A research report Submitted to the faculty of Health Sciences, University of Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the degree of Masters of Science in Medicine (Pharmaceutical Affairs). / Pathogenic mycobacterial infection was in recent decades a health concern so well controlled that eradication seemed imminent. However, it is once again reaching epidemic proportions following the increasing prevalence of AIDS. One important means of curbing this resurgence, is a robust method that has the capability of identifying to a species level speciating disease causing mycobacteria in a matter of days. Several new methodologies are now available that enable dramatic reductions in turn-around times. In this study High Performance Liquid Chromatography was investigated to determine how this system compared with the current mycobacterial system of methodologies adopted in South African laboratories. Four species of pathogenic mycobacteria, with a high prevalence in South Africa, were tested in a sample size of 80. Samples were subjected to HPLC, Gene Probes and Biochemical testing. HPLC was the most capable of identifying the mycobacteria to species level displaying a sensitivity to the organisms of 96.25 %. Gene probes and Biochemical testing had sensitivity values of 82.5 % and 80 % respectively. HPLC was also more cost efficient and displayed a wider range of identification. It is therefore suggested that HPLC replace Gene probes and Biochemical testing for purposes of MOTT identification in the comprehensive mycobacterial identification system. The result is a time saving of at least 3 weeks and a cost reduction of approximately 30 %. The large initial capital investment required for the implementation of the HPLC system is justified by the long term cost saving as well as the additional utility derived from early identification. As a consequence treatment is not empiric but rather tailored to the organism infecting the patient, hence preventing multiple drug resistance developing and ultimately saving a life through rational drug use. / WHSLYP2016

by Lau Yick Ki. / Thesis (M.Ph.)--Chinese University of Hong Kong, 1987. / Includes bibliographies.

Poon Wai Mei Emily. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 74-81). / Abstracts in English and Chinese. / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- Importance of amino acids --- p.1 / Chapter 1.1.1. --- Clinical samples --- p.1 / Chapter 1.1.2. --- Food samples --- p.2 / Chapter 1.2. --- Reviews of amino acid analysis --- p.6 / Chapter 1.2.1. --- Ion-exchange chromatography --- p.6 / Chapter 1.2.2. --- Gas chromatography --- p.6 / Chapter 1.2.3. --- Thin layer chromatography --- p.8 / Chapter 1.2.4. --- Flow injection analysis --- p.8 / Chapter 1.2.5. --- Liquid chromatography --- p.9 / Chapter 1.2.6. --- Capillary electrophoresis --- p.10 / Chapter 1.2.7. --- Methods of detecting amino acid without derivatization --- p.11 / Chapter 1.3. --- Determination of amino acids by reversed-phase ion-pair chromatography --- p.12 / Chapter 1.4. --- The objectives of the study --- p.15 / Chapter Chapter 2: --- Experimental --- p.16 / Chapter 2.1. --- Materials --- p.16 / Chapter 2.2. --- Apparatus --- p.16 / Chapter 2.3. --- Samples --- p.16 / Chapter 2.4. --- Procedures --- p.17 / Chapter 2.4.1. --- Preparation of amino acid standard solution (stock solutions) --- p.17 / Chapter 2.4.2. --- Method development --- p.17 / Chapter 2.4.3. --- Samples preparation --- p.18 / Chapter 2.4.4. --- Preparation of Dowex column --- p.18 / Chapter 2.4.5. --- Extraction of amino acids from samples --- p.19 / Chapter 2.4.6. --- Recovery test --- p.20 / Chapter Chapter 3: --- Results and Discussions --- p.21 / Chapter 3.1. --- Optimization --- p.21 / Chapter 3.1.1. --- pH --- p.21 / Chapter 3.1.2. --- Ion-interacting reagent --- p.22 / Chapter 3.1.3. --- Organic solvent --- p.29 / Chapter 3.1.4. --- Temperature --- p.34 / Chapter 3.1.5. --- Chromatographic conditions --- p.36 / Chapter 3.2. --- Application --- p.45 / Chapter 3.2.1. --- Precision of injection --- p.45 / Chapter 3.2.2. --- Accuracy of the method --- p.46 / Chapter 3.2.3. --- The concentration of amino acids in food samples --- p.50 / Chapter 3.2.3.1. --- Citrus fruits --- p.50 / Chapter 3.2.3.2. --- Orange juice drinks --- p.60 / Chapter 3.2.3.3. --- Chinese honey --- p.65 / Chapter 3.2.3.4. --- New Zealand honey --- p.67 / Chapter 3.2.3.5. --- Energy drinks --- p.70 / Chapter Chapter 4 : --- Conclusion --- p.72 / Chapter Chapter 5 : --- Bibliographies --- p.74 / Chapter 6. --- Appendices --- p.82 / Chapter 6.1. --- Table 1 : Ingredients of orange juice drinks --- p.82 / Chapter 6.2. --- Table 2 : Honey samples --- p.83 / Chapter 6.3. --- Table 3 : Ingredients of energy drinks --- p.83

by Kwok Chiu Nga. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 1-3 (4th gp.)). / Abstract also in Chinese. / TABLE OF CONTENTS --- p.i / ABSTRACT --- p.iv / 摘要 --- p.vi / LIST OF FIGURES --- p.vii / LIST OF TABLES --- p.x / ABBREVIATION --- p.xi / Chapter CHAPTER ONE --- RESEARCH BACKGROUND / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.1.1 --- Alkaloids --- p.1 / Chapter 1.1.2 --- Diterpenoid alkaloids --- p.2 / Chapter 1.1.3 --- Aconitine-type alkaloids --- p.2 / Chapter 1.1.4 --- Toxicity --- p.4 / Chapter 1.1.5 --- Safety concerns --- p.4 / Chapter 1.2 --- Summary of the Previous Work --- p.8 / Chapter 1.3 --- Objectives and Outline of the Present Work --- p.13 / Chapter CHAPTER TWO --- INSTRUMENTATION AND EXPERIMENTAL / Chapter 2.1 --- Instrumentation --- p.15 / Chapter 2.1.1 --- High performance liquid chromatography (HPLC) --- p.15 / Chapter 2.1.2 --- Triple-stage quadrupole (TSQ) mass spectrometer --- p.17 / Chapter 2.1.2.1 --- Atmospheric pressure chemical ionization (APCI) --- p.17 / Chapter 2.1.2.2 --- Electrospray ionization (ESI) --- p.20 / Chapter 2.1.2.3 --- Quadrupole system --- p.20 / Chapter 2.1.2.4 --- Ion detection system --- p.22 / Chapter 2.1.2.5 --- Data system --- p.22 / Chapter 2.2 --- Experimental --- p.22 / Chapter 2.2.1 --- Sample and reagents --- p.22 / Chapter 2.2.2 --- Sample preparation --- p.23 / Chapter 2.2.3 --- High performance liquid chromatography conditions --- p.23 / Chapter 2.2.4 --- Mass spectrometry conditions --- p.25 / Chapter 2.2.4.1 --- Atmospheric pressure chemical ionization conditions --- p.25 / Chapter 2.2.4.2 --- Electrospray ionization conditions --- p.25 / Chapter CHAPTER THREE --- SELECTION AND OPTIMIZATION OF HPLC/MS METHOD / Chapter 3.1 --- Introduction --- p.26 / Chapter 3.2 --- Experimental --- p.29 / Chapter 3.3 --- Results and Discussion --- p.29 / Chapter 3.3.1 --- Triethylamine concentration --- p.31 / Chapter 3.3.2 --- Ammonium acetate concentration --- p.34 / Chapter 3.3.3 --- Acetic acid concentration --- p.37 / Chapter 3.3.4 --- HPLC/MS interface --- p.40 / Chapter 3.3.5 --- MS/MS conditions --- p.40 / Chapter 3.4 --- Conclusions --- p.43 / Chapter CHAPTER FOUR --- DETERMINATION OF ACONITINE-TYPE ALKALOIDS IN ACONITE ROOTS / Chapter 4.1 --- Introduction --- p.48 / Chapter 4.2 --- Experimental --- p.48 / Chapter 4.3 --- Results and Discussion --- p.50 / Chapter 4.3.1 --- Selection of internal standard --- p.50 / Chapter 4.3.2 --- Method validation --- p.50 / Chapter 4.3.2.1 --- Precision of measurement --- p.50 / Chapter 4.3.2.2 --- Accuracy of measurement --- p.50 / Chapter 4.3.2.3 --- Limits of detection and quantitation --- p.58 / Chapter 4.3.3 --- Determination of aconitine-type alkaloids in aconite roots --- p.58 / Chapter 4.4 --- Conclusions --- p.60 / Chapter CHAPTER FIVE --- CONCLUSIONS AND FUTURE WORK / Chapter 5.1 --- Conclusions --- p.67 / Chapter 5.2 --- Future Work --- p.68 / ACKNOWLEDGMENT --- p.A1 / APPENDIX --- p.A2 / REFERENCES --- p.R1

