Phenylpropanolamine (PPA), a synthetic sympathomimetic amine structurally related to ephedrine has been widely used over t he past 40 years as a nasal decongestant and appetite suppressant. It has been the focus of much controversy concerning the efficacy of the drug in its use as an anorectic agent, and due to the side effects caused by the higher doses of PPA required for appetite suppression. Although extensively used, there is little information concerning the determination of PPA in biological fluids and on the pharmacokinetics of this drug. An adaptation of a published high-performance liquid chromatographic (HPLC) assay for PPA in serum and urine using U.V. detection at 210 nm is presented. PPA was separated in the reversed phase mode. The method has a limit of sensitivity of 5.0 ng/mL and 10.0 ng/mL in serum and urine respectively. Serum concentration data following a single 25 mg dose of phenylpropanolamine in human volunteers demonstrate the application of the analytical method for bioavailability and pharmacokinetic studies. After the administration of 25 mg, 50 mg or 100 mg PPA.HCl solutions to 5 human volunteers, a dose proportionality study demonstrated that PPA appears to exhibit linear kinetics. Linear one body compartment kinetics were assumed and the wagner-Nelson method used to transform in vivo serum data to absorption plots. The serum data were fitted to a model using nonlinear regression techniques to characterize the pharmacokinetic processes of PPA. The absorption of phenylpropanolamine appears to be discontinuous and the drug seems to favour a two body compartment model. The pharmacokinetic parameters obtained from a steady state study using multiple dosing of PPA.HCl solutions compared with those found from previous studies after the administration of sustained-release formulations. A plasma protein binding study using equilibrium dialysis demonstrated that PPA is not highly protein bound in the blood.

Protein A (ProA) chromatography is a bioseparations technique employed throughout the biopharmaceutical industry for the selective capture and purification of IgG-class monoclonal antibodies (mAbs) and Fc-fusion proteins. The rapid growth of mAbs as commercial therapeutics has motivated the need for improved, efficient, and high-throughput purification processes during manufacturing. In direct response, the work presented in thesis aims to 1) increase the scientific community’s understanding of IgG adsorption behavior on ProA chromatography resins and 2) improve the performance of ProA chromatography with ligands that are chemically modified using polyethylene glycol (PEGylated). The results of this thesis suggest that IgG molecules of varying binding strength, or varying elution pH, are capable of competing for binding sites on ProA chromatography resins in simultaneous or sequential adsorption. The competitive phenomenon derives from variance in IgG binding strength, or IgG elution pH, due to differences in sub-class behavior as well as secondary IgG binding interactions with the ProA ligand. Competition is readily apparent in the adsorption of human polyclonal IgG, which has a wide variety of IgG sub-classes and binding epitopes. Additionally, the results presented in this thesis suggest that ProA chromatography resins with PEGylated ligands are a viable path to increase resin robustness and real-world chromatographic selectivity. It is demonstrated that ligand PEGylation can increase resistance to proteolytic digestion, mitigate impurity interactions with mAbs that are bound to ProA, and increase process selectivity against Chinese Hamster Ovary host cell proteins by up to 37%. However, resins with large volumes of conjugated PEG significantly decrease IgG static binding capacity and decrease the available pore space for diffusion, resulting in losses in dynamic binding capacity and productivity. Lighter modifications appear to avoid losses in dynamic binding capacity, however, they do not appear to be effective at mitigating impurity interactions with mAbs that are bound to ProA, which is key to increasing process selectivity. PEGylation of ProA also universally increases the elution pH of IgG molecules by weakening the binding interaction. This last result opens another path of viability for PEGylated ProA ligands for purification of mAbs of Fc-fusion proteins that are sensitive to low pH environments.

