1) and similarity factor (j2) and also by a new similarity factor, Sct. In addition, the mechanism of drug release from these dosage forms was mainly evaluated using the Korsmeyer-Peppas model and the kinetics of drug release assessed using other models, including Zero order, First order, Higuchi, HixsonCrowell, Weibull and the Baker-Lonsdale model. Dissolution kinetics were best described by application of the Weibull model, and the Korsmeyer-Peppas model. The release exponent, n, confirmed that drug release from these dosage forms was due to the mixed effects of diffusion and swelling and therefore, anomalous release kinetics are predominant. In conclusion, two test batches were found to be comparable to the reference product Isoptin® SR with respect to their in-vitro release profiles.
- Full Text:
- Date Issued: 2005
A study of possible interactions between the pineal gland and the opioidergic system
- Authors: Khan, Razeeya B
- Date: 1990
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3729 , http://hdl.handle.net/10962/d1001468
- Description: Recent observations suggest a link between the pineal gland and the opioid system. Possible areas of interaction between the pineal gland and the opioidergic system in Wistar rats were investigated. The effect of opioids on the pineal gland in organ culture was monitored. Neither morphine, methadone nor the opioid antagonist naloxone was found to affect [¹⁴C]-serotonin metabolism by the pineal gland in vitro. Both the pineal gland and the opioid system are influenced by exposure to stressful stimuli. Morphine and melatonin had protective effects on stress-induced gastric lesions. The ability of melatonin to inhibit lesion formation was found not to be exerted via an opioidergic mechanism. Evidence has been obtained for a possible modulation of the stress response by the pineal gland . The opioid drugs are the most potent analgesic agents available. A possible interaction between the opioid system and the pineal gland in the modulation of the response to noxious stimuli was investigated. An intact pineal gland was found to be necessary for the manifestation of the nocturnally increased response of rats to noxious stimuli
- Full Text:
- Date Issued: 1990
- Authors: Khan, Razeeya B
- Date: 1990
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3729 , http://hdl.handle.net/10962/d1001468
- Description: Recent observations suggest a link between the pineal gland and the opioid system. Possible areas of interaction between the pineal gland and the opioidergic system in Wistar rats were investigated. The effect of opioids on the pineal gland in organ culture was monitored. Neither morphine, methadone nor the opioid antagonist naloxone was found to affect [¹⁴C]-serotonin metabolism by the pineal gland in vitro. Both the pineal gland and the opioid system are influenced by exposure to stressful stimuli. Morphine and melatonin had protective effects on stress-induced gastric lesions. The ability of melatonin to inhibit lesion formation was found not to be exerted via an opioidergic mechanism. Evidence has been obtained for a possible modulation of the stress response by the pineal gland . The opioid drugs are the most potent analgesic agents available. A possible interaction between the opioid system and the pineal gland in the modulation of the response to noxious stimuli was investigated. An intact pineal gland was found to be necessary for the manifestation of the nocturnally increased response of rats to noxious stimuli
- Full Text:
- Date Issued: 1990
Formulation and assessment of monolithic beta blocker sustained release tablets prepared by direct compression
- Authors: Kieser, Leith Faye
- Date: 2002
- Subjects: Drugs -- Dosage forms , Drugs -- Administration , Pharmacology, Experimental , Adrenergic beta blockers , Tablets (Medicine) , Tableting , Neuropharmacology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3764 , http://hdl.handle.net/10962/d1003242 , Drugs -- Dosage forms , Drugs -- Administration , Pharmacology, Experimental , Adrenergic beta blockers , Tablets (Medicine) , Tableting , Neuropharmacology
- Description: Beta blockers are commonly prescribed for the chronic treatment of hypertension, one of the most prolific disease states worldwide. The beta blockers selected for this study include acebutolol hydrochloride, labetalol hydrochloride, metoprolol tartrate oxprenolol hydrochloride and propranolol hydrochloride. All of these compounds have a short elimination half-life, necessitating multiple dose per day regimens and therefore the development of sustained release dosage forms incorporating these agents was considered beneficial in terms of extending the dosing interval, with the aim of improving patient compliance and subsequent therapeutic outcomes. Preformulation studies that were conducted included moisture content analysis by Karl Fischer titration, and DSC, a method used to predict potential interactions between the drugs and tablet excipients. Tablets were manufactured by both wet granulation and direct compression techniques, and the resultant drug release characteristics were evaluated using the USP Apparatus 3(BIO.DIS). A validated isocratic HPLC method, capable of separating the five drug candidates simultaneously, was developed and used for the analysis of drug samples. Tablet quality was assessed using analyses that included the physical assessment of weight, diameter, thickness, hardness and friability, as well as content uniformity of tablets, before and after dissolution testing. Direct compression tablet formulations containing each of the five beta blockers were successfully adapted from a prototype wet granulation matrix tablet containing metoprolol tartrate, and various formulation variables were investigated to establish,their effect on the rate and extent of drug release from these tablets. The grade and quantity of ethylcellulose used in the wet granulation and direct compression formulae influenced the release rate of some drug candidates. In addition, an alternative formulation method, involving freeze-drying of the drug with an ethylcellulose dispersion, was shown to have potential for altering release rates further. Anti-frictional agents, talc and colloidal silicon dioxide, did not affect drug release from these matrices,however, they affected the physical character:istics such as tablet weight and thickness, of the resultant tablets. All of the matrix tablets formulated were shown to release drug according to square root of time kinetics, in a sustained manner over a 22 hour period.
- Full Text:
- Date Issued: 2002
- Authors: Kieser, Leith Faye
- Date: 2002
- Subjects: Drugs -- Dosage forms , Drugs -- Administration , Pharmacology, Experimental , Adrenergic beta blockers , Tablets (Medicine) , Tableting , Neuropharmacology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3764 , http://hdl.handle.net/10962/d1003242 , Drugs -- Dosage forms , Drugs -- Administration , Pharmacology, Experimental , Adrenergic beta blockers , Tablets (Medicine) , Tableting , Neuropharmacology
- Description: Beta blockers are commonly prescribed for the chronic treatment of hypertension, one of the most prolific disease states worldwide. The beta blockers selected for this study include acebutolol hydrochloride, labetalol hydrochloride, metoprolol tartrate oxprenolol hydrochloride and propranolol hydrochloride. All of these compounds have a short elimination half-life, necessitating multiple dose per day regimens and therefore the development of sustained release dosage forms incorporating these agents was considered beneficial in terms of extending the dosing interval, with the aim of improving patient compliance and subsequent therapeutic outcomes. Preformulation studies that were conducted included moisture content analysis by Karl Fischer titration, and DSC, a method used to predict potential interactions between the drugs and tablet excipients. Tablets were manufactured by both wet granulation and direct compression techniques, and the resultant drug release characteristics were evaluated using the USP Apparatus 3(BIO.DIS). A validated isocratic HPLC method, capable of separating the five drug candidates simultaneously, was developed and used for the analysis of drug samples. Tablet quality was assessed using analyses that included the physical assessment of weight, diameter, thickness, hardness and friability, as well as content uniformity of tablets, before and after dissolution testing. Direct compression tablet formulations containing each of the five beta blockers were successfully adapted from a prototype wet granulation matrix tablet containing metoprolol tartrate, and various formulation variables were investigated to establish,their effect on the rate and extent of drug release from these tablets. The grade and quantity of ethylcellulose used in the wet granulation and direct compression formulae influenced the release rate of some drug candidates. In addition, an alternative formulation method, involving freeze-drying of the drug with an ethylcellulose dispersion, was shown to have potential for altering release rates further. Anti-frictional agents, talc and colloidal silicon dioxide, did not affect drug release from these matrices,however, they affected the physical character:istics such as tablet weight and thickness, of the resultant tablets. All of the matrix tablets formulated were shown to release drug according to square root of time kinetics, in a sustained manner over a 22 hour period.