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 4 uppsatser.

An objective analytical method was developed to characterize the taste profiles of five cheese varieties. Nonvolatile water extracts of Cheddar, Edam, Gouda, Swiss, and Parmesan cheeses were analyzed by high performance liquid chromatography (HPLC) with a reversed phase column. The HPLC operating conditions were determined with Mapping Super-Simplex followed by Centroid Mapping Optimization. A ternary gradient elution system was used with an Adsorbosphere C8 column to resolve a maximum number of components. The optimum solvent volume ratio was 96.8 : 1.2 : 2.0 for trifluoroacetic acid (0.1%), acetonitrile, and methanol, with a flow rate of 1.0 mL/min. Over 50.3 min this ratio was changed to 56.3 : 30.3 : 13.4. Multivariate statistical analyses including principal component and discriminant analyses were applied to 55 peak areas from 106 cheese chromatograms. Principal component analysis reduced the dimensionality of the "data from 55 to 17 principal components, which are-combinations of the original variables, with a 26% loss of explained sample variation. Discriminant analysis on data from a single HPLC column was able to correctly classify cheeses by variety at a greater than 90% success rate. This grouping rate dropped to 64% when data from all four HPLC columns was combined, implicating large between column variations. A semi-trained sensory panel correctly classified cheeses by variety at a 63% rate. This objective method provides a lasting fingerprint of cheese products. / Land and Food Systems, Faculty of / Graduate

High Pressure Liquid Chromatography (HPLC) is a well-known and widely used analytical technique which is prevalent throughout the pharmaceutical industry as a research tool. Despite its prominence HPLC possesses some disadvantages, most notably slow analysis time and large consumption of organic solvents. Ultra Pressure Liquid Chromatography (UPLC) is a relatively new technique which offers the same separation capabilities of HPLC with the added benefits of reduced run time and lower solvent consumption. One of the key developments which facilitate the new UPLC technology is sub 2-µm particles used as column packing material. These particles allow for higher operating pressures and increased flow rates while still providing strong separation. Although UPLC technology has been available since early 2000, few laboratories have embraced the new technology as an alternative to HPLC. Besides the resistance to investing in new capital, another major roadblock is converting existing HPLC methodology to UPLC without disruption. This research provides a framework for converting existing HPLC methods to UPLC. An existing HPLC method for analysis of Galantamine hydrobromide was converted to UPLC and validated according to ICH guidelines. A series of statistical evaluations on the validation data were performed to prove the equivalency between the original HPLC and the new UPLC method. This research presents this novel statistical strategy which can be applied to any two methodologies to determine parity. / Chemistry

A Dissertation Submitted to the Faculty of Medicine, University of the Witwatersrand, Johannesburg, in Partial Fulfilment of the Requirements for the Degree of Master of Pharmacy Johannesburg, March 1994 / Preformulation and formulation study of dexchlor- pheniramine maleate (DCPM) for it's inclusion into a gelforming sustained release dosage form was investigated. A modification of the USP apparatus 2 is proposed as an alternative to currently recommended USP dissolution apparatus for floating, gelforming drug delivery systems. In addition, the role of magnesium stearate and talc as dissolution retardants in controlled release matrix tablets is investigated, through application of a factorial design. / IT2018

