Thesis (Ph. D. (Pharmacy))--Rhodes University, 2006.

Thesis (M.S.)--West Virginia University, 2006. / Title from document title page. Document formatted into pages; contains vii, 60 p. : ill. Includes abstract. Includes bibliographical references.

Thesis (Ph. D.)--University of Hong Kong, 2006. / Also available online.

Thesis (M.S.) University of Missouri-Columbia, 2006. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on August 24, 2007) Includes bibliographical references.

Thesis (M.S.)--Baylor University, 2009. / Includes bibliographical references (p. 83-88).

Thesis (MSc Food Sc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Phenolic compounds of the types present in plums have been found to exhibit health-promoting properties associated with their antioxidant capacity. Fruits with red peel and/or flesh are thus sought-after for their high antioxidant levels. In the current study South African plum (Prunus salicina Lindl.) cultivars and selections, harvested during two consecutive fruit seasons, were compared in terms of general fruit attributes (colour, firmness, °Brix, pH, titratable acidity), phenolic composition and antioxidant capacity. The effect of season and a commercial cold storage and ripening regime was also investigated. A reversed-phase high-performance liquid chromatography-diode-array-fluorescence detection (HPLC-DAD-FLD) method suitable for use with mass spectrometry (MS) detection, was optimised for separation and identification of phenolic compounds from four phenolic groups (phenolic acids, anthocyanins, flavan-3-ols and flavonols) in six South African plum cultivars and five selections. Parameters that were optimised include the mobile phases, analysis temperature and gradient program. Good stability, linearity and inter- and intra-day precision were obtained. Identification of compounds was based on comparison of retention times, UV-Vis spectra and mass fragments with available authentic phenolic standards and/or literature data. The optimised method allowed identification or tentative identification of twenty-four phenolic compounds, including cyanidin-3-O-glycosides, quercetin glycosides, monomeric, dimeric and trimeric flavan-3-ols, and hydroxycinnamic acids. An on-line ABTS•+ (2,2ʹ-azinobis-(3-ethyl-benzothiazoline-6-sulfonic acid)) antioxidant assay, performed for qualitative evaluation of the antioxidant response of individual phenolic compounds, indicated the flavan-3-ols as major antioxidants in plums. Eighteen phenolic compounds were quantified, including anthocyanins and flavonol glycosides, flavan-3-ols (monomers and dimers) and hydroxycinnamic acids. Phenolic composition differed greatly between cultivars and selections. Cyanidin-3-O-glucoside was the predominant anthocyanin in plums with red peel and/or flesh, followed by cyanidin-3-O-rutinoside. Cyanidin-3-O-galactoside was present only in the cultivar Laetitia (red peel, yellow flesh). The ripe fruit of Ruby Red and PR04-19, both with red peel and flesh, had the highest anthocyanin content for the first and second harvest season, respectively. Neochlorogenic acid and quercetin-3-O-glucoside were the major phenolic acid and flavonol, respectively. Chlorogenic acid, 3-O-p-coumaroylquinic acid and several quercetin-glycosides and -diglycosides were also present in some cultivars and selections. Procyanidin B1 was the flavan-3-ol present in the highest concentration in the majority of cultivars and selections and its content correlated with the (+)-catechin content, while the same was observed for procyanidin B2 and (-)-epicatechin. The effect of cold storage and ripening on fruit attributes differed greatly between cultivars and selections. The increase and decrease in pH and titratable acidity, respectively, were as expected for ripe fruit as opposed to unripe fruit. Ripe fruit had higher a*-values and lower L*-values. The cold storage and ripening regime had no significant effect on total polyphenol and total flavan-3-ol content of the cultivars and selections, but the anthocyanin content increased in some cases. In terms of in vitro antioxidant capacity, the selections PR04-32 and PR04-35, both with red peel and flesh, had the highest antioxidant capacity, irrespective of assay. Sapphire (red peel, yellow