- Full Text:
- Date Issued: 2002
The development and assessment of a fixed dose combination tablet of Ranitidine and Metronidazole
- Authors: King'ori, Loti David
- Date: 2011 , 2011-04-07
- Subjects: Ulcers -- Treatment , Ranitidine -- Evaluation , Metronidazole -- Evaluation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3858 , http://hdl.handle.net/10962/d1013359
- Description: The oral route of drug administration is convenient since it is acceptable to most patients and the manufacturing processes used to produce tablets and capsules are relatively simple when compared to those used to manufacture other types of dosage forms. Metronidazole (MTZ) and Ranitidine (RTD) have been used in combination, as part of triple therapy for the treatment of ulcers. However the use of large numbers of tablets and long duration of therapy makes adherence to drug treatment challenging for patients. Therefore the formulation of a fixed dose combination (FDC) of MTZ and RTD may improve patient adherence to therapy and consequently may reduce morbidity and mortality due to ulcers. A stability indicating HPLC method for the simultaneous analysis of MTZ and RTD was developed and validated according to the International Conference on Harmonization (ICH) guidelines. The method was sensitive, selective, precise, accurate and linear.Preformulation studies were performed on the active pharmaceutical ingredients (API) alone and in combination with potential excipients. Differential scanning calorimetry (DSC) studies revealed a potential interaction between MTZ and RTD, however the interaction was not apparent following IR analysis of the same samples. DSC analyses of the API in combination with potential excipients revealed that the compounds were compatible with most materials with the exception of a binary mixture of RTD and Dibasic calcium phosphate (DCP) that exhibited a potential interaction. Thermal gravimetric analysis (TGA) of MTZ and RTD revealed that both compounds exhibited thermal stability. The Carrs Index (CI) and Hausner Ratio (HR) values of MTZ and RTD indicated that both compounds exhibited poor flow and compressibility properties, whereas the CI and HR values for (Microcrystalline cellulose) MCC and DCP indicated better flowability and compressibility characteristics.Direct compression and wet granulation processes were assessed to identify a suitable method of manufacture of FDC tablets of MTZ and RTD. The blends were evaluated using bulk and tapped density and the resultant tablets were evaluated for weight uniformity, crushing strength, tensile strength and disintegration time. The wet granulation method of manufacture produced tablets that showed acceptable pharmacotechnical properties: this approach was therefore used as the method of manufacture of FDC tablets of MTZ and RTD. Tablet formulations comprised of API, viz. MTZ and RTD and different compositions of MCC, DCP, Sodium starch glycolate (SSG) and Croscarmellose sodium (CCS), were manufactured in order to screen for an appropriate diluent and disintegrant composition for use in response surface studies. Assays of tablet content and in vitro drug release were undertaken using the validated HPLC method. Tablets in which MCC and CCS were used appeared to produce better assay and dissolution results as compared to those manufactured using DCP and SSG. Consequently a formulation comprised of MCC and CCS was selected and used in studies in which the effect(s) of level two formulation and composition changes as described in the Scale and Post Approval Changes for Immediate Release (SUPAC-IR) Guidelines on tablet disintegration and in vitro release were assessed. A Box-Behnken statistical design was used for the investigation of the effect of input factors, viz. CCS, (Polyvinyl pyrollidone K30) PVP-K30 and magnesium stearate on measured responses, viz. disintegration time and percent drug release in 10 minutes (Q10). CCS appeared to have an inverse linear relationship on disintegration time and a linear relationship with the Q10 for MTZ and RTD, whereas PVP-K30 and magnesium stearate appeared to have an antagonistic effect on the measured responses. Furthermore CCS and magnesium stearate exhibited an interaction that had an agonistic effect on the Q10 value for RTD. A numerical optimization approach was used to predict a formulation composition that would produce tablets that exhibited a disintegration time and Q10 values for MTZ and RTD that fell within the constraints set in our laboratory. The resultant model was found to be accurate and had a percent prediction error of < 5% for all measured response variables.FDC tablets of MTZ and RTD have been successfully produced. The disintegration of the tablet and dissolution of the API were within compendial specifications and the tablets are of suitable quality and have the potential to be further investigated to reduce the pill burden in the treatment of ulcers.
- Full Text:
- Date Issued: 2011
- Authors: King'ori, Loti David
- Date: 2011 , 2011-04-07
- Subjects: Ulcers -- Treatment , Ranitidine -- Evaluation , Metronidazole -- Evaluation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3858 , http://hdl.handle.net/10962/d1013359
- Description: The oral route of drug administration is convenient since it is acceptable to most patients and the manufacturing processes used to produce tablets and capsules are relatively simple when compared to those used to manufacture other types of dosage forms. Metronidazole (MTZ) and Ranitidine (RTD) have been used in combination, as part of triple therapy for the treatment of ulcers. However the use of large numbers of tablets and long duration of therapy makes adherence to drug treatment challenging for patients. Therefore the formulation of a fixed dose combination (FDC) of MTZ and RTD may improve patient adherence to therapy and consequently may reduce morbidity and mortality due to ulcers. A stability indicating HPLC method for the simultaneous analysis of MTZ and RTD was developed and validated according to the International Conference on Harmonization (ICH) guidelines. The method was sensitive, selective, precise, accurate and linear.Preformulation studies were performed on the active pharmaceutical ingredients (API) alone and in combination with potential excipients. Differential scanning calorimetry (DSC) studies revealed a potential interaction between MTZ and RTD, however the interaction was not apparent following IR analysis of the same samples. DSC analyses of the API in combination with potential excipients revealed that the compounds were compatible with most materials with the exception of a binary mixture of RTD and Dibasic calcium phosphate (DCP) that exhibited a potential interaction. Thermal gravimetric analysis (TGA) of MTZ and RTD revealed that both compounds exhibited thermal stability. The Carrs Index (CI) and Hausner Ratio (HR) values of MTZ and RTD indicated that both compounds exhibited poor flow and compressibility properties, whereas the CI and HR values for (Microcrystalline cellulose) MCC and DCP indicated better flowability and compressibility characteristics.Direct compression and wet granulation processes were assessed to identify a suitable method of manufacture of FDC tablets of MTZ and RTD. The blends were evaluated using bulk and tapped density and the resultant tablets were evaluated for weight uniformity, crushing strength, tensile strength and disintegration time. The wet granulation method of manufacture produced tablets that showed acceptable pharmacotechnical properties: this approach was therefore used as the method of manufacture of FDC tablets of MTZ and RTD. Tablet formulations comprised of API, viz. MTZ and RTD and different compositions of MCC, DCP, Sodium starch glycolate (SSG) and Croscarmellose sodium (CCS), were manufactured in order to screen for an appropriate diluent and disintegrant composition for use in response surface studies. Assays of tablet content and in vitro drug release were undertaken using the validated HPLC method. Tablets in which MCC and CCS were used appeared to produce better assay and dissolution results as compared to those manufactured using DCP and SSG. Consequently a formulation comprised of MCC and CCS was selected and used in studies in which the effect(s) of level two formulation and composition changes as described in the Scale and Post Approval Changes for Immediate Release (SUPAC-IR) Guidelines on tablet disintegration and in vitro release were assessed. A Box-Behnken statistical design was used for the investigation of the effect of input factors, viz. CCS, (Polyvinyl pyrollidone K30) PVP-K30 and magnesium stearate on measured responses, viz. disintegration time and percent drug release in 10 minutes (Q10). CCS appeared to have an inverse linear relationship on disintegration time and a linear relationship with the Q10 for MTZ and RTD, whereas PVP-K30 and magnesium stearate appeared to have an antagonistic effect on the measured responses. Furthermore CCS and magnesium stearate exhibited an interaction that had an agonistic effect on the Q10 value for RTD. A numerical optimization approach was used to predict a formulation composition that would produce tablets that exhibited a disintegration time and Q10 values for MTZ and RTD that fell within the constraints set in our laboratory. The resultant model was found to be accurate and had a percent prediction error of < 5% for all measured response variables.FDC tablets of MTZ and RTD have been successfully produced. The disintegration of the tablet and dissolution of the API were within compendial specifications and the tablets are of suitable quality and have the potential to be further investigated to reduce the pill burden in the treatment of ulcers.