A azatioprina (AZA) é um imunossupressor utilizado no tratamento de doenças autoimunes como lúpus eritematoso sistêmico, doença de Crohn, doença inflamatória intestinal e contra a rejeição em transplantes de órgãos sólidos. Após mais de 40 anos de uso a AZA continua exercendo um papel central nos regimes imunomoduladores, devido ao fato de combinar eficácia, segurança e baixo custo. Sabe-se que a atividade da tiopurina metiltransferase pode determinar, pelo menos em parte, a eficácia clínica da AZA. Esta enzima apresenta polimorfismo genético co-dominante e a distribuição dos alelos variantes é significativamente diferente entre as populações. A grande variabilidade farmacocinética no metabolismo AZA justifica a sua monitorização terapêutica. Neste trabalho otimizou-se uma metodologia para a quantificação dos metabólitos da AZA, 6-TGN e 6-MMP, por cromatografia líquida de alta eficiência (HPLC/UV-Vis), utilizando-se um detector de ultravioleta-visível em um único comprimento de onda, após a amostra passar por uma desproteinização ácida simples e ser aquecida para a conversão dos metabólitos em suas respectivas bases livres. Os valores destes metabólitos foram determinados em uma população de 124 pacientes transplantados renais. Para adequarmos o processo às legislações locais e internacionais, foram seguidas orientações da Anvisa, FDA e CLSI. A separação foi realizada em coluna de fase reversa, sendo a fase móvel A fosfato de potássio e a fase móvel B metanol. A detecção da 6-TGN e da 6-MMP foi realizada em 342 m (UV-Vis). O estudo da linearidade da 6-TGN variou entre 0,30 e 89,71 mol/L e da 6-MMP entre 0,30 e 93,86 mol/L. As recuperações, de 95,08 a 100,80% para 6-TGN e 95,38 a 105,06% para 6-MMP. Os CV da repetibilidade, de 0,04 a 5,06%, enquanto os CV da reprodutibilidade de 4,88 a 12,73% para 6-TGN e 6-MMP. Para ambos os metabólitos o LD e o LQ foram de 0,30 mol/L e 0,13 mol/L. Os eritrócitos lavados e as amostras tratadas, prontas para a injeção no HPLC, foram armazenadas abaixo de -5°C até a análise. Nesta temperatura estiveram estáveis durante 8 semanas e 1 dia, respectivamente. Os valores das concentrações de 6-TGN e 6-MMP encontrados nas amostras dos pacientes variaram entre não detectável a 1569 mol/8 x 108 RBC (mediana de 200,50) e não detectável a 113057 mol/8 x 108 RBC (mediana de 5166), respectivamente. As correlações entre os níveis de 6-TGN ou 6-MMP e as variáveis sexo, tempo pós-transplante, número de transplantes e dosagem de AZA (mg/kg) foram examinadas em diferentes grupos. O método proposto apresenta boa relação custo-benefício, é simples, preciso e rápido na determinação das concentrações intraeritrocitárias de 6-TGN e 6-MMP em pacientes sob terapia com AZA. O método validado permite que o laboratório forneça dados farmacocinéticos úteis e precisos para o ajuste do tratamento do paciente e pode ser facilmente adaptado para a análise rotineira destes metabólitos. Os resultados das amostras dos pacientes estão de acordo com os encontrados em outros estudos, atestando a utilidade dessa ferramenta analítica no acompanhamento dos pacientes / Azathioprine (AZA) is an immunosuppressant used in autoimmune pathologies like lupus erythematosus, Chrons disease, inflammatory bowel disease and against rejection in solid organs transplant. After more than 40 years of use, AZA continues exerting a central role in immunomodulatory