Class of 2010 Abstract / OBJECTIVES: This is a selective systematic review of the methods used to monitor for the ocular adverse effects of chronic hydroxychloroquine use. This was done to describe the screening methods for detecting retinal toxicity and the recommendations for early detection of hydroxychloroquine associated retinopathy. METHODS: A literature search of OVID-MEDLINE and the Evidence Based Medicine (EBM) search database in the AHSL website were performed for the date range October 1999 to October 2009. Articles were selected based on content related to our purpose statement. Each article must have had at least one or more ophthalmic screening test including: fundus photography, Amsler grid, perimetry, color vision, and multifocal electroretinography. RESULTS: The search results returned a total of 67 articles. A total of twelve articles were selected for review. A total of 959 human subjects were studied and 22 patients had reported retinal changes attributed to hydroxychloroquine. Nine of the eleven studies that included mfERG recommended this specific test for monitoring for hydroxychloroquine induced retinopathy. Only six of the studies recommended the frequency of testing. The most common recommendation was to test at baseline and then at least annually. CONCLUSIONS: The results suggest the mfERG is the most sensitive and objective exam for early documentation of toxic retinopathy. In addition baseline and annual testing was suggested most often. The data also suggest that high risk patients be followed more closely as the most severe and irreversible damage occurs in this population.

Thèse d'exercice : Pharmacie : Nantes : 2007. / Bibliogr.

Thesis (M.Sc.)--University of Alberta, 1997. / Submitted to the Faculty of Graduate Studies and Research in partial fulfilment of the requirements for the degree of Master of Science, Department of Medicine. Also available online.

