Thesis (Ph. D.)--University of Hong Kong, 1973. / Typewritten.

published_or_final_version / Pathology / Doctoral / Doctor of Philosophy

In this diploma thesis, capillary columns with an inner diameter of 530 microns filled with gel particles were prepared. These columns are intended for gel chromatography. Three various methods of filling were used for their preparation. The first method was based on filling of wet particles in suspension with the organic solvent, followed by drying and swelling of particles with water into the form of gel. The second method used filling of wet particles in the suspension form with the aqueous phase. The last method started with suction of dry gel particles by air and continued with their subsequent swelling with water into the form of the gel. The prepared columns have been compared analysing hyaluronic acid and thiourea. The basic parameters specified for the gel column chromatography were determined by using phenylalanine. The pressure characteristics depending on the flow rate of mobile phase were used to compare the quality of packing of the columns prepared by various methods.

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Submitted by Amanda Silva (amanda.osilva2@ufpe.br) on 2015-04-08T12:27:25Z No. of bitstreams: 2 Dissertação Mestrado Final.pdf Rosimery.pdf: 719273 bytes, checksum: b4f8824614b283e70abb1c89cc2001f2 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-04-08T12:27:25Z (GMT). No. of bitstreams: 2 Dissertação Mestrado Final.pdf Rosimery.pdf: 719273 bytes, checksum: b4f8824614b283e70abb1c89cc2001f2 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2012 / CNPq / Hemoderivados são medicamentos produzidos pelo fracionamento industrial do plasma humano. O principal método para purificação de proteínas plasmáticas humanas é baseado no método de precipitação com etanol desenvolvido por Cohn-Oncley. O uso de metodologias simples, de baixo custo e eficientes constitui um avanço tecnológico para obtenção desses hemoderivados. O objetivo deste trabalho foi imobilizar heparina comercial em Sephadex G-25 revestido com polianilina e posteriormente utilizar o derivado imobilizado como matriz de afinidade para purificação de proteínas do plasma humano. Para síntese do derivado imobilizado, Sephadex G-25 foi revestido com polianilina e tratado com glutaraldeído, em seguida, incubado com solução de heparina ativada com 1-etil-3-(dimetilaminopropil) carbodiimida e N-hidroxisuccinimida. A fim de determinar a influencia de variáveis na imobilização de heparina em Sephadex-polianilina, foi realizado planejamento experimental fatorial fracionário (24-1) no qual se avaliou quatro variáveis independentes: concentração e tempo de reação do glutaraldeído e concentração e tempo de reação da heparina. Nas condições otimizadas desses níveis, a heparina imobilizada em Sephadex-polianilina foi utilizada para purificação de proteínas do plasma humano. As variáveis concentração de glutaraldeído e concentração de heparina foram estatisticamente significativas na imobilização de heparina ao Sephadex-PANI e foi obtido melhor rendimento de heparina imobilizada (6,88 μg mg-1 suporte, equivalente a 45,9% da heparina ofertada) nas condições 1% (v v-1) de glutaraldeído, 1,5 h de reação com glutaraldeído, 1,5 mg mL-1 de concentração de heparina e 2 h de reação com heparina. Nessas condições obteve-se uma quantidade de proteínas plasmáticas de 25,02 – 27,06 μg mL-1, nos eluatos de NaCl 0,5 a 2,0 mol L-1. A presença de proteínas foi confirmada através da técnica eletroforética em gel de poliacrilamida (SDS-PAGE). O compósito sephadex-polianilina-heparina foi eficiente para purificação de proteínas do plasma, podendo ser usado como teste a antitrombina.