- Full Text:
- Date Issued: 2011
Isolation, structural characterisation and evaluation of cytotoxic activity of natural products from selected South African marine red algae
- Authors: Knott, Michael George
- Date: 2012
- Subjects: Marine algae -- South Africa , Red algae -- South Africa , Pharmaceutical chemistry
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3862 , http://hdl.handle.net/10962/d1015460
- Description: The medicinal chemistry of selected marine algae indigenous to South Africa was investigated. Following the isolation and characterisation of a number of new and known compounds, the associated in vitro cytotoxic profiles of these new compounds was investigated. Plocamium maxillosum yielded two new cyclic polyhalogenated monoterpenes which were characterised as 2E-chloromethine-4E-chlorovinyl-4-methyl-5-cyclohexen-1-one (2.1) and 2Z-chloromethine-4E-chlorovinyl-4-methyl-5-cyclohexen-1-one (2.2) on the basis of one and two dimensional NMR spectroscopic data and MS analysis. These compounds were also found to have good cytotoxic activity against breast cancer cell lines. Although these compounds are based on a regular monoterpene skeleton, they represent an uncommon feature not often seen in cyclic halogenated monoterpenes from marine algae. Plocamium robertiae yielded one new cyclic polyhalogenated monoterpene identified as 4,5- dibromo-5-chloromethyl-1-chlorovinyl-2-chloro-methylcyclohexane (2.6) and one known compound called 2,4-dichloro-1-chlorovinyl-1-methylcyclohexane-5-ene or Plocamene D (2.9). Portieria hornemannii was collected from Port Edward in Natal and yielded three new compounds, namely; 3Z-1,6-dibromo-3-(bromomethylidene)-2,7-dichloro-7-methyloctane (3.1), 1E,3Z-1,6-dibromo-3-(bromomethylidene)-7-chloro-7-methyloct-1-ene (3.2), 1Z,3Z- 1,6-dibromo-3-(bromomethylidene)-7-chloro-7-methyloct-1-ene (3.3), and one known compound, namely; 3S,6R-6-bromo-3-(bromomethyl)-3,7-dichloro-7-methyloct-1-ene (3.4). Compounds 3.1 and 3.2 showed no cytotoxic activity against breast cancer cells. Another Portieria hornemannii sample was collected from Noordhoek in the Eastern Cape, it yielded one known compound referred to as 3Z-6-bromo-3-(bromomethylidene)-2,7- dichloro-7-methyloct-1-ene (3.5), as well as one new compound called portieric acid A (3.6) or 5-bromo-2-(bromomethylidene)-6-chloro-6-methylheptanoic acid. Portieric acid A showed slight cytotoxic activity and also represents a new class of compound within the genus Portieria. The isolation of secondary metabolites from the South African red alga, Laurencia glomerata, yielded two known compounds; 7-hydroxylaurene (4.9) and cis-neolaurencenyne (4.12), as well as one chamigrane related compound (4.11). Laurencia flexuosa yielded one known compound called 3Z-bromofucin (4.13). Using 1H NMR, GC and molecular systematics, a novel method for identifying different species of Laurencia was also investigated.
- Full Text:
- Date Issued: 2012
- Authors: Knott, Michael George
- Date: 2012
- Subjects: Marine algae -- South Africa , Red algae -- South Africa , Pharmaceutical chemistry
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3862 , http://hdl.handle.net/10962/d1015460
- Description: The medicinal chemistry of selected marine algae indigenous to South Africa was investigated. Following the isolation and characterisation of a number of new and known compounds, the associated in vitro cytotoxic profiles of these new compounds was investigated. Plocamium maxillosum yielded two new cyclic polyhalogenated monoterpenes which were characterised as 2E-chloromethine-4E-chlorovinyl-4-methyl-5-cyclohexen-1-one (2.1) and 2Z-chloromethine-4E-chlorovinyl-4-methyl-5-cyclohexen-1-one (2.2) on the basis of one and two dimensional NMR spectroscopic data and MS analysis. These compounds were also found to have good cytotoxic activity against breast cancer cell lines. Although these compounds are based on a regular monoterpene skeleton, they represent an uncommon feature not often seen in cyclic halogenated monoterpenes from marine algae. Plocamium robertiae yielded one new cyclic polyhalogenated monoterpene identified as 4,5- dibromo-5-chloromethyl-1-chlorovinyl-2-chloro-methylcyclohexane (2.6) and one known compound called 2,4-dichloro-1-chlorovinyl-1-methylcyclohexane-5-ene or Plocamene D (2.9). Portieria hornemannii was collected from Port Edward in Natal and yielded three new compounds, namely; 3Z-1,6-dibromo-3-(bromomethylidene)-2,7-dichloro-7-methyloctane (3.1), 1E,3Z-1,6-dibromo-3-(bromomethylidene)-7-chloro-7-methyloct-1-ene (3.2), 1Z,3Z- 1,6-dibromo-3-(bromomethylidene)-7-chloro-7-methyloct-1-ene (3.3), and one known compound, namely; 3S,6R-6-bromo-3-(bromomethyl)-3,7-dichloro-7-methyloct-1-ene (3.4). Compounds 3.1 and 3.2 showed no cytotoxic activity against breast cancer cells. Another Portieria hornemannii sample was collected from Noordhoek in the Eastern Cape, it yielded one known compound referred to as 3Z-6-bromo-3-(bromomethylidene)-2,7- dichloro-7-methyloct-1-ene (3.5), as well as one new compound called portieric acid A (3.6) or 5-bromo-2-(bromomethylidene)-6-chloro-6-methylheptanoic acid. Portieric acid A showed slight cytotoxic activity and also represents a new class of compound within the genus Portieria. The isolation of secondary metabolites from the South African red alga, Laurencia glomerata, yielded two known compounds; 7-hydroxylaurene (4.9) and cis-neolaurencenyne (4.12), as well as one chamigrane related compound (4.11). Laurencia flexuosa yielded one known compound called 3Z-bromofucin (4.13). Using 1H NMR, GC and molecular systematics, a novel method for identifying different species of Laurencia was also investigated.
- Full Text:
- Date Issued: 2012
The natural product chemistry of South African Plocamium species
- Authors: Knott, Michael George
- Date: 2003
- Subjects: Marine algae -- South Africa Red algae -- South Africa Green algae -- South Africa Halimeda -- South Africa
- Language: English
- Type: Thesis , Masters , MPharm
- Identifier: vital:3820 , http://hdl.handle.net/10962/d1004920
- Description: The brine shrimp lethality assay was used as a preliminary tool to screen eighteen seaweeds collected from the South African coast. Of the seaweeds tested, the red algae Plocamium corallorhiza and Hypnea rosea, and the green alga Halimeda sp., showed the most potent activity. The chemical investigation of P. corallorhiza resulted in the isolation and structural elucidation of five previously undescribed secondary metabolites, along with three known compounds and four possible artifacts of the extraction process. Standard spectroscopic methods and comparison with known compounds were used to determine the structures of the new metabolites. The new compounds included the linear halogenated monoterpenes 4,8-dibromo-1, 1-dichloro-3,7-dimethyl-2,6-octadiene (99), 4,6-dibromo-l, 1-dichloro-3,7-dimethyl-2,7-octadiene (100), 4,8-dibromo-l, 1,7-trichloro-3,7-dimethyl-2,5-octadiene (101) and 3,4,6,7-tetrachloro-3,7-dimethyl-l-octene (102) and the cyclic monoterpene 5-bromo-5-bromomethyl-I-chlorovinyl-2,4-dichloro-methylcyclohexane (103) while the known compounds were identified as 4-bromo-5-bromomethyl-1chlorovinyl-2,5-dichloro-methylcyclohexane (35), 1,4,8-tribromo-3, 7 -dichloro-3,7-dimethyl-1,5-octadiene (94) and 8-bromo-1,3,4,7-tetrachloro-3,7-dimethyl-1,5-octadiene (96). The four methoxylated compounds (104-107) were presumably formed via a standard substitution reaction between the halogenated monoterpenes 96 and 101 and MeOH, which was used as a component in the extraction solvent. With over 100 000 natural products having been reported, it has become necessary to employ an efficient dereplication strategy to quickly identify known compounds. A simple Gas Chromatography-Mass Spectrometry (GC-MS) method for the efficient physicochemical screening, identification and dereplication of Plocamium metabolites was developed. In this study the crude extracts of P. corallorhiza, P. cornutum and P. maxillosum were screened by GC-MS and the retention times and mass spectral fragmentation patterns of compounds 94, 96, 99 - 107 were used to quickly identify known and new compounds in the crude extracts of P. cornutum and P. maxillosum. This data indicated that compounds 99, 100, 103 were present in both P. corallorhiza and P.cornutum, while compound 102 was found to be present in P. corallorhiza, P. cornutum and P. maxillosum. These studies also indicated that ecotypes and chemotypes are not a significant feature of P. corallorhiza and P. cornutum. Different species of Plocamium (namely: P. corallorhiza, P. cornutum, and P. maxillosum) have very different chemical profiles, and GC may therefore have appreciable taxonomic application in the identification of the different Plocamium spp. which are endemic to South Africa.
