85 f. / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-04-08T13:22:40Z No. of bitstreams: 1 NORMANDO DISSERTAÇÃO FINAL (1).doc: 1373696 bytes, checksum: 6e6f4afc01a6708ec44c276e63213713 (MD5) / Rejected by Ana Hilda Fonseca(anahilda@ufba.br), reason: transformar em pdf. on 2013-06-06T12:59:17Z (GMT) / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-06-06T13:43:50Z No. of bitstreams: 1 Dissertação NORMANDO DISSERTAÇÃO FINAL (1).pdf: 1416415 bytes, checksum: 4927e2bf2b9b19287f6adb42fc059ccf (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-06-06T13:47:50Z (GMT) No. of bitstreams: 1 Dissertação NORMANDO DISSERTAÇÃO FINAL (1).pdf: 1416415 bytes, checksum: 4927e2bf2b9b19287f6adb42fc059ccf (MD5) / Made available in DSpace on 2013-06-06T13:47:50Z (GMT). No. of bitstreams: 1 Dissertação NORMANDO DISSERTAÇÃO FINAL (1).pdf: 1416415 bytes, checksum: 4927e2bf2b9b19287f6adb42fc059ccf (MD5) Previous issue date: 2012 / CAPES / O objetivo deste trabalho foi o desenvolvimento de um procedimento analítico empregado Cromatografia Líquida Ultra Rápida acoplada a Detector de Fluorescência (UFLC-FLU) para determinação de interferentes endócrinos (IEs; bisfenol A (BPA), 4n-nonilfenol (4NP), 4-octifenol (4OP), 4-t-octifenol (4TOP), estriol (E3), estrona (E1), 17β- estradiol (E2) e 17α-etinilestradiol (EE2)) em água do mar. Foi utilizado um sistema de pré-concentração em fase solida (cartucho SPE com fase estácionária C18) para extração e pré-concentração dos IE em as amostras de água do mar. A separação foi otimizada e realizada em um tempo total de corrida de 10 min, em uma coluna cromatográfica Shim-pack XR-ODS C-18 (2,0 mm ID x 50 mm), com a fase móvel de acetonitrila e água ultra pura com gradiente de eluição. A vazão foi de 0,12 mL min-1, a temperatura da coluna foi mantida em 60°C e os comprimentos de onda de emissão e excitação foram de 306 nm e 280 nm, respectivamente. O método validado foi aplicado em amostras de água coletadas na Baia de Todos os Santos, Bahia, Brasil. As amostras foram coletadas na Baía da Ribeira, Feira do São Joaquim, Santo Amaro da Purificação, São Francisco do Conde, Cachoeira e Acupe. As recuperações para o IE variaram entre 84,9% (para o composto 4nOP) e 104% (para o composto 4nNP), e a repetibilidade foi adequada (RSD < 4,5%). Os limites de detecção e quantificação encontrados para os compostos estudados variaram de 4 a 27 µg L-1 e de 19 a 185 µg L-1, respectivamente para o método cromatográfico. Considerando o fator de concentração de 2000 vezes, o LD e LQ variaram de 2 a 23 ng L-1 e de 9 a 96 ng L-1, respectivamente quando calculados para amostra real. Foi observada a ocorrência dos seguintes IE nas amostras reais: bisfenol A (BPA), 17β-estradiol (E2), estriol (E3) e 4n-octifenol (4NOP) em concentrações que variaram de 5 ng L-1 em Santo Amaro a 18,3 ng L-1 em Cachoeira para o E2, 20 ng L-1 do 4NOP na Ribeira a 135 ng L-1 no estuario do Rio Subaé (Santo Amaro), sendo o 4NOP o contaminante onipresente nas amostras analisadas. A concentração de 38 ng L-1 de E3 foi encontrada apenas nas amostras da Ribeira. A presença de BPA foi detectada em quase todas as amostras (o BPA não foi detectado na Ribeira), em níveis entre 13 ng L-1 no estuário do rio Paraguaçu em Cachoeira e aproximadamente 77 ng L-1 no estuário do rio Subaé. Os resultados sugerem que as regiões estudadas encontram-se possivelmente impactadas em relação os IE estudados e que as concentrações encontradas poderiam indicar possíveis danos ao ecossistema marinho local. O método analítico empregando SPE e UFLC-FLU se mostrou eficiente na determinação dos oito compostos de interesse usando um volume de amostra de 4 litros. / Salvador

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-21T18:28:49Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) FABIO DE SOUZA BARBOSA.pdf: 1689958 bytes, checksum: 1fe429224460761440aa3dfeb3cbe066 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-21T18:29:20Z (GMT) No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) FABIO DE SOUZA BARBOSA.pdf: 1689958 bytes, checksum: 1fe429224460761440aa3dfeb3cbe066 (MD5) / Made available in DSpace on 2016-09-21T18:29:20Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) FABIO DE SOUZA BARBOSA.pdf: 1689958 bytes, checksum: 1fe429224460761440aa3dfeb3cbe066 (MD5) Previous issue date: 2015-07-17 / O doripenem é um antibiótico β-lactâmico de amplo espectro de ação. Pertencente ao grupo das carbapenemas, caracteriza-se por apresentar elevada potência e atividade frente à cepas Gram-negativas, produtoras de β-lactamases de espectro estendido (ESBL) e β-lactamases ampC. Apesar de