M.Sc. / Aerosol samples were collected at stations using simple filter unit, loaded with a 4 7 mm diameter pore size Nuclepore membrane filter. The sampling duration was 24 hours for each sample, with the total of 11 samples: seven for Thohoyandou and four for Bethlehem. The samples were analyzed with energy dispersive X-ray fluorescence spectrometry for up to 20 elements. Comparison for elemental concentrations of the data showed that the samples fall into three clusters on average (major: Ca, Cl, Si. S, Fe and AI; intermediate: K, Ti, Pb, Brand Zn; minor: Mn, Cr, As, Sr, Ni and Cu), with the samples collected from Bethlehem having high concentrations in all clusters. High concentrations in the samples collected on Bethlehem were attributed to entrainement of dust particles during the dry winter period. The results also show that the elements are associated with anthropogenic emissions. Therefore, the sampling station at Thohoyandou is considered as a background station due to the low aerosol concentration. The aerosol concentration levels at Thohoyandou can be attributed to biogenic sources because of the large presence of large forest areas in the region and in the vicinity of the sampling location. The samples collected in both regions reveal highly enriched sulphur, chlorine, zinc and lead. It is clearly proved that these elements come from local soil. Chromium and copper were both slightly enriched in Thohoyandou, but the concentration data for these two elements are not available. However it is supposed that these elements come from local soil as well. In another aerosol analysis, passive (diffusive) samplers were used to measure concentration levels for sulphur dioxide and nitrogen dioxide. The measurement campaign was carried out from Soweto in Johannesburg and Bohlokong in Bethlehem with the campaign lasting for two months during the w~nter season. A very clear result derived for sulphur dioxide was the dominating source contribution from use of coal for heating and cooking in both areas. For nitrogen dioxide, it was found that contribution from traffic in highly populated areas and from industrial activities in the neighbourhood of the two areas was the source. The role played by wood burning, is also another additional source used for domestic heating and even cooking.

No description available.

Tuberculosis (TB) is a highly contagious disease caused by the bacterium known as Mycobacterium tuberculosis which is widely spread in South Africa, especially in the rural areas of the Western Province. Rifampicin, isoniazid and pyrazinamide are the three most effective drugs against this organism. However, most of the current commercial anti-TB formulations are inconvenient to administrate. This results in patient non-compliance which has increased with incomplete tuberculosis treatment and further has intensified the mortality rate. The matter is especially severe amongst the paediatric and geriatric patients. Therefore, creating a "user-friendly" but non-alcoholic liquid formulation should improve the whole situation. The key to a successful formulation relies on sufficient concentrations of the drugs within the formulation together with acceptable stability of these drugs. Therefore, during the pre-formulation stage, the solubility and stability studies of rifampicin, isoniazid and pyrazinamide are to be conducted. Rifampicin, isoniazid and pyrazinamide were fully characterized and identified by means of spectroscopic and thermal techniques. A HPLC method for simultaneous analysis of the three drugs was developed and validated. This HPLC method was employed for all the solubility and stability assessments. Unbuffered HPLC water of pH value 7.01 was chosen as the aqueous solvent. This was decided after the stability of rifampicin, isoniazid and pyrazinamide was studied at a pH range of 2 to 10. The solubility and the stability studies of rifampicin, isoniazid, pyrazinamide, rifampicin with isoniazid, rifampicin with pyrazinamide, isoniazid with pyrazinamide and rifampicin combined with both isoniazid and pyrazinamide were performed in the presence of various agents. These agents can be categorized into three groups: the surfactants (poloxamer 188, poloxamer 407 and sorbitol) which could increase the intrinsic solubility or the drugs by altering the surface tensions of the aqueous solution medium, the suspending agents (carbopol 934 and carbopol 974P) which could enable the amount of dosage required to be homogeneously suspended in the formulation without considering the low intrinsic solubility factor of the drugs, and the complexing agents (ß-cyclodextrin, hydroxypropyl-ß-cyclodextrin and -cyclodextrin) which could initiated the inclusion complex between the host cyclodextrin and the drugs, thus further enhance the solubility of the drugs . The stability assessments were performed after 7-days stability trail with the HPLC method developed. Each drug/combination of drugs were stored in closed ampoules and subjected to 25, 40 and 60° C with or without nitrogen flushing while in the presence of the above mentioned agents. While assessing the solubility/stability of the drugs in the presence of the above mentioned surfactants, the phase-solubility curves indicate that both rifampicin and pyrazinamide fail to achieve the desired concentration. Moreover, the stability-time plots clearly indicate that these surfactants fail to enhance the general stabilities of the drugs. When the stabilizing effects of the above mentioned suspending agents were investigated, it was found that although the desired concentration could be easily accomplished by suspending the drug in the aqueous carbopol solutions, the stabilities of the different drug combinations were still below the regulatory level. Cyclodextrins are well known to form inclusion complexes with less polar drug molecules. The inclusion complexation could enhance both the solubility and the stability of the included drug molecules. The computer force field generated models of the cyclodextrin-drug were used to predict the complexation sites. The results indicated the all the inclusion complexation between the drugs and the cyclodextrins were favourable, but do not necessary protect the potential degradation sites of the drugs. The stability results confirmed the above findings as the cyclodextrins did not enhance the stability of the drugs. Various drug-drug interaction pathways were also predicted from the experimental observations which clearly indicated the stability reductions of these drugs in combination. This leads to the conclusion that a liquid formulation combining rifampicin, isoniazid and pyrazinamide should not initiate the use of aqueous solutions as the protic ions of the solution are able to initiate the degradation of these drugs.