- Full Text:
- Date Issued: 2003
- Authors: Knott, Michael George
- Date: 2003
- Subjects: Marine algae -- South Africa Red algae -- South Africa Green algae -- South Africa Halimeda -- South Africa
- Language: English
- Type: Thesis , Masters , MPharm
- Identifier: vital:3820 , http://hdl.handle.net/10962/d1004920
- Description: The brine shrimp lethality assay was used as a preliminary tool to screen eighteen seaweeds collected from the South African coast. Of the seaweeds tested, the red algae Plocamium corallorhiza and Hypnea rosea, and the green alga Halimeda sp., showed the most potent activity. The chemical investigation of P. corallorhiza resulted in the isolation and structural elucidation of five previously undescribed secondary metabolites, along with three known compounds and four possible artifacts of the extraction process. Standard spectroscopic methods and comparison with known compounds were used to determine the structures of the new metabolites. The new compounds included the linear halogenated monoterpenes 4,8-dibromo-1, 1-dichloro-3,7-dimethyl-2,6-octadiene (99), 4,6-dibromo-l, 1-dichloro-3,7-dimethyl-2,7-octadiene (100), 4,8-dibromo-l, 1,7-trichloro-3,7-dimethyl-2,5-octadiene (101) and 3,4,6,7-tetrachloro-3,7-dimethyl-l-octene (102) and the cyclic monoterpene 5-bromo-5-bromomethyl-I-chlorovinyl-2,4-dichloro-methylcyclohexane (103) while the known compounds were identified as 4-bromo-5-bromomethyl-1chlorovinyl-2,5-dichloro-methylcyclohexane (35), 1,4,8-tribromo-3, 7 -dichloro-3,7-dimethyl-1,5-octadiene (94) and 8-bromo-1,3,4,7-tetrachloro-3,7-dimethyl-1,5-octadiene (96). The four methoxylated compounds (104-107) were presumably formed via a standard substitution reaction between the halogenated monoterpenes 96 and 101 and MeOH, which was used as a component in the extraction solvent. With over 100 000 natural products having been reported, it has become necessary to employ an efficient dereplication strategy to quickly identify known compounds. A simple Gas Chromatography-Mass Spectrometry (GC-MS) method for the efficient physicochemical screening, identification and dereplication of Plocamium metabolites was developed. In this study the crude extracts of P. corallorhiza, P. cornutum and P. maxillosum were screened by GC-MS and the retention times and mass spectral fragmentation patterns of compounds 94, 96, 99 - 107 were used to quickly identify known and new compounds in the crude extracts of P. cornutum and P. maxillosum. This data indicated that compounds 99, 100, 103 were present in both P. corallorhiza and P.cornutum, while compound 102 was found to be present in P. corallorhiza, P. cornutum and P. maxillosum. These studies also indicated that ecotypes and chemotypes are not a significant feature of P. corallorhiza and P. cornutum. Different species of Plocamium (namely: P. corallorhiza, P. cornutum, and P. maxillosum) have very different chemical profiles, and GC may therefore have appreciable taxonomic application in the identification of the different Plocamium spp. which are endemic to South Africa.
- Full Text:
- Date Issued: 2003
Gradient high performance liquid chromatographic method for the simultaneous analysis of efavirenz, emtricitabine and tenofovir
- Authors: Koekemoer, Sonya Mariana
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/54679 , vital:26599
- Description: In 2014, approximately 6.8 million people in South Africa were HIV-positive, and the majority of those affected are aged 15 or older. A fixed-dose combination (FDC) antiretroviral (ARV) dosage form containing one non-nucleotide reverse transcriptase inhibitor (efavirenz) and two nucleotide reverse transcriptase inhibitors (emtricitabine and tenofovir) was licensed in South Africa in April 2013. New consolidated guidelines for HIV management and prevention of mother to child transmission (PMTCT) were published by the South African Department of Health in December 2014 and the FDC is now the recommended first-line treatment for HIV-positive patients. According to these guidelines all such people aged 15 and older, and weighing more than 40 kg, with a CD4 count of ≤ 500/ μl will be eligible for antiretroviral therapy (ART) using the FDC. In addition every pregnant and breastfeeding woman is eligible for lifelong ART regardless of CD4 count and EFV can be used as first-line treatment for pregnant women regardless of the length of gestation state of the pregnancy at that time. The use of this simplified regime is likely to promote much needed and improved adherence to therapy. An investigation into the development of a stability-indicating reversed-phase high performance liquid chromatography (RP-HPLC) method for the simultaneous quantitation of EFV, FTC and TNF was undertaken. Isocratic HPLC analysis was found to be unsuitable due to the highly polar FTC molecule eluting in the void. Therefore a gradient HPLC method was developed and validated. The method was validated according to the International Conference on Harmonisation, now known as International Council for Harmonization (ICH). Correlation coefficients > 0.999 were obtained for each assessment of linearity and FTC, TNF and EFV are linear in the range 0.4-40 μg/ml, 0.6-60 μg/ml and 1.2-120 μg/ml. The equation of the best-fit least squares regression lines for FTC, TNF and EFV were y = 0.0191x+0.0007, y = 0.0163x+0.0116 and y = 0.01x+0.016, respectively. The method is accurate as the y-intercept was < 2% of the detector response for all ARV, and the method is precise in terms of intra- and inter-assay precision as all % RSD < 2%. The stability-indicating nature of the method was demonstrated under acidic, alkaline and oxidative stress in addition to UV exposure and elevated temperatures, and the individual chromatograms were overlaid using Empower® 3 Software to establish whether there was interference with the peaks of interest. The forced degradation studies demonstrated the selectivity of the method for the ARV compounds. The method was applied to assay and in vitro dissolution studies of commercially available tablets. The amount of each active ingredient released from Atripla® was determined and compared to the amount of each drug released from Aspen Efavirenz® and Truvada® (a combination of FTC and TNF). The percent FTC released from Atripla® and Truvada® was similar based on the acceptance criteria for immediate-release BCS class 1 compounds. Statistical analysis was undertaken to compare the dissolution profiles of FTC, TNF and EFV. The percent of these compounds released in these studies indicate that bioequivalence testing would be required to declare these products interchangeable. The validated RP-HPLC and in vitro dissolution test method are suitable for routine quality control testing of solid oral dosage forms containing EFV, FTC and TNF, and as the dissolution method can discriminate between different formulations of the same molecule, these tools can also be used for analysis during formulation development studies. The method is not suitable for the analysis of the ARV plasma due to lack of sensitivity and an inability to quantitate the compounds at the required concentration levels. The use of HPLC with mass spectroscopy for quantitation would enhance the sensitivity of the method and may eliminate the quantitation of the molecules in the presence of interference that was observed when using UV detection. Fixed dose combination tablets are convenient for patient therapy and it is likely that in the future more molecules will be formulated into such dosage forms. However formulations such as these can pose significant difficulties when developing and using analytical methods for the quantitation of all compounds in the dosage form at the same time, in particular when the compounds have vastly different physico-chemical properties that impact the quality of a separation and therefore the analysis. Therefore when embarking on the development of FDC product cognisance of the difficulties of developing single methods for the analyses is required and approaches to overcome these difficulties should be considered.
- Full Text:
- Date Issued: 2016
- Authors: Koekemoer, Sonya Mariana
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/54679 , vital:26599
- Description: In 2014, approximately 6.8 million people in South Africa were HIV-positive, and the majority of those affected are aged 15 or older. A fixed-dose combination (FDC) antiretroviral (ARV) dosage form containing one non-nucleotide reverse transcriptase inhibitor (efavirenz) and two nucleotide reverse transcriptase inhibitors (emtricitabine and tenofovir) was licensed in South Africa in April 2013. New consolidated guidelines for HIV management and prevention of mother to child transmission (PMTCT) were published by the South African Department of Health in December 2014 and the FDC is now the recommended first-line treatment for HIV-positive patients. According to these guidelines all such people aged 15 and older, and weighing more than 40 kg, with a CD4 count of ≤ 500/ μl will be eligible for antiretroviral therapy (ART) using the FDC. In addition every pregnant and breastfeeding woman is eligible for lifelong ART regardless of CD4 count and EFV can be used as first-line treatment for pregnant women regardless of the length of gestation state of the pregnancy at that time. The use of this simplified regime is likely to promote much needed and improved adherence to therapy. An investigation into the development of a stability-indicating reversed-phase high performance liquid chromatography (RP-HPLC) method for the simultaneous quantitation of EFV, FTC and TNF was undertaken. Isocratic HPLC analysis was found to be unsuitable due to the highly polar FTC molecule eluting in the void. Therefore a gradient HPLC method was developed and validated. The method was validated according to the International Conference on Harmonisation, now known as International Council for Harmonization (ICH). Correlation coefficients > 0.999 were obtained for each assessment of linearity and FTC, TNF and EFV are linear in the range 0.4-40 μg/ml, 0.6-60 μg/ml and 1.2-120 μg/ml. The equation of the best-fit least squares regression lines for FTC, TNF and EFV were y = 0.0191x+0.0007, y = 0.0163x+0.0116 and y = 0.01x+0.016, respectively. The method is accurate as the y-intercept was < 2% of the detector response for all ARV, and the method is precise in terms of intra- and inter-assay precision as all % RSD < 2%. The stability-indicating nature of the method was demonstrated under acidic, alkaline and oxidative stress in addition to UV exposure and elevated temperatures, and the individual chromatograms were overlaid using Empower® 3 Software to establish whether there was interference with the peaks of interest. The forced degradation studies demonstrated the selectivity of the method for the ARV compounds. The method was applied to assay and in vitro dissolution studies of commercially available tablets. The amount of each active ingredient released from Atripla® was determined and compared to the amount of each drug released from Aspen Efavirenz® and Truvada® (a combination of FTC and TNF). The percent FTC released from Atripla® and Truvada® was similar based on the acceptance criteria for immediate-release BCS class 1 compounds. Statistical analysis was undertaken to compare the dissolution profiles of FTC, TNF and EFV. The percent of these compounds released in these studies indicate that bioequivalence testing would be required to declare these products interchangeable. The validated RP-HPLC and in vitro dissolution test method are suitable for routine quality control testing of solid oral dosage forms containing EFV, FTC and TNF, and as the dissolution method can discriminate between different formulations of the same molecule, these tools can also be used for analysis during formulation development studies. The method is not suitable for the analysis of the ARV plasma due to lack of sensitivity and an inability to quantitate the compounds at the required concentration levels. The use of HPLC with mass spectroscopy for quantitation would enhance the sensitivity of the method and may eliminate the quantitation of the molecules in the presence of interference that was observed when using UV detection. Fixed dose combination tablets are convenient for patient therapy and it is likely that in the future more molecules will be formulated into such dosage forms. However formulations such as these can pose significant difficulties when developing and using analytical methods for the quantitation of all compounds in the dosage form at the same time, in particular when the compounds have vastly different physico-chemical properties that impact the quality of a separation and therefore the analysis. Therefore when embarking on the development of FDC product cognisance of the difficulties of developing single methods for the analyses is required and approaches to overcome these difficulties should be considered.