sua grande importância clínica, os antibióticos carbapenêmicos não apresentam boa estabilidade quando em solução, o que é demonstrado em diversos trabalhos descritos na literatura científica. Para o doripenem, há vários trabalhos descritos na literatura que ressaltam sua importância clínica e sua atividade antibiótica. Porém, nota-se a escassez de trabalhos que enfoquem sua estabilidade físico-química, seus produtos e suas rotas de decomposição. O presente trabalho tem como objetivo a validação de um método analítico indicativo de estabilidade por ultra fast liquid chromatography (UFLC), e a avaliação da estabilidade do doripenem em solução, quando submetido a estresse térmico, oxidativo, fotólise e hidrólise em meio ácido e meio alcalino, e determinação da cinética química de decompsição. Para proposição da estrutura química dos produtos de degradação, foram realizadas análises por cromatografia líquida com detecção por espectrometria de massas (LC-MS). O método cromatográfico descrito neste trabalho demonstrou-se adequado para determinação do doripenem na forma de pó para solução injetável, possuindo performance indicativa de estabilidade. A faixa linear do método foi de 5,0 a 40,0 μg/mL, sem desvios de linearidade, sendo estatisticamente comprovada por meio de ANOVA. Com o auxilio do desenho experimental de Plackett–Burman, o método demonstrou-se robusto frente a uma série de fatores. O estudo de degradação forçada demostrou a susceptibilidade do doripenem a diversos fatores de degradação, com acentuada instabilidade frente à hidrólise ácida e alcalina, observando-se degradação aproximada de 60% do seu teor em apenas 2 minutos sob condições alcalinas. A decomposição oxidativa do fármaco seguiu uma cinética de segunda ordem, com constante de velocidade de reação de 0,000086 e 0,00010%-1.min-1, quando submetido à degradação em H2O2 a 3 e 10%, respectivamente. A decomposição térmica apresentou uma energia de ativação de aproximadamente 15 Kcal/mol, valor característico de reações de hidrólise. E na análise cromatográfica com detecção por espectrometria de massas, observou-se que os principais produtos de degradação formados sob condições de termólise e oxidação, apresentam massas moleculares semelhantes, sendo possível a proposição da estrutura química dos mesmos. / Doripenem is a β-lactam antibiotic with a broad spectrum of antimicrobial activity, including gram-negative strains, and producers of extended spectrum β-lactamases (ESBL) and ampC β-lactamases ampC. Despite its great clinical importance, carbapenems do not show good stability when incorporated as solution, as reported in several studies. In reference to doripenem, several works have describing its clinical use, efficacy data and cases of resistance. However, few works mention the drug stability, in terms of degradation products and routes of decomposition. The present work aimed to develop and validate a stability-indicating method by ultra-fast liquid chomatograph (UFLC) for doripenem in powder for injection, purposing an evaluation of stability of reconstituted solution using stress conditions of heat, oxidation, acid hydrolysis, alkaline hydrolysis and photolysis. The chemical kinetic of decomposition was also assayed for thermal and oxidative degradation. For identification of degradation products, the degraded samples where submitted to analysis by LC-MS. The chromatographic method described here proved to be stability-indicating and suitable for the determination of doripenem in drug formulation. The method linearity was performed in the range of 5 to 40 g mL-1, whose correlation coefficient (r) was 0.9999. The robustness testing, assayed against a variety of factors, allowed verifying that the method accept small variations in routine analysis. The forced degradation demonstrated the susceptibility of doripenem to several decomposition factors, being intense the instability to acidic and alkaline hydrolysis. In basic media, the drug residual content was approximately 40% in 2 minutes. At oxidative decomposition, the drug follows second-order kinetics, with a rate reaction of 0.000086 and 0.00010 %-1 min-1, respectively for H2O2 at 3.0 and 10.0 %. The thermal decomposition showed an activation energy of 15 kcal mol-1, a characteristic value for hydrolysis reactions. The analysis by LC-MS revealed that the major degradation products formed under oxidizing conditions and thermolysis present molecular weight (411, 427, 437, 634, 650 and 664). The stability of doripenem must be carefully observed, mainly after reconstitution and storage in adverse conditions of temperature.