flesh), with the lowest total polyphenol content, also had the lowest antioxidant capacity in the ORAC and FRAP assays. Both the total polyphenol and flavan-3-ol contents correlated significantly to antioxidant capacity, irrespective of assay. / AFRIKAANSE OPSOMMING: Fenoliese verbindings, soos teenwoordig in pruime, is bekend vir hul gesondheidsbevorderende eienskappe wat geassosieer word met antioksidant kapasiteit. Vrugte met ‘n rooi skil en/of vleis is veral gesog as gevolg van hul hoë antioksidant aktiwiteit. Met hierdie studie is Suid-Afrikaanse pruim (Prunus salicina Lindl.) kultivars en seleksies, geoes tydens twee opeenvolgende seisoene, vergelyk in terme van algemene vrug einskappe (kleur, fermheid, °Brix, pH en titreerbare suurheid), fenoliese samestelling en antioksidant kapasiteit. Die effek van ‘n kommersiële koelopberging en rypwording prosedure is ook ondersoek. ‘n Omgekeerde-fase hoë-druk vloeistof chromatografie (HPLC) metode met diode-opstelling en fluoressensie deteksie, maar wat ook geskik is vir massa spektrometrie (MS), is geoptimiseer om fenoliese verbindings te skei en te identifiseer in Suid-Afrikaanse pruime. Verbindings van vier fenoliese groepe (fenoliese sure, antosianiene, flavan-3-ole en flavonole) wat in ses kultivars en vyf seleksies voorgekom het, is ondersoek. Die vloeistof fases, skeidingstemperatuur en gradiënt van die metode is geoptimiseer. Goeie resultate vir stabiliteit, lineariteit en inter- en intra-dag akkuraatheid is verkry. Verbindings is geïdentifiseer deur vergelyking van retensie tye, UV-Vis spektra en massa fragmente met dié van egte fenoliese standaarde en/of met literatuur data. Vier-en-twintig fenoliese verbindings is geïdentifiseer of voorlopig geïdentifiseer, insluitende sianidien- en kwersetien glikosiede, flavan-3-ol monomere, dimere en trimere, en hidroksikaneelsure. ‘n Aanlyn ABTS•+ (2,2 ʹ-azino-di-(3-etielbensotialosien-sulfoon suur) radikaal katioon blussingstoets is gebruik om die antioksidant reaksie van individuele polifenole op ‘n kwalitatiewe wyse te evalueer en flavan-3-ole is as hoof antioksidante in pruime aangetoon. Kwantifisering van agtien verbindings, insluitende antosianiene, flavonol glikosiede, flavan-3-ole (monomere en dimere) en hidroksikaneelsure, was moontlik met hierdie geoptimiseerde metode. Die fenoliese samestelling het aansienlik verskil tussen kultivars en seleksies. Sianidien-3-O-glukosied was die hoof antosianien in pruime met ‘n rooi skil en/of vleis, gevolg deur sianidien-3-O-rutinosied. Sianidien-3-O-galaktosied het slegs in Laetitia (rooi skil en geel vleis) voorgekom. Ryp vrugte van Ruby Red en PR04-19, beide met rooi skil en vleis, het onderskeidelik die hoogste antosianieninhoud gehad met die eerste en tweede seisoen se oeste. Neochlorogeniese suur en kwersetien-3-O-glukosied was die hoof fenoliese suur en flavon-3-ol, onderskeidelik. Chlorogeniese suur, 3-O-p-kumarienkwiniensuur en verskeie kwersetien glikosiede en diglikosiede was teenwoordig in sekere kultivars/seleksies. Die flavan-3-ol, prosianidien B1, was teenwoordig in die hoogste konsentrasie in die meerderheid kultivars/seleksies. Die prosianidien B1 inhoud het met die (+)-katekien inhoud gekorreleer, terwyl dieselfde gevind is vir prosianidien B2 en (-)-epikatekien. Die effek van koelopberging en rypwording op die algemene vrug einskappe het tussen kultivars en seleksies verskil. Die pH en titreerbare suurheid het onderskeidelik toegeneem en afgeneem, soos verwag is vir ryp vrugte teenoor onryp vrugte. Ryp vrugte het hoër a*-waardes en laer L*-waardes getoon. Koelopberging en rypwording het geen beduidende effek op die totale polifenol- en totale flavan-3-ol inhoud gehad nie, maar die antosianieninhoud het vir sommige kultivars en seleksies toegeneem. In terme van in vitro antioksidant kapasiteit het die seleksies PR04-32 en PR04-35, beide met rooi skil en vleis, die hoogste antioksidant kapasiteit getoon, ongeag die antioksidant toets wat gebruik is. Sapphire (rooi skil en geel vleis) het die laagste totale polifenolinhoud gehad, asook die laagste antioksidant kapasiteit soos bepaal deur die ORAC en FRAP toetse. Beide die totale polifenol- en flavan-3-ol inhoud het beduidend met die antioksidant kapasiteit korreleer, ongeag van die toets wat gebruik is.