regimens, due to the fact that it combines effectiveness, safety and low cost. It is well known that thiopurine methyltransferase activity may determine, at least in part, the clinical efficacy of AZA therapy. This enzyme exhibits codominant genetic polymorphism and the distribution of these variant alleles differs significantly among populations. The considerable pharmacokinetic variability in AZA metabolism justify the therapeutic drug monitoring of this drug. In this work a methodology was improved to quantify the metabolites of AZA, 6-TGN and 6-MMP, by high performance liquid chromatography (HPLC/UV-Vis) with an ultraviolet-visible detector, using a single wavelength reading, following a simple acid deproteinization and heating to convert the metabolites into their respective free bases. The values of these metabolites were determined in a population of 124 renal transplant recipients. To adequate the process to international and local legislation, Anvisa, FDA and CLSI guidelines were followed. Separation was achieved on a reversed-phase column; mobile phase A potassium phosphate and mobile phase B methanol. Detection of 6-TGN and 6-MMP was performed at 342 m (UV-Vis). Assay linearity for 6-TGN ranged from 0.30 to 89.71 mol/L and from 0.30 to 93.86 mol/L for 6-MMP. The recoveries were 95.08, 97.76 and 100.80% for 6-TGN and 104.79, 95.38 and 105.06% for 6-MMP. Repeatability CV were 3.50, 5.06, 1.09 and 0.04, 0.35, 1.58%, while reproducibility CV were 8.65, 7.18, 8.44 and 12.73, 6.40, 4.88% for 6-TGN and 6-MMP, respectively. LOQ and LOD of 6-TNG and 6-MMP were respectively 0.30 mol/L and 0.13 mol/L for both metabolites. The washed erythrocytes and the samples treated and ready for injection into the HPLC system were stored below -5 °C until analysis, at this temperature the samples were stable for 8 weeks and for 1 day, respectively. 6-TGN and 6-MMP patient analysis values ranged from non detectable to 1569 mol/8 x 108 RBC (median of 200.50) and non detectable to 113057 mol/8 x 108 RBC (median of 5166), respectively. The correlations between 6-TGN or 6-MMP levels and variables sex, time post-transplant, number of transplants and AZA dosage (mg/kg) were examined in different groups. The proposed HPLC method has a good cost-benefit ratio, is straightforward, precise, accurate and fast at the determining 6-TGN and 6-MMP concentrations in red blood cells of patients under AZA therapy. The validated method is good enough to enable the laboratory to routinely provide useful and accurate pharmacokinetic data in time to adjust patient regimens. It can be easily adopted for routine analysis of these drug metabolites. The results of patient samples are in agreement with others studies, thus certifying the usefulness of this analytical tool in monitoring of patients

Desde 1964, o Brasil tem lançado programas de políticas públicas para melhorar o acesso da população aos medicamentos considerados essenciais. Em 1999, com a criação da Agência Nacional de Vigilância Sanitária e a introdução dos medicamentos genéricos no mercado brasileiro, o Brasil passou a ter três classes de medicamentos disponíveis no mercado farmacêutico: referência, similar e genérico. O objetivo deste estudo foi avaliar a bioequivalência e intercambialidade entre dois antipsicóticos (referência e similar) utilizados pelo Instituto de Psiquiatria do Hospital das Clínicas da Universidade de São Paulo, contendo 2 mg de risperidona. Foi desenvolvido e validado um método analítico que emprega a cromatografia líquida de alta eficiência acoplada à espectrometria de massas para a determinação da risperidona (RSP) e seu principal metabólito a 9-hidroxirisperidona (9OH-RSP) em plasma. Para se avaliar a bioequivalência entre os medicamentos foram