Over the past decade, synthetic piperazines have been associated with multiple fatalities and was one of the top 25 identified drugs in 2011. While circumventing legislative controls and preventing the detection in standard drug tests, synthetic piperazine derivatives are encountered in forensic casework as “legal” alternatives to ecstasy (3,4-methylenedioxymethamphetamine). These chemically-produced compounds share very similar pharmacological and psychological effects with ecstasy which in turn has led to their popularity as “party pills”. The long-lasting duration of synthetic piperazines, especially when 1-benzylpiperazine (BZP) is mixed with 1-(3-trifluoromethylphenyl)-piperazine (TFMPP), has also made them desirable to drug users to receive enhanced hallucinogenic effects. Although most methods are optimized to accurately quantify the amount of drugs in biological specimens submitted for forensic toxicology testing, unforeseeable challenges may arise to complicate the analysis such as postmortem redistribution, enzymatic reactions, the presence of bacterial activities, chemical and matrix interferences as well as the lack of reference materials. Thus, the purpose of this research was to investigate the stability of synthetic piperazines in human whole blood under various storage conditions and time ranges. A total of eight synthetic piperazines were assessed on their degrees of degradation using a Shimadzu Ultra-Fast Liquid Chromatography (UFLC) with SCIEX 4000 Q-Trap Electrospray Ionization Tandem Mass Spectrometry in positive ionization mode. These analytes included: 1-benzylpiperazine (BZP), 1-(4-fluorobenzyl)-piperazine (FBZP), 1-(4-methylbenzyl)-piperazine (MBZP), 1-(4-methoxyphenyl)-piperazine (MeOPP), 1-(para-fluorophenyl)-piperazine (pFPP), 1-(3-chlorophenyl)-piperazine (mCPP), 2,3-dichlorophenylpiperazine (DCPP), and 1-(3-trifluoromethylphenyl)-piperazine (TFMPP). Individual unknown samples were prepared by spiking certified reference standards (Cayman Chemical, Ann Arbor, MI, U.S.A.) of each synthetic piperazine into certified drug-free human whole blood (UTAK Laboratories, Inc., Valencia, CA, U.S.A.) independently at 1000 ng/mL. To closely monitor the stability of each compound and potential drug-drug interactions, mixed samples consisted of all eight piperazines were also stored at room temperature (~20°C), 4°C and -20°C for one, three, six, nine and twelve months in dark sealed containers. Solid phase extraction (SPE) was performed to remove unwanted components prior to the injection into the LC system. Drug of Abuse (DAU) mixed-mode copolymeric columns (Clean Screen®, UCT Inc., Levittown, PA, U.S.A.) were utilized with a positive pressure manifold rack followed by evaporating to dryness with low heat at 65°C. All samples were then reconstituted with 250 µL of 50:50 mixture of methanol and 2mM ammonium formate buffer with 0.2% formic acid (Fisher Scientific, Waltham, MA, U.S.A.). Analysis was performed in triplicate using a reversed-phase column (Kinetex® F5, Phenomenex®, Torrance, CA, U.S.A.) with a binary gradient of a 2mM ammonium formate buffer with 0.2% formic acid and methanol with 0.1% formic acid. The total run time was 11.5 minutes including equilibration and the flow rate was 0.4 mL/min. Three internal standards including BZP-d7, mCPP-d8 and TFMPP-d4 (Cerilliant, Round Rock, TX, U.S.A) were used to generate calibration curves that were ranged from 20 ng/mL to 2000 ng/mL. Results revealed that BZP, MBZP and FBZP were more stable than phenyl piperazines over time under all storage conditions, in which MBZP was consistently more stable and still had more than 70% remaining after 12 months. Data showed a smaller degree of degradation when samples were kept frozen or refrigerated; whereas storing at room temperature should be avoided to ensure minimal degradation and detrimental impacts on stability of