From Introduction: Buchu leaf is a very widely used household medicine in South Africa, and is usually administered in the form of a brandy tincture or a vinegar, known as "buchu brandy" and "buchu vinegar" respectively. These preparations have a great reputation in curing diseases of the kidney and urinary tract, and in addition are employed as local applications to bruises, and for the relief of rheumatic pains. We owe its introduction into medicine to the Hottentot, who gave the name "buchu" or "bookoo" to any aromatic herb or shrub which they found suitable for use as a dusting powder.

Please read the abstract on page 4 of this document / Dissertation (MSc (Chemistry))--University of Pretoria, 2007. / Chemistry / MSc / unrestricted

The metabolism of absorbed xenobiotic compounds in humans results in a mixture of target compounds applicable for analysis, trapped in complex biological matrices. Gas chromatography-mass spectrometry (GC-MS) is a powerful analytical technique that has been successfully applied in the analysis of volatile and semi-volatile compounds from complex biological samples. This is due to the ability of GC-MS to separate different sample constituents at trace levels while providing accurate molecular structural information for the resolved compounds. The complexity of biological specimens and their largely aqueous nature, combined with the physicochemical properties of target analytes resulting from metabolism, greatly precludes direct analysis of biosamples by GC-MS. Traditionally, highly laborious and time consuming sample preparation procedures are performed to isolate and chemically alter target analytes to attain suitable amenity for the detection system. Furthermore, routine analytical procedures in clinical toxicology laboratories are signified by short specimen turn-around times. The commonplace use of GC-MS in modern-day laboratories still suffer from prolonged turn-around times that result from both sample preparation steps and lengthy instrumental analysis. Simplified and cost-effective analytical procedures capable of extracting multiple analytes, with divergent functional groups, from biological matrices in a timely manner are therefore required. To address this issue, this work describes the development of validated extractive-derivatization methods combined with fast GC-MS analysis for expedient and accurate quantitation of different analytes in occupational monitoring and workplace drug testing. Extractive alkylation of acidic analytes phenol, o-cresol, mandelic acid, hippuric acid, and (o-, m-, p-) methylhippuric acid for simultaneous urinary bio-monitoring of occupational exposure to benzene, toluene, ethylbenzene, and xylene, respectively, is performed. Extractive acylation for simultaneous urinary confirmation of basic analytes amphetamine, methamphetamine, norephedrine, methcathinone, ephedrine, methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA), methylenedioxyethylamphetamine (MDEA) and N-methyl-1-(3,4 methylenedioxyphenyl)-2-butanamine (MBDB) in workplace drug testing is performed. The successful combination of abovementioned techniques alongside fast GC-MS allows increased sample throughput and decreased turn-around time for routine analysis while maintaining bioanalytical quantitative criteria, as required in a clinical toxicology laboratory setting. / Dissertation (MSc)--University of Pretoria, 2009. / Chemical Pathology / unrestricted