- Full Text:
- Date Issued: 2016
The study of the potentiation of anticholinergic side effects of tricyclic antidepressives by female sex steroids
- Authors: Kok, Eric Charl
- Date: 1981
- Subjects: Antidepressants , Steroid hormones
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3822 , http://hdl.handle.net/10962/d1005623 , Antidepressants , Steroid hormones
- Description: It has been recorded that women respond to tricyclic antidepressives with a greater incidence of anticholinergic side effects than men do, particularly women taking an exogenous source of oestrogen. The aim of this study was to investigate the influence that ethinyl oestradiol and Premarin© had on the metabolism of a number of tricyclic antidepressives, and also the influence they had on the binding ability of microsomes to imipramine. Rat hepatocyctes and microsomes were used. Detection techniques used were High Pressure Liquid Chromatography and Spectrophotometry respectively. In addition to these studies, a study of the anticholinergic activity of Nomifensine, tricyclic antidepressives and their derivatives was performed on a rat jujenum. Results conclusively showed that ethinyl oestradiol had a marked influence on the metabolism of the tricyclic antidepressives studied. Premarin© had Iittle, if any influence. However, both ethinyl oestradiol and Premarin© affected the binding of microsomes to imipramine, but ethinyl oestradiol had the greater effect. The parent compound in each case exhibited a higher pAZ value. Results indicate that a possible explanation for the increased anticholinergic side effect is due to an inhibition of the metabolism of the tricyclic antidepressives by oestrogen.
- Full Text:
- Date Issued: 1981
- Authors: Kok, Eric Charl
- Date: 1981
- Subjects: Antidepressants , Steroid hormones
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3822 , http://hdl.handle.net/10962/d1005623 , Antidepressants , Steroid hormones
- Description: It has been recorded that women respond to tricyclic antidepressives with a greater incidence of anticholinergic side effects than men do, particularly women taking an exogenous source of oestrogen. The aim of this study was to investigate the influence that ethinyl oestradiol and Premarin© had on the metabolism of a number of tricyclic antidepressives, and also the influence they had on the binding ability of microsomes to imipramine. Rat hepatocyctes and microsomes were used. Detection techniques used were High Pressure Liquid Chromatography and Spectrophotometry respectively. In addition to these studies, a study of the anticholinergic activity of Nomifensine, tricyclic antidepressives and their derivatives was performed on a rat jujenum. Results conclusively showed that ethinyl oestradiol had a marked influence on the metabolism of the tricyclic antidepressives studied. Premarin© had Iittle, if any influence. However, both ethinyl oestradiol and Premarin© affected the binding of microsomes to imipramine, but ethinyl oestradiol had the greater effect. The parent compound in each case exhibited a higher pAZ value. Results indicate that a possible explanation for the increased anticholinergic side effect is due to an inhibition of the metabolism of the tricyclic antidepressives by oestrogen.
- Full Text:
- Date Issued: 1981
Infant health: a community-based assessment and educational intervention in two rural communities in the Eastern Cape.
- Authors: Kuzeeko, Faith
- Date: 2021
- Subjects: Angus Gillis Foundation (South Africa) , Infants -- Mortality -- South Africa , Infants -- Health and hygiene -- South Africa , Breastfeeding -- South Africa -- Eastern Cape , Infants -- Care -- Equipment and supplies , Infants -- Care -- South Africa -- Eastern Cape -- Case studies , Children -- Mortality -- South Africa , High throughput screening (Drug development)
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/170837 , vital:41964
- Description: South Africa is on track to attaining the 2030 Agenda on reducing under-five deaths to 25 deaths per 1000 live births at its current momentum, however an unacceptable amount of infants are still at risk from preventable illnesses. Malnutrition is the major underlined cause of infant mortality rates in South Africa. Exclusive breastfeeding rates are low nationwide due to low exposure to breastfeeding information, some mothers having other commitments and others having breastfeeding difficulties. Implementation and expansion of simple, cost-effective interventions, such as exclusive breastfeeding for six months to reduce and/or prevent infant mortality rates, remains low in South Africa. The aim of the study was to determine one infant health issue of major concern to participants in two rural villages in the Eastern Cape, namely Glenmore and Ndwayana. The identified infant health issue was used to design an educational intervention in the villages. This was the second phase of this study. A community-based participatory approach was utilized in which the Angus Gillis Foundation, a non-profit organization that works in these communities, was one of the stakeholders. Stock status of WHO priority medicines for infants, semi-structured interviews and focus group discussions were carried out during the baseline study. The intervention phase contained pre-2 intervention semi-structured interviews with ten pregnant women followed by an educational intervention with nine out of the ten; and finally, a post-intervention with seven out of the ten women. A questionnaire was completed by members of the Angus Gillis Foundation to provide feedback on the sustainability of the intervention. Semi-structured interviews revealed that medicines stocked at the clinic parallel those indicated in the WHO priority medicines list for infants. The results from the focus group discussions indicated that mothers do not exclusively breastfeed their infants during the first six months. Pre- and post-intervention results on exclusive breastfeeding illustrated a positive change in participants’ knowledge and intent to breastfeed exclusively for six months. They showed a better understanding of the importance of exclusive breastfeeding and indicated a more focussed intention and confidence to carry out optimal breastfeeding practices. In the questionnaire the members of the Angus Gillis Foundation stated that the intervention is sustainable as it was linked with the existing networks. These include educational programs carried out in the villages by the foundation together with positive health champions, community health workers and women self-help groups; which will be able to build on the present knowledge base. Finally, the study also included the design of a booklet on the identified infant health issue. In conclusion, participants highlighted lack of understanding regarding breastfeeding as an issue of concern during the baseline phase of the study. This community-based educational intervention improved the understanding of breastfeeding among the participants, resulting in a positive change in perception with regards to exclusive breastfeeding practices.
- Full Text:
- Date Issued: 2021
- Authors: Kuzeeko, Faith
- Date: 2021
- Subjects: Angus Gillis Foundation (South Africa) , Infants -- Mortality -- South Africa , Infants -- Health and hygiene -- South Africa , Breastfeeding -- South Africa -- Eastern Cape , Infants -- Care -- Equipment and supplies , Infants -- Care -- South Africa -- Eastern Cape -- Case studies , Children -- Mortality -- South Africa , High throughput screening (Drug development)
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/170837 , vital:41964
- Description: South Africa is on track to attaining the 2030 Agenda on reducing under-five deaths to 25 deaths per 1000 live births at its current momentum, however an unacceptable amount of infants are still at risk from preventable illnesses. Malnutrition is the major underlined cause of infant mortality rates in South Africa. Exclusive breastfeeding rates are low nationwide due to low exposure to breastfeeding information, some mothers having other commitments and others having breastfeeding difficulties. Implementation and expansion of simple, cost-effective interventions, such as exclusive breastfeeding for six months to reduce and/or prevent infant mortality rates, remains low in South Africa. The aim of the study was to determine one infant health issue of major concern to participants in two rural villages in the Eastern Cape, namely Glenmore and Ndwayana. The identified infant health issue was used to design an educational intervention in the villages. This was the second phase of this study. A community-based participatory approach was utilized in which the Angus Gillis Foundation, a non-profit organization that works in these communities, was one of the stakeholders. Stock status of WHO priority medicines for infants, semi-structured interviews and focus group discussions were carried out during the baseline study. The intervention phase contained pre-2 intervention semi-structured interviews with ten pregnant women followed by an educational intervention with nine out of the ten; and finally, a post-intervention with seven out of the ten women. A questionnaire was completed by members of the Angus Gillis Foundation to provide feedback on the sustainability of the intervention. Semi-structured interviews revealed that medicines stocked at the clinic parallel those indicated in the WHO priority medicines list for infants. The results from the focus group discussions indicated that mothers do not exclusively breastfeed their infants during the first six months. Pre- and post-intervention results on exclusive breastfeeding illustrated a positive change in participants’ knowledge and intent to breastfeed exclusively for six months. They showed a better understanding of the importance of exclusive breastfeeding and indicated a more focussed intention and confidence to carry out optimal breastfeeding practices. In the questionnaire the members of the Angus Gillis Foundation stated that the intervention is sustainable as it was linked with the existing networks. These include educational programs carried out in the villages by the foundation together with positive health champions, community health workers and women self-help groups; which will be able to build on the present knowledge base. Finally, the study also included the design of a booklet on the identified infant health issue. In conclusion, participants highlighted lack of understanding regarding breastfeeding as an issue of concern during the baseline phase of the study. This community-based educational intervention improved the understanding of breastfeeding among the participants, resulting in a positive change in perception with regards to exclusive breastfeeding practices.