piperazine compounds. For crime laboratories that are facing backlog situations, case samples with synthetic piperazines should be kept frozen or refrigerated even for time period as short as 30 days or less. However, storing them for too long will clearly affect the quantitation accuracy because phenyl piperazines are more susceptible to degrade completely after six months regardless of storage conditions. Additionally, matrix interference was present due to the outlier of MBZP quantified on Day 270. Drug-drug interaction was also observed in the analyte mixture but the exact stability pattern of phenyl piperazines when mixed together could not be determined from this data set alone due to discrepancies observed on Day 91 and 270. This research project had shown a solid method to examine how quickly or slowly synthetic piperazines degrade in blood at different storage conditions. To further this study, it would be also important to evaluate the number of freeze-thaw cycles on each specimen in order to minimize the effect of non-metabolic degradation.

Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2016-04-07T13:49:01Z No. of bitstreams: 1 Luciane Bastos tese final.pdf: 3085064 bytes, checksum: 1832de4bdbd8e125b7f4a883a01cb07d (MD5) / Approved for entry into archive by Ana Hilda Fonseca (anahilda@ufba.br) on 2016-05-10T17:26:20Z (GMT) No. of bitstreams: 1 Luciane Bastos tese final.pdf: 3085064 bytes, checksum: 1832de4bdbd8e125b7f4a883a01cb07d (MD5) / Made available in DSpace on 2016-05-10T17:26:20Z (GMT). No. of bitstreams: 1 Luciane Bastos tese final.pdf: 3085064 bytes, checksum: 1832de4bdbd8e125b7f4a883a01cb07d (MD5) / A fritura é um processo rápido de preparação do alimento, resultando em produtos com aroma, sabor e textura agradável. As alterações que se processam em um óleo comestível durante o preparo de alimentos, são derivadas de reações hidrolíticas, térmicas e oxidativas. Entre os compostos formados durante a degradação dos óleos pode-se destacar os hidroxialquenais α, β- insaturados, devido a seus efeitos citotóxicos e mutagênicos, uma vez que eles podem interferir em uma ampla gama de atividades biológicas, incluindo a inibição de proteínas e a síntese de DNA. Esse trabalho teve como objetivo estudar a formação de compostos carbonílicos (CC), com ênfase nos hidroxialdeídos α, β- insaturados, na fração líquida do óleo quando submetido a aquecimento contínuo a temperatura típica de processo de fritura (180°C), na presença dos íons metálicos Fe(III), Cu(II) e Mn(II), em diferentes concentrações (10, 30 e 50 g.kg-1 de óleo) ao longo de oito horas de aquecimento.Para determinação dos CC, foi desenvolvida, validada e aplicada metodologia analítica por extração líquido-líquido e análise por UFLC-DAD-ESI-MS. No desenvolvimento do método foram avaliados: solvente extrator, volume do extrator, modo e velocidade de agitação e tempo de sonicação. Na validação do método foram avaliados o efeito da matriz, sensibilidade, seletividade, linearidade das curvas analíticas, limites de detecção e quantificação, precisão e exatidão. Dentre os CC estudados foram identificados e quantificados dez: 4-hidroxi-2-trans- hexenal, 4-hidroxi-2-trans-nonenal, 4,5-epoxi-2-decenal, acroleína, 2-heptenal, 2,4- heptadienal, 2-octenal, 2,4-decadienal, 2-decenal e 2-undecenal. Além destes, o hidroxi-decadienal e o dihidroxi-decenal foram propostos como também sendo formados, baseando-se na análise dos espectros de massas dos picos cromatográficos. Durante o aquecimento, 4-hidroxi-2-trans-nonenal, 2,4-decadienal e 2,4-heptadienal foram os compostos que apresentaram as maiores concentrações médias nas oito horas de aquecimento. No estudo da influência dos metais, de um modo geral, a adição dos íons metálicos às amostras provocou um aumento nas concentrações dos CC, sendo que o 2-undecenal, 4-hidroxi-2-trans-hexenal e o 2- decenal foram os que tiveram