Marine brown algae (seaweeds) are a rich source of fucoxanthin, a xanthophyll carotenoid that is naturally, an accessory pigment in the process of photosynthesis of sea vegetation such as Sargassum incisifolium. Fucoxanthin has been exploited by nutraceutical companies for its anti-obesity effects that has resulted in an increase of seaweed slimming preparations such as FucoThin™. The field is getting widespread consumer attention as interest in fucoxanthin has also transcended to its widespread biological potential which include cytotoxicity, anti-diabetic, anti-oxidant, anti-inflammatory and anti-plasmodium effects. We therefore wanted to identify a reliable source(s) of fucoxanthin from diverse samples of South African marine brown algae in order to explore our medicinal chemistry interests around the cytotoxicity and anti-malarial potential of fucoxanthin. A known source, Sargassum incisifolium, was used to isolate (maceration in CH₂Cl₂/MeOH at 35 °C followed by a hexane/EtOAc step gradient silica column of the crude extract and reversed phase semi-prep HPLC) and characterize (1D and 2D NMR) fucoxanthin (reference standard) in order to develop an analytical method for its determination in selected diverse brown algae commonly found in South Africa. The HPLC [Column: Phenomenex® Synergi™ (250 x 3.0 mm i.d); Mobile phase: ACN/H2O (95:5)] method developed for this analysis was validated according the guidelines set by the International Conference on Harmonization (ICH). Fifteen species were then assessed for fucoxanthin content (μg/g of dried weight) using the developed method. Stability studies on fucoxanthin were also carried out to assess photo- and pH degradation of fucoxanthin. Zonaria subarticulata (KOS130226-18) from Kenton-On-Sea beach and Sargassum incisifolium (PA130427-1) from Port Alfred beach were found to be the highest producers of fucoxanthin with 0.50 mg/g and 0.45 mg/g dried weight respectively. Fucoxanthin was found to be both photo-labile and sensitive to both acidic and basic pH environments. However, the pigment was more photostable in pure as opposed to extract form and also showed to be more stable at pH 10.0. Our findings show that Z. subarticulata and S. incisifolium could be reliable sources of fucoxanthin and can be considered as the algae to use in optimized extraction procedures in further studies. Also, when working with fucoxanthin, it is important to protect it from light. Any consideration of taking fucoxanthin preparation orally (as a nutraceutical) should consider protecting the active from the harsh conditions of the gastrointestinal tract. Any upscale production of fucoxanthin from seaweed should consider variations such as geographical, seasonal, lifecycle stage, etc. of identified algae as these may be important factors in obtaining effective concentrations of fucoxanthin.