Orientador: Helena Teixeira Godoy / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-04T01:44:03Z (GMT). No. of bitstreams: 1 Pallone_JulianaAzevedoLima_D.pdf: 1435689 bytes, checksum: 62750d8ae611c0a4dffee7b6395ff256 (MD5) Previous issue date: 2005 / Resumo: Apesar das recentes descobertas a respeito das possíveis ações benéficas dos folatos na saúde humana, ainda são poucos os dados a respeito dos reais teores dessa vitamina nos alimentos brasileiros, principalmente para os vegetais, considerados as principais fontes desse nutriente. Mais escassos ainda são os estudos sobre a estabilidade dos folatos após o cozimento doméstico e o processamento industrial de alimentos. Considerando-se que a concentração de folatos em alimentos é bastante dependente das condições edafoclimáticas, torna-se relevante avaliar essa vitamina em vegetais brasileiros, a fim de se assegurar a ingestão dos folatos pela população. Assim sendo, este trabalho teve como objetivo principal contribuir para o estudo de folatos em vegetais considerados ricos nessa vitamina. Para a determinação dos folatos, utilizou-se a técnica de cromatografia líquida de alta. O método foi validado e aplicado para amostras de espinafre, brócolis, tomate, catchup, molho e suco de tomate, com pequenas adaptações na metodologia referentes à etapa de extração das amostras e no sistema de separação cromatográfica. De forma geral, os parâmetros de validação estabelecidos, para os produtos analisados, foram de 94% para recuperação, coeficiente de variação inferior a 1,4% nos ensaios de repetibilidade e 5, 30, 30, 7pg/mL e 5ng/mL foram os limites de detecção estabelecidos para 5-metiltetraidrofolato (5-metilTHF), 5-formiltetraidrofolato (5-formilTHF), 10-formiltetraidrofolato (10-formilTHF), tetraidrofolato (THF) e 10-metiltetraidrofolato (10-metilTHF), respectivamente. Em tomate e produtos derivados o único folato encontrado foi o 5-metilTHF. Não se observou diferença significativa, entre os teores dessa forma da vitamina, encontrados em tomates cultivados de forma tradicional (entre 176,0 a 326,9mg/100) e orgânica (208,7 a 345,1mg/100g). Nos produtos de tomate verificou-se que a concentração de 5-metilTHF variou significativamente, tanto entre marcas, quanto entre lotes analisados para cada produto. Para catchup, molho e suco de tomate os valores obtidos variaram entre 58,8 e 265,9mg/100g, 103,9 e 325,0mg/100g e 138,9 e 330,8mg/100g, respectivamente. Assim, tanto o tomate como os seus produtos derivados podem ser considerados fontes de folatos. No brócolis além do 5-metilTHF, o 5-formilTHF também foi detectado. Nesse caso, observou-se diferença significativa entre os teores de 5-metilTHF obtidos para o cultivo orgânico (954,0 a 1740,0mg/100g para 5-metilTHF e 7,9 a 13,6mg/100g para 5-formilTHF) e tradicional (414,0 a 742,0mg/100g para 5-metilTHF e 4,8 a 12,8mg/100g para 5-formilTHF), tendo a forma orgânica apresentado as maiores concentrações. No caso do 5-formilTHF não se observou diferença significativa devida ao cultivo. Verificou-se, também, que o brócolis cozido em água, apresentou perdas de aproximadamente 68%, sendo que a maior parte da vitamina (53%) ficou na água de cozimento, indicando ser o processo de lixiviação o maior responsável pela perda dos folatos. Para o espinafre as mesmas formas de folatos encontradas em brócolis (5-metilTHF e 5-formilTHF) foram detectadas. Tanto para brócolis, quanto para espinafre, o 5-metilTHF representou aproximadamente 99% do teor total de folatos. Verificou-se, entretanto, em espinafre, não haver diferença significativa entre os teores de folatos obtidos por cultivo tradicional (386,0 a 550,4mg/100g para 5-metilTHF e 4,0 a 6,3mg/100g para o 5-formilTHF) e orgânico (295,3 a 599,9mg/100g para 5-metilTHF e 5,0 a 7,7mg/100g para o 5-formilTHF). As concentrações das diferentes formas de folatos, em espinafres colhidos em diferentes épocas do ano (setembro a novembro/2002 e maio e julho de 2003) também foram avaliados e revelaram não haver diferença significativa para os valores obtidos ao longo do período estudado. O cozimento em água resultou em perdas de aproximadamente 79% para o 5-metilTHF e 66% para o 5-formilTHF. Pela determinação dos teores de folatos na água de cozimento, concluiu-se que, também para esse caso, as perdas se deram, principalmente, devido a lixiviação. Os resultados obtidos permitiram, potanto, concluir-se que todos os vegetais avaliados no trabalho podem ser considerados importantes fontes de folatos, entretanto, o cozimento em água ocasionou enormes perdas da vitamina. Dessa forma, recomenda-se o aproveitamento da água de cozimento como forma de garantir a ingestão desse importante nutriente / Abstract: Despite the recent discoveries about the possible beneficial actions of folates to human health, data on the real contents of this vitamin in Brazilian foods, are little, ever for vegetables, considered the main source of this nutrient. Scarcer still are studies on the stability of folates after domestic cooking and industrial food processing. Considering that the concentration of folates in foods is somewhat dependent on the soil/climate conditions, it becomes important to evaluate this vitamin in