os maiores aumentos relativos nas concentrações. Em termos de efeito catalítico, o cobre foi o metal que mostrou-se mais eficiente em promover um aumento nas concentrações, seguido do manganês, sendo que, em geral, esses efeitos eram superiores na concentração de 50 g.kg-1. Por outro lado, os íons metálicos estudados não afetaram significativamente a ordem de concentração com que os CC foram formados; entretanto, atuaram como catalisadores de reações oxidativas, acelerando o processo de degradação dos óleos vegetais e com isso aumentaram a formação de compostos secundários, como os compostos carbonílicos. Finalmente, cabe destacar as concentrações relativamente altas do 4-hidroxi-2-trans-nonenal, 2,4-decadienal e 4,5-epoxi-2- decenal, encontradas neste estudo na fração líquida do óleo de soja, tendo em vista sua alta toxicidade relatada e uma possível transferência para os alimentos em contato com o óleo. / The frying is a rapid process of preparing the food, resulting in products with nice aroma, flavor and texture. The changes that occur in the edible oil during food preparation are derived from hydrolytic, thermal and oxidative reactions. Among the compounds formed during the degradation of oils can highlight the α, β- unsaturated hydroxyalkenals, due to their cytotoxic and mutagenic effects, since they may interfere with a wide range of biological activities, including inhibition of protein and DNA synthesis. This work aimed to study the formation of carbonyl compounds, with emphasis on α, β- unsaturated hydroxyaldehydes present in soybean oil when subjected to the process continuous heating at typical frying temperature (180°C), in the presence of metal ions such as Fe (II), Cu (II) and Mn (II), in different concentrations (10, 30 and 50 g kg-1 oil) over eight hours of heating. For determination of carbonyl compounds in oil samples was developed, validated and applied the analytical method by liquid-liquid extraction and analysis by UFLC- DAD/ESI-MS. In developing the method were evaluated: solvent extractor, extractor volume, mode and rate of stirring and sonication time; in the validation of the method were evaluated the matrix effect, sensitivity, selectivity, linearity of the calibration curves, limits of detection and quantification, precision and accuracy. We identified and quantified ten CC: 4-hydroxy-2-trans-hexenal, 4-hydroxy-2-trans-nonenal, 4,5- epoxy-2-decadal, acrolein, 2-heptenal, 2,4-heptadienal, 2-octenal, 2,4-decadienal, 2- decadal and 2-undecenal. In addition, the hydroxy-dihydroxy-decadienal and decadal also been proposed as being formed, based on the analysis of the mass spectra of the chromatographic peaks. During heating, 4-hydroxy-2-trans-nonenal, 2,4- heptadienal and 2,4-decadienal were compounds which had the highest average concentration within eight hours of heating. In the study of the influence of metals, in general, the addition of metal ions to the samples caused an increase in the concentration of CC, wherein the 2-undecenal, 4-hydroxy-2-hexenal and trans-2- decadal were that had the greatest relative increases in concentrations. In terms of catalytic effect, copper is the metal which was more efficient at promoting an increase in concentration followed by manganese, and, in general, these effects were higher in concentration of 50 g kg-1. On the other hand, the metal ions studied did not significantly affect the order that the CC concentration were formed; however, acted as catalysts for oxidative reactions, accelerating the process of degradation of vegetable oils and thereby increased the formation of secondary compounds, such as carbonyl compounds. Finally, we highlight the 4,5-epoxy-2-decadal relatively high concentrations of HNE, 2,4-decadienal, and in this study the liquid fraction of soybean oil, given its high toxicity and reported a possible transfer to food in contact with the oil.