A análise de impurezas é uma etapa importante no controle de qualidade do insumo farmacêutico e do produto final. Provenientes da síntese do medicamento ou dos excipientes, mesmo em pequenas concentrações, as impurezas podem afetar a eficácia e a segurança. No presente trabalho foram desenvolvidos e validados dois métodos empregando cromatografia líquida de alta eficiência (CLAE) para a avaliação do besifloxacino e sua impureza de síntese, um com eluição isocrática e outro com eluição gradiente. As análises por CLAE com modo de eluição isocrática foram executadas utilizando coluna Ciano, mantida a 25 °C. A fase móvel foi constituída por 0,5% de trietilamina (pH 3,0) : acetonitrila (88:12 v/v), eluída na vazão de 1,0 mL/min com detecção a 330 nm. O método com eluição gradiente foi conduzido com a mesma coluna e componentes da fase móvel modificando apenas as proporções entre fase orgânica e aquosa durante as análises. Os procedimentos foram validados de acordo com guias aceitos internacionalmente, observando-se resultados dentro dos limites aceitáveis. Os métodos apresentaram-se lineares na faixa de 140 a 260 μg/mL para o besifloxacino e de 0,3 a 2,3 μg/mL para a impureza A. Com volume de injeção de 20 μL, os limites de detecção e quantificação foram, respectivamente, 0,07 e 0,3 μg/mL. A precisão foi alcançada para todas as análises realizadas, obtendo DPR inter-dia igual a 6,47 e 6,36 para a impureza A com eluição isocrática e gradiente, respectivamente. A exatidão foi superior a 99% e a robustez apresentou resultados satisfatórios. No método isocrático obteve-se tempo de análise de 25 min e no gradiente de 15 min. O número de pratos teóricos no modo isocrático foi na ordem de 5000 enquanto no modo gradiente foi na ordem de 45000, ou seja, obteve-se maior eficiência da coluna com alteração da composição da fase móvel durante a eluição. No insumo besifloxacino e no produto farmacêutico utilizados neste trabalho, impurezas relacionadas estavam presentes, mas a impureza A não foi detectada. Os métodos propostos, considerando-se o limite de quantificação, podem ser aplicados na determinação quantitativa da impureza A na análise da matéria prima do besifloxacino, assim como na suspensão oftálmica. / Analysis of impurities is an important step in quality control of pharmaceutical ingredients and final products. From drug synthesis or excipients, even in small concentrations, impurities may affect efficacy and safety. In the present study two chromatographic methods were developed and validated for high-performance liquid chromatography (HPLC) for the assessment of besifloxacin and its synthesis impurity, one with isocratic and another with gradient elution. Analyses by HPLC in isocratic elution mode were performed using Cyano column maintained at 25 °C. Mobile phase was composed of 0.5% triethylamine (pH 3.0): acetonitrile (88:12 v/v) eluted at a flow rate of 1.0 ml/min with detection at 330 nm. The method with gradient elution was carried out with the same column and mobile phase components modifying only proportion between organic and aqueous phase during analysis. The procedures have been validated according to internationally accepted guidelines, obtaining results within acceptable limits. The methods presented a linear response from 140-260 μg/mL for besifloxacin and from 0.3 to 2.3 mg/mL for impurity. With the injection volume of 20 μL, the limit od detection and limit of quantitation were, respectively, 0.07 and 0.3 μg/mL. Precision was achieved for all analyses, obtaining inter-day RSD equal to 6.47 and 6.36 for impurity A with isocratic and gradient elution, respectively. The accuracy was higher than 99% and robustness exhibited satisfactory results. In the isocratic method was obtained analysis time 25 min and 15 min gradient. The number of theoretical plates in the isocratic mode was of the order of 5000 while in the gradient mode was of the order of 45000, that is, gave greater efficiency of the column by changing the mobile phase composition during elution. In raw material of besifloxacin and pharmaceutical product used in this study, related impurities were present but the impurity A was not detected. The proposed methods, considering the limit of quantitation, can be in quantitative determination of impurity A in the analysis of besifloxacin raw material, as well as in ophthalmic suspension.