Brazilian vegetables, in order to guarantee the ingestion of folates by the population. Thus the main of this study was to contribute to the knowledge of folates in foods. Folates were determined by high performance liquid chromatography technique. The method was validated and applied to samples of spinach, broccoli, tomatoes, ketchup, sauce and tomato juice. Small adaptations in the methodology were made for the extraction step of the samples and others in the chromatographic separation system. In general, the validation parameters established for the products analyzed were of 94% for recovery, variation coefficient below 1.4% in the repeatability assays and detection limits of 5, 30, 30, 7pg/mL and 5ng/mL 5-methyl-tetrahydrofolate (5-methylTHF), 5-formyl-tetrahydrofolate (5-formylTHF), 10-formyl- tetrahydrofolate (10-formylTHF), tetrahydrofolate (THF) and 10-methyl- tetrahydrofolate (10-methylTHF), respectively. For tomatoe and tomato products the only folate form was 5-methylTHF. There was no significant difference between the contents of this form of the vitamin found in tomatoes cultivated by the traditional (176,0 to 326,9µg/100g) and the organic method (208.7 to 345.1µg/100g) form. For tomatoe products, both, brands and batches showed significant differences in the concentration of 5-methylTHF. For ketchup, sauce and tomato juice the values were from 58.8 to 265.9µg/100g, 103.9 to 325.0µg/100g and 138.9 to 330.8µg/100g, respectively. Thus, tomatoes and tomato products can be considered as folates sources. In broccolis, in addition to 5-methylTHF, 5-formylTHF was also detected. In this case, these was a significant difference in the contents of 5-methylTHF for the organic (954.0 to 1740.0µg/100g for 5-methylTHF and 7.9 to 13.6µg/100g for 5-formylTHF) and traditional (414.0 to 742.0µg/100g for 5-methylTHF and 4.8 to 12.8µg/100g for 5-formylTHF) cultivation, the organic form presenting higher concentrations. For 5-formylTHF there was not significant difference. It was also noted, that when the broccoli was cooked in water, the losses were of approximately 85%, that most of the vitamin (70%) being founder the cooking water, indicating that the leaching process is responsible for the greater losses of folates. For the spinach the same folates forms were found as in broccolis (5-methylTHF and 5-formylTHF). For both, broccoli and spinach, 5-methyllTHF corresponds to approximately 99% of the folates contents. For spinach there was no significant difference in the folates contents for traditional (386.0 to 550.4µg/100g for 5-methylTHF and 4.0 to 6.3µg/100g for 5-formylTHF) and organic (295.3 to 599.9µg/100g for 5-methylTHF and 5.0 to 7.7µg/100g for 5-formylTHF) cultivation. The contents of the different folate forms, in spinach were also evaluated at different periods of the year (September to November/2002 and May to July/2003), and it was concluded there was no significant difference between the values obtained in the periods studied. Cooking in water resulted in losses of 66% for 5-methylTHF and 47% for 5-formylTHF. The determination of folates in the cooking water, confirmed that, in this case, the losses were mostly due to leaching, most of the vitamin, being found in the water. It was concluded that all the vegetables evaluated in this study could be considered important folates sources, however, the cooking in water caused enormous losses of the vitamin. It is therefore of great importance to use the cooking water so that adequate amounts of this important nutrient are ingested. / Doutorado / Ciência de Alimentos / Doutor em Ciência de Alimentos

A novel chromatographic method was devised that makes use of the superb group separation power of normal phase supercritical fluid chromatography (SFC) combined with a fast second separation by a resistively heated gas chromatograph (GC). The SFC was operated isothermally with stopped flow to provide the time required for the GC analysis. The GC analysis had a typical cycle time of 1 minute. During this time the GC column was independently heated at a rate of 450°C/min to 250°C and actively cooled down again to -50°C before the next GC injection takes place. This was achieved with an in-house designed, resistively heated, temperature programmable gas chromatograph. Various temperature measurement circuits were also evaluated. An interface was developed that allows transfer between the SFC and the GC in such a way that the entire eluent from the first separation is analysed by the second separator. Chromatographic resolution was not lost during the transfer process from the first to the second separation stages. The interface also allows for the exchange of the carrier gas used in the second gas chromatographic separation to provide for the maximum separation speed. In the first separation, a silica gel packed column and the novel application of a silica gel porous layer open tubular capillary column was used for SFC group separation. The SFCxGCftp was applied to petrochemical samples and essential oils and the results were compared to that obtained with a commercially available GCxGC system. / Thesis (PhD)--University of Pretoria, 2003. / Chemistry / unrestricted