- Full Text:
- Date Issued: 2021
Application of capillary electrophoresis for the assay of erythromycin and its related substance
- Authors: Lalloo, Anita Kantilal
- Date: 1997
- Subjects: Antibiotics -- Analysis , Capillary electrophoresis , Erythromycin -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3765 , http://hdl.handle.net/10962/d1003243 , Antibiotics -- Analysis , Capillary electrophoresis , Erythromycin -- Analysis
- Description: Capillary Electrophoresis (CE) is a high resolution analytical technique that may be employed in the separation and quantification of a wide range of analytes. The enormous efficiency obtained in CE are well suited for complex mixtures in which resolution of a large number of peaks in a short time is desirable. Therefore, CE has a promising future in pharmaC-eutical analysis. The separation mechanism of CE is based on the differential electrophoretic mobility of the solutes inside a buffer filled capillary upon the application of a voltage. Capillary electrophoresis is especially suitable for ionic species. The full potential of this technique can only be realised through the manipulation of numerous experimental parameters. In the present study, a CE method has been developed for the analysis of the macrolide antibiotics: erythromycin, oleandomycin, troleandomycin and josamycin. The selection of initial analysis conditions and optimisation of selectivity are reviewed. A systematic approach to method development was used to maximise analyte differential electrophoretic mobilities, by adjusting the pH. Thereafter, the influences of electrolyte molarity and electrolyte additives were investigated. In addition, some instrumental parameters, such as capillary length emf diameter, applied voltage and injection conditions were varied. The effect of the sample solvent and oncapillary concentration techniques such as FASI, were investigated. Also, the influence of injecting a water plug on the quantity of sample injected was demonstrated. Full resolution was achieved with the addition of methanol to the electrolyte. The applicability of CE for the assay of erythromycin and its related substances was investigated. Two methods were developed and successfully validated using CE: one for the quantitative determination of erythromycin alone and another for erythromycin related substances in the presence of large quantities of erythromycin A. Several related substances and impurities that result from the fermentation process used to produce erythromycin as well as degradation products are known to be present in commercial sa~ples. These impurities include erythromycin B, C, D, E, F, erythromycin enol ether, anhydroerythromycin and N-demethylerythromycin. Currently both the USP and BP official assays for the analysis of erythromycin involve the use of microbiological assays. These methods are limited as they are unable to differentiate between erythromycin and its related substances and degradation products. Furthermore, the microbiological assays are time-consuming and tedious to perform. 11 The CE methods developed for the analysis of erythromycin and for its related substances were fully validated in terms of precision, linearity, accuracy, sensitivity and stability. In addition, erythromycin was subjected to six stress modes and the stressed samples were analysed. An intemal standard was employed to provide acceptable precision for the migration time « 1.80 % RSD) and peak area « 4.44 % RSD). Optimum sensitivity was obtained using low UV wavelengths, with LOO values of less than 10 % for the related substances. The developed method was accurate for erythromycin C, anhydroerythromycin and N-demethylerythromycin, even in the presence of large concentrations of the parent. The method for~ erythromycin related substances was applied to the determination of impurities in three commercial erythromycin bases. The CE methods developed were rapid, precise, specific and stability-indicating and may be used to provide additional information to augment that attained by HPLC for purity assessment and in stability studies of erythromycin. Capillary electrophoresis is a simple, cost-effective technique that is capable of generating high quality data. This technique will become firmly established within pharmaceutical analysis for main peak and related impurity determination assays as familiarity becomes more widespread across the pharmaceutical industry and improvements in instrumentation are performed.
- Full Text:
- Date Issued: 1997
- Authors: Lalloo, Anita Kantilal
- Date: 1997
- Subjects: Antibiotics -- Analysis , Capillary electrophoresis , Erythromycin -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3765 , http://hdl.handle.net/10962/d1003243 , Antibiotics -- Analysis , Capillary electrophoresis , Erythromycin -- Analysis
- Description: Capillary Electrophoresis (CE) is a high resolution analytical technique that may be employed in the separation and quantification of a wide range of analytes. The enormous efficiency obtained in CE are well suited for complex mixtures in which resolution of a large number of peaks in a short time is desirable. Therefore, CE has a promising future in pharmaC-eutical analysis. The separation mechanism of CE is based on the differential electrophoretic mobility of the solutes inside a buffer filled capillary upon the application of a voltage. Capillary electrophoresis is especially suitable for ionic species. The full potential of this technique can only be realised through the manipulation of numerous experimental parameters. In the present study, a CE method has been developed for the analysis of the macrolide antibiotics: erythromycin, oleandomycin, troleandomycin and josamycin. The selection of initial analysis conditions and optimisation of selectivity are reviewed. A systematic approach to method development was used to maximise analyte differential electrophoretic mobilities, by adjusting the pH. Thereafter, the influences of electrolyte molarity and electrolyte additives were investigated. In addition, some instrumental parameters, such as capillary length emf diameter, applied voltage and injection conditions were varied. The effect of the sample solvent and oncapillary concentration techniques such as FASI, were investigated. Also, the influence of injecting a water plug on the quantity of sample injected was demonstrated. Full resolution was achieved with the addition of methanol to the electrolyte. The applicability of CE for the assay of erythromycin and its related substances was investigated. Two methods were developed and successfully validated using CE: one for the quantitative determination of erythromycin alone and another for erythromycin related substances in the presence of large quantities of erythromycin A. Several related substances and impurities that result from the fermentation process used to produce erythromycin as well as degradation products are known to be present in commercial sa~ples. These impurities include erythromycin B, C, D, E, F, erythromycin enol ether, anhydroerythromycin and N-demethylerythromycin. Currently both the USP and BP official assays for the analysis of erythromycin involve the use of microbiological assays. These methods are limited as they are unable to differentiate between erythromycin and its related substances and degradation products. Furthermore, the microbiological assays are time-consuming and tedious to perform. 11 The CE methods developed for the analysis of erythromycin and for its related substances were fully validated in terms of precision, linearity, accuracy, sensitivity and stability. In addition, erythromycin was subjected to six stress modes and the stressed samples were analysed. An intemal standard was employed to provide acceptable precision for the migration time « 1.80 % RSD) and peak area « 4.44 % RSD). Optimum sensitivity was obtained using low UV wavelengths, with LOO values of less than 10 % for the related substances. The developed method was accurate for erythromycin C, anhydroerythromycin and N-demethylerythromycin, even in the presence of large concentrations of the parent. The method for~ erythromycin related substances was applied to the determination of impurities in three commercial erythromycin bases. The CE methods developed were rapid, precise, specific and stability-indicating and may be used to provide additional information to augment that attained by HPLC for purity assessment and in stability studies of erythromycin. Capillary electrophoresis is a simple, cost-effective technique that is capable of generating high quality data. This technique will become firmly established within pharmaceutical analysis for main peak and related impurity determination assays as familiarity becomes more widespread across the pharmaceutical industry and improvements in instrumentation are performed.