A vigabatrina (VGB) é um fármaco anticonvulsivante que apresenta apenas a forma farmacêutica sólida disponível para uso. Na área hospitalar, devido à ausência de medicamentos na forma farmacêutica líquida, são preparadas derivações a partir de comprimidos e cápsulas para adequar a administração da dose prescrita. No entanto, a falta de estudos de estabilidade podem comprometer a eficácia e segurança destas derivações. O objetivo deste trabalho foi analisar a estabilidade química de derivações de comprimidos de vigabatrina em condições de armazenamento sob diferentes temperaturas e variações na embalagem utilizada. A análise das derivações de VGB foi realizada através de cromatografia líquida de alta eficiência (CLAE). O método descrito na Farmacopeia Britânica (2016) foi covalidado quanto a especificidade, linearidade, precisão e exatidão. Amostras de derivação de VGB foram preparadas em triplicata e acondicionadas em frascos de vidro e de PET âmbar. Após, foram armazenadas sob três diferentes condições de temperatura: temperatura ambiente (15 a 30 °C), sob refrigeração (2 a 8 °C) e em estufa (40 °C). Foram coletadas amostras armazenadas nas diferentes embalagens a cada 7 dias, por um período de 35 dias para as amostras conservadas em temperatura ambiente e refrigerada. O mesmo procedimento foi realizado para as amostras conservadas em estufa, porém por um período de 28 dias Também foi analisado o pH das amostras em cada tempo de coleta. As derivações de VGB foram analisadas por CLAE e apresentaram variação dentro dos limites preconizados pela Farmacopeia Britânica 2016, até 21 dias para frascos de vidro e de PET âmbar para as temperaturas ambiente e refrigerada. As amostras de VGB conservadas em estufa, apresentaram redução acima de 10% após 7 dias de estudo. A menor variação de pH ocorreu em frasco de vidro âmbar armazenado sob refrigeração. O resultado deste estudo serve de referência no preparo de derivações de VGB para uso hospitalar, pois apresentou intervalo de tempo confiável e condições de armazenamento adequadas para sua utilização. Desta forma, os pacientes pediátricos que utilizam doses fracionadas ou pacientes em uso de sondas nasogástricas terão as derivações adequadamente preparadas, reduzindo o risco de erros de diluição e contaminação microbiológica, melhorando a eficácia e segurança terapêutica. / Vigabatrin (VGB) is an anticonvulsant drug that has only solid dosage form available for use. In hospital, due to lack of medicines in liquid dosage form, extemporaneous preparations are prepared from tablets and capsules to adapt the administration of the prescribed dose. However, the lack of stability studies may compromise the efficacy and safety of these preparations. The aim of this study was to analyze the chemical stability of VGB extemporaneous preparation from tablets at storage conditions in different temperatures and variations of packaging used. The analysis of VGB extemporaneous preparations were performed by high-performance liquid chromatography (HPLC). The method described in British Pharmacopoeia (2016) was co-validated for specificity, linearity, precision and accuracy. VGB extemporaneous preparations were prepared in triplicate and placed in amber glass and PET bottles, which were stored under three different conditions: at room temperature (15 to 30 °C), under refrigeration (2 to 8 °C), and oven (40 °C). Samples of preparations stored at room temperature and refrigeration were collected every 7 days along 35 days. The same was done for solutions kept at 40 °C, but for a period of 28 days. It was also analyzed the preparations pH for each sampling time VGB extemporaneous preparations were analyzed by HPLC and demonstrated variations within the limits of British Pharmacopoeia (2016) up to 21 days in amber glass and PET bottles at room and refrigerated temperatures. VGB content for preparations kept in oven decreased above 10% after 7 days of study. The lowest pH change occurred in amber glass bottle stored under refrigeration. Results of this study can be applied as a reference for VGB extemporaneous preparation in hospital, once it was demonstrated the reliability of storage time interval and proper conditions for the use. Thus, pediatric patients with fractionated doses or patients using nasogastric probe will have adequately prepared extemporaneous preparations, reducing the risk of dilution errors and microbiological contamination.