recrutados 22 voluntários sadios, os quais participaram do estudo clínico conduzido de forma cruzada e aleatória. As coletas sanguíneas para o ensaio de bioequivalência foram realizadas em tubos heparinizados (5 mL) e os tempos de coleta foram 0 (antes da medicação); 0,25; 0,5; 1; 1,5; 3; 5; 8; 12; 24; 48; 72; 96 e 120 horas após a administração da medicação. A determinação da bioequivalência entre os dois medicamentos deu-se através da comparação dos parâmetros farmacocinéticos: concentração plasmática máxima (Cmax), tempo para atingir a concentração plasmática máxima (Tmax) e área sobre a curva de decaimento plasmático (ASCT). Os resultados obtidos foram submetidos à análise de variância (ANOVA) e foi adotado o intervalo de confiança de 90% (IC 90%). Os valores médios para Cmax, Tmax e ASCT para RSP para os medicamentos referência e teste foram 16,02 ng/mL; 1,5 h e 348,94 ng.h/mL e 12,65 ng/mL; 1,5 h e 286,03 ng.h/mL, respectivamente. Já os valores médios para Cmax, Tmax e ASCT para 9OH-RSP para os medicamentos referência e teste foram 21,00 ng/mL; 5,0 h e 821,40 ng.h/mL e 17,85 ng/mL; 5,0 h e 632,92 ng.h/mL. Os valores de IC 90% para Cmax e ASCT para RSP para os medicamentos referência e teste foram 74 a 82% e 76 a 85%, respectivamente, e os valores de IC 90% para os mesmos parâmetros para 9OH-RSP foram 83 a 87% e 75 a 78%, respectivamente. Os resultados demonstraram diferenças significativas entre os medicamentos testados, o que permite concluir que os mesmos não são bioequivalentes e, portanto, não podem ser intercambiáveis / Brazil has launched programmes of public policies aiming to improve essential medicines access for the population since 1964. It was created in 1999 the National Agency for Sanitary Vigilance, which introduced the generic medicines in the Brazilian market, which already had the reference and the pharmaceutical equivalent ones. The objective of this study was to evaluate the bioequivalence and interchangeability between two antipsychotics (reference and pharmaceutical equivalent) used by the Institute of Psychiatry, Hospital of the Universidade de São Paulo, containing 2 mg of risperidone. It was developed and validated a high-performance liquid chromatography coupled to mass spectrometry method for the determination in plasma of risperidone (RSP) and its main metabolite, 9- hydroxy-risperidone (9OH-RSP). To assess bioequivalence between the medicines it was recruited 22 healthy volunteers, which took part in a clinical cross and random studies. The blood collections were performed on heparinizades tubes (5 ml) and runtimes collections were 0 (before medication); 0.25; 0.5; 1; 1.5; 3; 5; 8; 12; 24; 48; 72; 96 and 120 hours after the administration of medication. The determination of bioequivalence between the two drugs was achieved by a comparison of the following pharmacokinetic parameters: plasma concentration (Cmax), time to achieve Cmax (Tmax), and area under the plasma concentration-time curve (AUCT). Results were subjected to analysis of variance (ANOVA), adopting a confidence interval CI 90%. The average values for Cmax, Tmax and AUCT for RSP were 16.02 ng/ml, 1.5 h and 348.94 ng.h/ml for reference medicines and 12.65 ng/ml, 1.5 h and 286.03 ng.h/ml for testing ones. The average values for Cmax, Tmax and AUCT for 9OH-RSP were 21.00 ng/ml, 5.0 h and 821.40 ng.h/ml for reference medicines and 17.85 ng/ml, 5.0 h and 632.92 ng.h/ml for testing ones. CI 90% for Cmax and AUC (RSP) were 74-82% and 76-85%, respectively. The CI 90% for the same parameters for 9OH-RSP was 83-87% for reference medicines and 75-78% for testing ones. There was significant difference between the products tested, thus one can conclude they are not bioequivalents, therefore cannot be interchanged