- Full Text:
- Date Issued: 1997
Cimetidine as a free radical scavenger
- Authors: Lambat, Zaynab Yusuf
- Date: 2003
- Subjects: Cimetidine , Cimetidine -- Physiological effect , Cimetidine -- Therapeutic use , Alzheimer's disease -- Treatment , Cancer -- Treatment , Free radicals (Chemistry) -- Physiological effect
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3766 , http://hdl.handle.net/10962/d1003244 , Cimetidine , Cimetidine -- Physiological effect , Cimetidine -- Therapeutic use , Alzheimer's disease -- Treatment , Cancer -- Treatment , Free radicals (Chemistry) -- Physiological effect
- Description: The present study was undertaken to determine the effects and possible mechanism of action of cimetidine in cancer and Alzheimer’s disease (AD). Throughout this study emphasis is placed on free radical levels since the magnitude of the relationship between diseases and the levels of free radicals vary from one disease to another. Studies were carried out to examine the effect of cimetidine on free radical levels using superoxide formation and lipid peroxidation as indicators of free radical levels. The experiments revealed that addition of cimetidine, especially in high concentrations (0.5 and 1.0 x10-6 M) significantly inhibited WHCO6 cancer cell growth rather than cancer cell growth, as no normal control was available. Free radical formation as well as hydroxyl radical formation were reduced in the deoxyribose assay. In addition, cimetidine exhibits properties of binding to metals such as copper and iron. To maintain consistency in the experiments, a WHCO6 (Wits Human Carcinoma of the Oesophagus) cell line was used to investigate the effect of cimetidine in cancer. Neurodegeneration was induced in the rat brain using neurotoxins such as cyanide to investigate the relationship between cimetidine in AD. A decrease in cancer cell growth was accompanied by a concomitant decrease in the levels of free radicals and lipid peroxidation, suggesting that the growth-inhibitory effects of cimetidine on WHCO6 cancer cells in vitro may be due to free radical scavenging properties. This proposal was further strengthened by determination of free radical levels in the rat brain. After treatment with neurotoxins to induce neurodegeneration, the levels of free radicals in the rat brain suggest that addition of cimetidine reduces free radical levels in the rat brain in a dosedependent manner. Further experiments were done in an attempt to uncover the underlying mechanism by which cimetidine exhibits free radical scavenging properties. Metal binding studies were done using electrochemical, HPLC and UV/Vis studies. The results show that cimetidine binds iron and copper. These metals have been implicated in free radical production via the Fenton reaction. By binding with cimetidine the metals become unavailable to produce free radicals and hence cimetidine indirectly reduces the formation of free radicals. The final experiment was the determination of cimetidine as a hydroxyl radical scavenger in the deoxyribose assay. Cimetidine was shown to act as a potent hydroxyl radical scavenger, thereby confirming its activity as a free radical scavenger. In addition, cimetidine protects against damage to the deoxyribose sugar, a component of DNA. Whilst there are many theories that explain the therapeutic role of cimetidine in degenerative disease, the actual mechanism of the role of cimetidine is emphasized as a free radical scavenger. Regardless of the mechanism of action, cimetidine does inhibit tumour growth according to this study and also reduce free radical levels in neurodegeneration, which suggests a role for cimetidine as a possible additive in treatment of patients with such disease states. These findings have important clinical implications, and needs to be investigated further.
- Full Text:
- Date Issued: 2003
- Authors: Lambat, Zaynab Yusuf
- Date: 2003
- Subjects: Cimetidine , Cimetidine -- Physiological effect , Cimetidine -- Therapeutic use , Alzheimer's disease -- Treatment , Cancer -- Treatment , Free radicals (Chemistry) -- Physiological effect
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3766 , http://hdl.handle.net/10962/d1003244 , Cimetidine , Cimetidine -- Physiological effect , Cimetidine -- Therapeutic use , Alzheimer's disease -- Treatment , Cancer -- Treatment , Free radicals (Chemistry) -- Physiological effect
- Description: The present study was undertaken to determine the effects and possible mechanism of action of cimetidine in cancer and Alzheimer’s disease (AD). Throughout this study emphasis is placed on free radical levels since the magnitude of the relationship between diseases and the levels of free radicals vary from one disease to another. Studies were carried out to examine the effect of cimetidine on free radical levels using superoxide formation and lipid peroxidation as indicators of free radical levels. The experiments revealed that addition of cimetidine, especially in high concentrations (0.5 and 1.0 x10-6 M) significantly inhibited WHCO6 cancer cell growth rather than cancer cell growth, as no normal control was available. Free radical formation as well as hydroxyl radical formation were reduced in the deoxyribose assay. In addition, cimetidine exhibits properties of binding to metals such as copper and iron. To maintain consistency in the experiments, a WHCO6 (Wits Human Carcinoma of the Oesophagus) cell line was used to investigate the effect of cimetidine in cancer. Neurodegeneration was induced in the rat brain using neurotoxins such as cyanide to investigate the relationship between cimetidine in AD. A decrease in cancer cell growth was accompanied by a concomitant decrease in the levels of free radicals and lipid peroxidation, suggesting that the growth-inhibitory effects of cimetidine on WHCO6 cancer cells in vitro may be due to free radical scavenging properties. This proposal was further strengthened by determination of free radical levels in the rat brain. After treatment with neurotoxins to induce neurodegeneration, the levels of free radicals in the rat brain suggest that addition of cimetidine reduces free radical levels in the rat brain in a dosedependent manner. Further experiments were done in an attempt to uncover the underlying mechanism by which cimetidine exhibits free radical scavenging properties. Metal binding studies were done using electrochemical, HPLC and UV/Vis studies. The results show that cimetidine binds iron and copper. These metals have been implicated in free radical production via the Fenton reaction. By binding with cimetidine the metals become unavailable to produce free radicals and hence cimetidine indirectly reduces the formation of free radicals. The final experiment was the determination of cimetidine as a hydroxyl radical scavenger in the deoxyribose assay. Cimetidine was shown to act as a potent hydroxyl radical scavenger, thereby confirming its activity as a free radical scavenger. In addition, cimetidine protects against damage to the deoxyribose sugar, a component of DNA. Whilst there are many theories that explain the therapeutic role of cimetidine in degenerative disease, the actual mechanism of the role of cimetidine is emphasized as a free radical scavenger. Regardless of the mechanism of action, cimetidine does inhibit tumour growth according to this study and also reduce free radical levels in neurodegeneration, which suggests a role for cimetidine as a possible additive in treatment of patients with such disease states. These findings have important clinical implications, and needs to be investigated further.