Made available in DSpace on 2014-06-11T19:25:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-02Bitstream added on 2014-06-13T20:33:02Z : No. of bitstreams: 1 cazedey_ecl_me_arafcf.pdf: 802368 bytes, checksum: 7af4d6526e6d1c491bb231f706185aea (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O cloridrato de ciprofloxacino, um antibacteriano quinolônico, apresenta amplo espectro de ação e é eficaz in vitro contra praticamente todos os patógenos Gram-negativos, incluindo Pseudomonas aeruginosa. Mostra-se também eficaz contra microrganismos Gram-positivos, como estafilococos e estreptococos. A importância de desenvolver e validar métodos analíticos para este fármaco é justificada por seu potencial terapêutico, grande emprego em terapias microbianas e baixo custo, assim como pelo conhecimento de que a baixa qualidade dos produtos anti-infecciosos está relacionada ao desenvolvimento de cepas resistentes, como consequência da administração de doses subterapêuticas. Por tal razão, é de enorme importância o desenvolvimento de métodos analíticos eficazes e confiáveis para o controle de qualidade dos medicamentos comercializados. Neste trabalho foram desenvolvidos métodos de análise para o cloridrato de ciprofloxacino em solução oftálmica. Os métodos desenvolvidos e validados foram: (i) doseamento microbiológico, método turbidimétrico na faixa de concentração de 14,0 a 56,0 μg/mL, utilizando Staphylococcus epidermidis ATCC 12228 IAL 2150, com exatidão de 99,71% e teor de 102,27%; (ii) método espectrofotométrico na região do UV a 275 nm com faixa de concentração de 2,0 a 7,0 μg/mL, utilizando água como solvente, com exatidão de 101,51% e teor de 99,79%; (iii) método espectrofotométrico derivativo na região do visível a 386,4 nm, na primeira derivada, com faixa de concentração de 50,0 a 100,0 μg/mL, utilizando cloreto férrico 1,0% como reagente, com exatidão de 99,83% e teor de 106,72%; (iv) método por cromatografia líquida de alta eficiência com detector UV a 275 nm, com fase móvel composta por ácido acético 2,5% v/v, metanol e acetonitrila (70:15:15, v/v/v) e faixa de concentração de 1,0 a 6,0 μg/mL, exatidão... / Ciprofloxacin hydrochloride, a quinolone antibiotic, presents a wider spectrum of activity and is effective against practically all Gram-negative pathogens, including Pseudomonas aeruginosa. It is potent against Grampositive microorganisms, as Staphylococcus and Streptococcus. Analytical methods for quantitative determination of ciprofloxacin hydrochloride is important due to its therapeutic potential, wide use in antimicrobial therapy and low cost. Moreover, it is known that the poor quality antibiotic product is direct related development of resistant strains, as consequence of subtherapeutic doses administration. Thus, it is important to develop efficient analytical methods for quality control commercialized products. In this work, analytical methods for determination of ciprofloxacin hydrochloride were validated: (i) microbiological assay, turbidimetric method at concentration range 14.0 to 56.0 μg/mL, using Staphylococcus epidermidis ATCC 12228 IAL 2150 as indicator microorganism, accuracy 99.71% and quantitation of 102.27; (ii) UV spectrophotometry at 275 nm with concentration range of 2.0 to 7.0 μg/mL, using water as solvent, with accuracy of 101.51% and quantitation of 99.79%; (iii) Derivative visible spectrophotometric method at 386.4 nm, in first derivate, with concentration range of 50.0 a 100.0 μg/mL, using 1.0% ferric chloride as reagent, with accuracy of 99.83% and quantitation of 106.72%; (iv) HPLC method with UV detector at 275 nm using 2.5 M acetic acid (v/v), methanol and acetonitrile (70: 15: 15, v/v/v) as mobile phase and concentration range of 1.0 to 6.0 μg/mL, accuracy of 100.11%, quantitation 103.25% and mean retention time of 2.6 minutes; (v) Indirect titrimetric method using bromate/bromide solution in acid medium as reagent in concentration range of 1.0 to 11.0 mg/mL, with accuracy of 100.28% and quantitation of 98.97%.