"Benzo Fury" compounds and derivatives are enactogens similar to 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in various aspects. These compounds are similar in structure to MDMA and MDA, as well as, elicit similar effects such as elevated mood, euphoria and hallucinations. This similarity in effect increases the potential for abuse as MDMA has become less prevalent in some regions as the use of these new psychoactive substances (NPSs) has increased. The benzofury compounds are used as legal alternatives to MDMA because of their marketing as “not for human consumption”. With the relative ease in obtaining NPSs via the Internet, it is possible that these drugs may soon be prevalent in the United States. The project’s goal was to separate, detect, and quantitate the benzofury compounds and derivatives as well as MDA and MDMA in one method of analysis using ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS). The project also examined which method of sample preparation is more effective for these compounds. Six benzofury compounds were researched: 5-(2-aminopropyl)benzofuran) (5-APB), 6-(2-aminopropyl)benzofuran) (6-APB), 5-(2-aminopropyl)-2,3-dihydrobenzofuran (5-APDB), 6-(2-aminopropyl)-2,3-dihydrobenzofuran (6-APDB), 1-(benzofuran-5-yl)-N-methylpropan-2-amine (5-MAPB) and 1-(benzofuran-6-yl)-N-methylpropan-2-amine (6-MAPB) as well as MDMA and MDA. These drugs were analyzed in blood. A liquid-liquid extraction (LLE) method and solid phase extraction (SPE) method were examined to determine which would be better for the separation, detection and quantitation of the benzofury compounds. For the development of the overall method, accuracy, precision, calibration curve, carryover, limit of detection, limit of quantitation, analyte stability, and recovery were examined. The accuracy of the methods examined was greater than +/- 20%. For most analytes, the precision within-run and between-run did not exceed 20%, regardless of the sample preparation method used. A weighting of 1/x was applied to the calibration curve regardless of sample preparation method utilized. The carryover was less than 2% with the SPE method having less carryover (0.02% to 0.50%) than the LLE method (0.05% to 1.56%). The limit of quantitation was determined to be greater than 10 ng/mL. While this was unexpected, the limit of detection calculations determined that this was correct. Using the LLE method in combination with the UFLC-MS/MS method developed, the limit of detection was determined to be at least 9.98 ng/mL. Compared to the LLE method, the SPE limit of detection was lower and calculated to be 3.75 ng/mL. The percent recovery was examined for each of the analytes. It was determined that the SPE was capable of recovering 80% or more of the benzofury compounds and derivatives regardless of the concentration level. The LLE was not as successful in the recovering the benzofury compounds, the best recovery occurred at the 200 ng/mL level with only 65% or less recovered. Analyte stability exhibited a general decrease with variation prior to day 7 and then remains relatively stable until day 14. It was anticipated that the quantitation of the drugs might be complicated due to the similarity in structure between the isomers as well as the similarity of structure between all of the compounds. While this may still be the case, the difficult separation resulted in a re-evaluation and alterations to the UFLC-MS/MS method to correct for these issues. With the change in the UFLC-MS/MS method, further method optimization is required to achieve the appropriate accuracy and limit of quantitation. It was found that the best combination of sample preparation and detection of the benzofury compounds and derivatives is to use SPE followed by an UFLC-MS/MS method.