- Full Text:
- Date Issued: 2003
The influence of a methylated-β-Cyclodextrin on the solubility and photostability of midazolam in aqueous solution
- Authors: Lebete, Mosimotsana Leah
- Date: 2001 , 2013-04-26
- Subjects: Midazolam -- Solubility
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3823 , http://hdl.handle.net/10962/d1005923 , Midazolam -- Solubility
- Description: Midazolam, used clinically as an anticonvulsant, anxiolytic, muscle relaxant and sedative is a photolabile imidazo-benzodiazepine which is marketed under the trade names Dormicum® and Hypnovel® as tablets and injectables. Because of an aqueous solubility of < 0.1 mg/ml above pH 4, the preparation of aqueous dosage formulations near physiological pH, requires a solubilizer. The aim of this study was thus to prepare a 10 mg/ml midazolam aqueous solution for topical application using randomly-methylated-pcyclodextrin (RAMEB), considered to be a suitable candidate as a solubilizer because of its absorption enhancing properties, and to investigate its effect on both the solubility and the photostability of midazolam. Solubility studies of midazolam (excess of 15 mg/ml) in the presence of 0, 5,10,20, 30% m/v of RAMEB at pH 5.0 and pH 5.8 (phosphate buffer) were undertaken and the results analyzed using a UV method validated for linearity, accuracy, precision and specificity. A stability-indicating HPLC method was developed and validated (precision and accuracy, linearity, range, limit of quantitation, specificity, robustness and ruggedness) for application to kinetic photostability studies and the identification of photodegradants by LC-MS. Forced degradation studies were carried out at concentrations of 0.5 mg/ml of midazolam instead of the target concentration of 10 mg/ml because of the acceleratory effect of the decreased concentration on the rate of photodegradation. The solutions of midazolam with and without RAMEB were irradiated at 550 W/m² for 12 hrs in order to degrade the drug to ± 10% of the original concentration. The UV method proved to be valid in terms of linearity with a correlation coefficient of 0.9998, precise and accurate, and specific for the determination of midazolam in the presence of RAMEB. The results of the phase solubility studies indicated that desired solubility of 10 mg/ml was achieved with 30% m/v RAMEB at pH 5.0. RAMEB slightly decreased the photostability of midazolam, the rate constants being 0.137 and 0.154 hr⁻¹ in the absence and presence of RAMEB, respectively. LC-MS analysis revealed that one of the major photoproducts in the presence and absence of RAMEB was N-desalkylflurazepam, a starting material in the synthesis of midazolam. RAMEB inhibited formation of some photoproducts and introduced two new photoproducts, a dimer and an addition product. The difference in the nature of these photoproducts formed may be attributed to the ability of RAMEB to provide conformational control and to stabilize free radicals. Although RAMEB improved the solubility of midazolam to the target concentration, photostability is decreased with the presence of different photoproducts. These studies have however provided information on the overall photostability of midazolam, the identity of its photodegradants and the photodegradation pathway in the presence and absence of RAMEB, and may be used for further method development and validation for the analysis of aqueous dosage forms containing RAMEB as a solubilizer. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 2001
- Authors: Lebete, Mosimotsana Leah
- Date: 2001 , 2013-04-26
- Subjects: Midazolam -- Solubility
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3823 , http://hdl.handle.net/10962/d1005923 , Midazolam -- Solubility
- Description: Midazolam, used clinically as an anticonvulsant, anxiolytic, muscle relaxant and sedative is a photolabile imidazo-benzodiazepine which is marketed under the trade names Dormicum® and Hypnovel® as tablets and injectables. Because of an aqueous solubility of < 0.1 mg/ml above pH 4, the preparation of aqueous dosage formulations near physiological pH, requires a solubilizer. The aim of this study was thus to prepare a 10 mg/ml midazolam aqueous solution for topical application using randomly-methylated-pcyclodextrin (RAMEB), considered to be a suitable candidate as a solubilizer because of its absorption enhancing properties, and to investigate its effect on both the solubility and the photostability of midazolam. Solubility studies of midazolam (excess of 15 mg/ml) in the presence of 0, 5,10,20, 30% m/v of RAMEB at pH 5.0 and pH 5.8 (phosphate buffer) were undertaken and the results analyzed using a UV method validated for linearity, accuracy, precision and specificity. A stability-indicating HPLC method was developed and validated (precision and accuracy, linearity, range, limit of quantitation, specificity, robustness and ruggedness) for application to kinetic photostability studies and the identification of photodegradants by LC-MS. Forced degradation studies were carried out at concentrations of 0.5 mg/ml of midazolam instead of the target concentration of 10 mg/ml because of the acceleratory effect of the decreased concentration on the rate of photodegradation. The solutions of midazolam with and without RAMEB were irradiated at 550 W/m² for 12 hrs in order to degrade the drug to ± 10% of the original concentration. The UV method proved to be valid in terms of linearity with a correlation coefficient of 0.9998, precise and accurate, and specific for the determination of midazolam in the presence of RAMEB. The results of the phase solubility studies indicated that desired solubility of 10 mg/ml was achieved with 30% m/v RAMEB at pH 5.0. RAMEB slightly decreased the photostability of midazolam, the rate constants being 0.137 and 0.154 hr⁻¹ in the absence and presence of RAMEB, respectively. LC-MS analysis revealed that one of the major photoproducts in the presence and absence of RAMEB was N-desalkylflurazepam, a starting material in the synthesis of midazolam. RAMEB inhibited formation of some photoproducts and introduced two new photoproducts, a dimer and an addition product. The difference in the nature of these photoproducts formed may be attributed to the ability of RAMEB to provide conformational control and to stabilize free radicals. Although RAMEB improved the solubility of midazolam to the target concentration, photostability is decreased with the presence of different photoproducts. These studies have however provided information on the overall photostability of midazolam, the identity of its photodegradants and the photodegradation pathway in the presence and absence of RAMEB, and may be used for further method development and validation for the analysis of aqueous dosage forms containing RAMEB as a solubilizer. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 2001
Applicability of a health literacy test from the U.S. in a South African population
- Lecoko, Motlalepule Lebogang Elizabeth
- Authors: Lecoko, Motlalepule Lebogang Elizabeth
- Date: 2001 , 2013-04-29
- Subjects: Literacy , Literacy -- South Africa , Literacy -- Ability testing , Reading -- Ability testing
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3824 , http://hdl.handle.net/10962/d1005931 , Literacy , Literacy -- South Africa , Literacy -- Ability testing , Reading -- Ability testing
- Description: This thesis investigates the suitability and applicability of a health literacy test from the U.S. in a black, Xhosa-speaking, South African population. The concept of literacy is a controversial one which has been much debated, as it is not easy to classifY people as simply either literate or illiterate. As a result there are a number of definitions of literacy that vary with purpose and culture, but the most common one is that a person is literate if he/she can read and write. Estimating literacy from years of schooling is an inexpensive method but is also unreliable, since people generally read 3 to 5 grades below their stated educational level. This method affords little insight into the ability of patients to adequately function in a health care enviromnent, an ability which is referred to as functional health literacy. A number of health literacy tests such as the REALM (Rapid Estimate of Adult Literacy in Medicine) test have been developed to assess this skill. The REALM test is a word recognition test which places people into a relevant grade range estimate according to the number of words pronounced correctly. It appears to assume understanding of the word if the person is able to read that word correctly. In this project 125 black Xhosa-speaking respondents of varying educational levels who were literate in English were interviewed with the aid of an interpreter. Comprehensive demographic data were collected. Respondents were first asked to read all 66 words aloud during which time pronunciation was checked, and thereafter they were asked to explain each word. It was found that the ability to automatically decode and read the words did not necessarily guarantee comprehension of these words. Many of the words proved to be unfamiliar to the majority of the Xhosa respondents who were able to pronounce them correctly, but could not explain them. These tended to be phonetically transparent words which were therefore more accessible to the unfamiliar reader. This research has proven to be of great value in helping identify such words which should be substituted with simpler words for use in health information materials. A number of words could neither be pronounced nor understood by the population majority and, interestingly, a small group of words could not be pronounced but were satisfactorily explained by some respondents. The results showed an extremely poor correlation between the stated educational level and the REALM grade range estimate. This emphasizes the inappropriateness of years of formal schooling as an indicator of functional health literacy. The criteria were established for deciding cases in which the REALM test could be applied (or succeeds) and when it is inapplicable (or fails). It was found to be inapplicable in 41% of cases which clearly indicates that, in its current form, it is not a valid, reliable test to use in determining health literacy in this English second language population. It can, however, be used as a basis fur the development of a more appropriate test. Recommendations for future research direction are presented and an alternative structure for a health literacy test is suggested. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 2001
- Authors: Lecoko, Motlalepule Lebogang Elizabeth
- Date: 2001 , 2013-04-29
- Subjects: Literacy , Literacy -- South Africa , Literacy -- Ability testing , Reading -- Ability testing
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3824 , http://hdl.handle.net/10962/d1005931 , Literacy , Literacy -- South Africa , Literacy -- Ability testing , Reading -- Ability testing
- Description: This thesis investigates the suitability and applicability of a health literacy test from the U.S. in a black, Xhosa-speaking, South African population. The concept of literacy is a controversial one which has been much debated, as it is not easy to classifY people as simply either literate or illiterate. As a result there are a number of definitions of literacy that vary with purpose and culture, but the most common one is that a person is literate if he/she can read and write. Estimating literacy from years of schooling is an inexpensive method but is also unreliable, since people generally read 3 to 5 grades below their stated educational level. This method affords little insight into the ability of patients to adequately function in a health care enviromnent, an ability which is referred to as functional health literacy. A number of health literacy tests such as the REALM (Rapid Estimate of Adult Literacy in Medicine) test have been developed to assess this skill. The REALM test is a word recognition test which places people into a relevant grade range estimate according to the number of words pronounced correctly. It appears to assume understanding of the word if the person is able to read that word correctly. In this project 125 black Xhosa-speaking respondents of varying educational levels who were literate in English were interviewed with the aid of an interpreter. Comprehensive demographic data were collected. Respondents were first asked to read all 66 words aloud during which time pronunciation was checked, and thereafter they were asked to explain each word. It was found that the ability to automatically decode and read the words did not necessarily guarantee comprehension of these words. Many of the words proved to be unfamiliar to the majority of the Xhosa respondents who were able to pronounce them correctly, but could not explain them. These tended to be phonetically transparent words which were therefore more accessible to the unfamiliar reader. This research has proven to be of great value in helping identify such words which should be substituted with simpler words for use in health information materials. A number of words could neither be pronounced nor understood by the population majority and, interestingly, a small group of words could not be pronounced but were satisfactorily explained by some respondents. The results showed an extremely poor correlation between the stated educational level and the REALM grade range estimate. This emphasizes the inappropriateness of years of formal schooling as an indicator of functional health literacy. The criteria were established for deciding cases in which the REALM test could be applied (or succeeds) and when it is inapplicable (or fails). It was found to be inapplicable in 41% of cases which clearly indicates that, in its current form, it is not a valid, reliable test to use in determining health literacy in this English second language population. It can, however, be used as a basis fur the development of a more appropriate test. Recommendations for future research direction are presented and an alternative structure for a health literacy test is suggested. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
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- Date Issued: 2001