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Triglycerides are the majority lipid component of most biochemical mixtures and are virtually water insoluble. Lipid removal is desired prior to protein purification processing to decrease nonspecific fouling of downstream chromatographic matrices. Transgenic pig milk was used as a model system to study delipidation from therapeutic protein sources. The majority of triglycerides was extracted from stable lipid micelles and removed with a method that can be incorporated in downstream protein purification processing without denaturing the target protein. An efficient delipidation treatment used TNBP, a non-polar solvent, to extract lipid micelles and then phase transfer milk lipids into a TNBP-swelled dextran particulate. A batch incubation of a whey/TNBP mixture with pre-swollen Sephadex LH-20 or hydroxyalkoxypropyl dextran (HAPD) beads at 4 C for 24 hours removed 67 + 2 % (0.645 mg triglycerides/ml Sephadex LH-20) and 71 o + 1 % (0.628 mg triglycerides/ml HAPD) of the triglycerides present in the skimmed transgenic whey, respectively. Fully swollen beads removed 20% more triglycerides than beads which were wetted but not swollen in TNBP, indicating that a larger phase volume and internal adsorption of the lipids onto the Sephadex matrix dominates over surface adsorption. Polyclonal ELISAs indicated that 89 + 6% of the recombinant human Protein C was still present in the transgenic whey after this delipidation treatment, indicating this treatment did not denature or harm the target protein. / Master of Science

Medicinal plants which have been widely used in folk medicine are known to contain important biologically active compounds. Most of today&rsquo / s synthetic drug raw materials are to be prepared by using plant originated compounds as the starting material. Hypericum Perforatum is one of the medicinal plants that grows in Europe, Western Asia and Northern Africa and is distinguished by its golden yellow flowers. The common name of the plant is St. John&rsquo / s wort. From the time of the ancient Greeks down through the middle Ages, the plant was considered to be imbued with magical powers and was used to ward off evil and protect against disease. As a practical folk-remedy, it has been used widely to heal wounds, remedy kidney troubles, and alleviate nervous disorders, even insanity. In the last thirty years, Hypericum perforatum has undergone extensive clinical and laboratory testing. The extract of the flower is a red liquid that contains many biologically active compounds such as: naphtodianthrones (hypericin, pseudohypericin), phloroglucinols (hyperforin, adhyperforin), flavonoids (quercetin, hyperoside, quercitrin, isoquercitrin, rutin, campferol, myricetin, amentofloavone), procyanidins (procyanidin, catechin, epicatechin polymers) , tannins (tannic acid), essential oils (terpenes, alcohols), amino acids (GABA, Cysteine, glutamine, leucine, lysine, ornithine, praline, threonine), phenyl propanes (caffeic acid, chlorogenic acid), xanthones (keilcorin, norathriol), organic acids peptides and polysaccharides (other water soluble compounds). These compounds have previously been isolated using HPLC method. The aim of this study is to isolate the main biologically active compound groups of Hypericum Perforatum and simply characterize the compounds with TLC, UV-VIS spectroscopy, NMR spectroscopy using standard compounds as references.

Magister Scientiae - MSc (Chemistry) / Plants have been widely used in traditional medicine for a number of ailments, among which may be included infectious diseases such as colds, influenza, chicken pox, TB, etc. as well as lifestyle diseases such as diabetes and cancer. Seaweeds have also been shown to contribute to the maintenance of health through their nutritional and medicinal properties and recently, a great deal of interest has developed towards the isolation of bioactive compounds from marine sources due to their numerous health benefits. Furthermore, marine algae are valuable sources of structurally diverse metabolites with scientifically proven therapeutic claims. Chemical constituents of red seaweed, Porphyra capensis was investigated in this present study along with subsequent brine shrimp lethality assay analysis of the crude extracts. The compounds isolated from the plant were from the hexane (6) and butanol (2) extracts. These compounds were all isolated and purified by various chromatographic techniques, namely silica gel chromatography, Sephadex LH-20 gel as well as C18 reversed phase silica gel. The structures of the isolated compounds were analysed and characterised by NMR, GC-MS, ESI MS and FTIR spectroscopy. Eight compounds were isolated and identified as phytol, desmosterol, 9-eicosenoic acid, 5,8,11,14,17-eicosapentanoic acid, palmitic acid, methyl (E)-hexadec-9-enoate, glycerol and compound 1 (novel compound). All the compounds were isolated from Porphyra capensis for the first time. The hexane, butanol and methanol extracts were found to be non-toxic with the brine shrimp test LC50 value at least two times greater than ?g/ml.