Made available in DSpace on 2014-12-17T14:16:37Z (GMT). No. of bitstreams: 1 WanessaAB_DISSERT.pdf: 3799185 bytes, checksum: ce77744774059676a04175b60c73d121 (MD5) Previous issue date: 2014-03-26 / The synthetic guanylhydrazones WE010 (3,5-di-tert-butil-4-hidroxibenzaldehyde-guanylhydrazone), WE014 (4-bifenilcarboxialdehydeguanylhydrazone) and WE017 (3,4-diclorobenzaldehydeguanylhydrazone) showed high cytotoxic activity in terms of percentage inhibition of cancer cells growth. However, further progress in the development of these drug candidates requires precise and convenient methods for their qualitative and quantitative analyses. The aim of this study was to develop and validate High Performance Liquid Chromatography with diode-array detection (HPLC-DAD) and Ultra Fast Liquid Chromatography with diode-array detection (UFLC-DAD) methods suitable for as simultaneous as isolated determination of studied guanylhydrazones, based on the optimization of chromatographic parameters and obtaining reduced detection times. The chromatographic analyses of analytes by HPLC were performed on C18 ACE analytical column (150 mm x 4.6 mm), with a particle size of 5.0 ?m. Among all the conditions assayed, the best results of separation were obtained with a mixture of methanol:water (60:40, v/v) as the mobile phase at a flow rate 1.5mL/min and pH of 3.5 adjusted at acetic acid. The UFLC method was developed by experimetal desing techniques in order to find optimal chromatographic analytical conditions, which were achieved on XR-ODS analytical column (50 mm x 3.0 mm), with a particle size of 2,2 ?m, maintained at 25 ?C. The mobile phase was consisted of methanol:water (65:35, v/v) with 0.1% triethylamine (TEA) and pH of 3.5 adjusted at acetic acid, at a flow rate 0.5 mL/min. The procedure were validated following evaluating parameters such as specificity, linearity, limits of detection (LD) and quantification (LQ), precision, accuracy and robustness, giving results within the acceptable range. Although the UFLC method shows better sensitivity (lower values of LD and LQ), robustness (lower rates of relative standard deviation) and minimize spending time and solvent, both developed methods were adequately applied to the analysis of guanylhydrazones molecules, may be used in routine of quality control laboratories. Keywords: guanylhydrazones, HPLC/DAD, UFLC/DAD, validation of analitical method / As guanilhidrazonas sint?ticas WE010 (3,5-di-tert-butil-4-hidroxibenzalde?do-guanilhidrazona), WE014 (4-bifenilcarboxialde?doguanilhidrazona) e WE017 (3,4-diclorobenzalde?doguanilhidrazona) apresentaram alta atividade citot?xica em rela??o a inibi??o do crescimento de c?lulas cancer?genas. Contudo, o avan?o no desenvolvimento desses candidatos a f?rmacos necessitam de m?todos precisos para suas adequadas an?lises quantitativas e qualitativas. O objetivo desse estudo foi desenvolver e validar m?todos por Cromatografia L?quida de Alta Efici?cia com Detector de Arranjo Diodo (CLAE-DAD) e Cromatografia L?quida de Ultra Efici?cia com Detector de Arranjo Diodo (CLUE-DAD) adequados para a determina??o simult?nea, bem como isolada das guanilhidrazonas em estudo, baseado na otimiza??o de par?metros cromatogr?ficos e obten??o de tempos reduzidos de detec??o. As an?lises cromatogr?ficas por CLAE foram realizadas numa coluna anal?tica C18 ACE (150 mm x 4,6 mm), com tamanho de part?cula de 5,0 ?m. Dentre as condi??es analisadas, os melhores resultados de separa??o foram obtidos com uma fase m?vel composta por metanol:?gua (60:40), em um fluxo de 1,5 mL/min. e um pH de 3,5 ajustado com ?cido ac?tico. O m?todo por CLUE foi desenvolvido a partir de t?cnicas de planejamento fatorial, com o objetivo de se encontrar as melhores condi??es anal?ticas, que foram obtidas a partir de an?lise em uma coluna XR-ODS (50 mm x 3,0 mm), com tamanho de part?cula de 2,2 ?m, mantida a 25 ?C. A fase m?vel foi constitu?da por metanol:?gua (65:35) com 0,1% de Trietilamina (TEA) e pH de 3,5 ajustado com ?cido ac?tico. Os procedimentos foram validados a partir da avalia??o de par?metros de especificidade, linearidade, limites de detec??o (LD) e quantifica??o (LQ), precis?o exatid?o e robustez, obtendo-se resultados dentro do intervalo aceit?vel. Embora o m?todo por CLUE tenha mostrado melhor sensibilidade (menores valores de LD e LQ), robustez (menores ?ndices de desvio padr?o relativo) e minimizado gastos de tempo e solvente, ambos os m?todos desenvolvidos foram adequadamente aptos para as an?lises das mol?culas de guanilhidrazonas, podendo ser utilizados na rotina de laborat?rios de controle de qualidade