Submitted by Irene Nascimento (irene.kessia@ufpe.br) on 2017-04-10T19:15:40Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese Fabiana Licia Araujo dos Santos.pdf: 3305306 bytes, checksum: e915bed6750342cef5b377cd2e10834e (MD5) / Made available in DSpace on 2017-04-10T19:15:41Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese Fabiana Licia Araujo dos Santos.pdf: 3305306 bytes, checksum: e915bed6750342cef5b377cd2e10834e (MD5) Previous issue date: 2015-05-25 / CNPQ / Associação em Dose Fixa (ADF) é uma combinação de dois ou mais princípios ativos em uma razão fixa de doses em uma mesma forma farmacêutica. Comodidade para o paciente e consequente incremento na adesão ao tratamento são razões aventadas para estimular desenvolvimento de ADF, sendo estas particularmente importantes no tratamento de doenças crônicas. Assim, por associar atividade antiinflamatória, imunomoduladora e antiartritica, Baccharis trimera (Less.) DC. é uma planta promissora para compor uma ADF com o sulfato de hidroxicloroquina (HCQ), como alternativa inovadora para o tratamento da artrite reumatoide. O sulfato de HCQ integra a relação de fármacos recomendados no consenso brasileiro de doenças reumáticas, podendo inclusive ser utilizado durante o período de amamentação. Sendo assim, o objetivo desse estudo é a obtenção de uma forma farmacêutica ADF (cápsulas), utilizando como agentes terapêuticos o extrato seco liofolizado de B. trimera e o fármaco sulfato de HCQ. O extrato foi obtido por infusão e seco por processo de liofilização, apresentando rendimento de 9% (p/p). A técnica de cromatografia líquida de alta eficiência (CLAE) foi utilizada no ensaio de doseamento da droga vegetal e do extrato seco. As amostras apresentaram valores de 9 e 0,2 mg.g-1 de ácidos cafeicos, respectivamente. A técnica de termogravimetria (TG) revelou a eficiência do processo de liofilização, uma vez que não foi observado na curva TG do extrato seco o evento de perda de água na faixa de 27-100ºC, presente no perfil da droga vegetal. Em relação às propriedades de fluxo e tamanho de partícula, o extrato liofilizado apresentou-se como um pó fino, de elevada área superficial e presença de mesoporos, com baixa capacidade de fluxo, demonstrando a necessidade da adição de excipientes que possibilitem a utilização do insumo para a obtenção da forma farmacêutica. Na sequência, o sulfato de HCQ foi caracterizado por diversas técnicas, com destaque para a polarimetria, Microscopia Eletrônica de Varredura (MEV) e Difração de raios-X (DRX), comprovando a proporção equimolar dos isômeros da HCQ na mistura racêmica e a existência de cristais de tamanhos irregulares com formatos predominantemente hexagonais. A partir de um estudo de planejamento fatorial, otimizou-se o método analítico para doseamento do sulfato de HCQ por CLAE, com base na metodologia descrita na Farmacopeia Americana. O novo método provou ser mais rápido e mais eficiente em relação ao método de referência, por promover redução no tempo de retenção e redução no fluxo da fase móvel. Além disso, o método permitiu a quantificação do sulfato de HCQ a partir de um pico de alta resolução, apresentando valor de 1,087 para o fator de cauda, assegurando uma quantificação precisa da matéria-prima enquanto mistura racêmica. Através da obtenção de cinéticas de degradação dinâmica e isotérmica, o sulfato de HCQ revelou um prazo de validade estimado de 622 dias. A compatibilidade do fármaco-excipiente foi avaliada através de calorimetria exploratória diferencial (DSC), seguida de análises complementares, evidenciando incompatibilidade do fármaco com o glicolato de amido sódico e com a lactose. Diante de todos os dados obtidos durante os estudos de pré-formulação, foi possível desenvolver uma forma farmacêutica ADF que apresentou boas propriedades de fluxo e baixo teor de umidade, situação ideal para manter a estabilidade de formulações que contém extrato de origem vegetal. Esse produto inovador pode surgir como um alívio ao desconforto do paciente que muitas vezes utiliza diferentes classes de medicamentos para o controle de doenças crônicas, como a artrite reumatoide. / Fixed-dose combination (FDC) is an association of two or more active ingredients in a fixed ratio of doses in the same pharmaceutical form. Convenience for the patient and consequent increase in treatment adherence are reasons to stimulate development of FDC, which are particularly important in the treatment of chronic diseases. Thus, by associating anti-inflammatory, immunomodulatory and anti-arthritic activity, Baccharis trimera (Less.) DC is a promising plant to compose a FDC with hydroxychloroquine (HCQ ) sulfate, as an innovative alternative for the treatment of rheumatoid arthritis. The HCQ sulfate integrates the list of drugs recommended in the Brazilian consensus of rheumatic diseases, and it can even be used during breastfeeding. Therefore, the aim of this study is to obtain a pharmaceutical form (capsules) of a FDC, using as therapeutic agents the dry extract of B. trimera and the drug HCQ sulfate. The extract was obtained by infusion and dried by lyophilization process, with yield of 9% (w / w). The high-performance liquid chromatography (HPLC) was used in the assay of the herbal drug and its dry extract. The samples exhibited values of 9 and 0.2 mg.g-1 of cafeic acids, respectively. The thermogravimetry (TG) showed the efficiency of the lyophilization process, since it was not observed in the TG curve of the dry extract a water loss event from 27 to 100°C that is present in the herbal drug profile. Whereas flow properties and particle size, the lyophilized extract was characterized as a fine powder with a high surface area and the presence of mesopores, with low flowability, demonstrating the need for the addition of excipients that enable its use as an active pharmaceutical ingredient to obtaining the pharmaceutical form. Further, the HCQ sulfate was characterized by several techniques, especially polarimetry, Scanning Electron Microscopy (SEM) and X-ray Diffraction (XRD), proving the equimolar ratio of the isomers of HCQ in the racemic mixture and the existence of irregular crystal sizes predominantly in hexagonal shapes. The analytical method for determination of HCQ sulfate by HPLC was also optimized using factorial design. The new method has proved to be faster and more efficient compared to the reference method described at United States Pharmacopeia, by promoting a reduction in the retention time and reducing the flow of mobile phase. Furthermore, the method allows the quantification of HCQ sulfate from a peak of high resolution, presenting value of 1.087 for the tailing factor, assuring a precise quantification of the raw material as the racemic mixture. By obtaining dynamic and isothermal kinetics of degradation, HCQ sulfate revealed an estimated shelf life of 622 days. The drug-excipient compatibility was evaluated by differential scanning calorimetry (DSC), followed by complementary tests, highlighting drug incompatibility with sodium starch glycolate and lactose. Considering all the data obtained during the pre-formulation studies, it was possible to develop a FDC pharmaceutical dosage form which showed good flow properties and low moisture content, ideal situation to maintain the stability of formulations containing herbal extract. This innovative product may come as a relief to the discomfort of the patient who often uses different classes of drugs for the control of chronic diseases such as rheumatoid arthritis.

Submitted by TULIO MORANDIN FERRISSE null (tuliomferrisse@gmail.com) on 2016-05-30T18:50:25Z No. of bitstreams: 1 DISSERTAÇÃO - FINAL.doc: 8888832 bytes, checksum: ca99c5983969f9477ae1a74714d6d280 (MD5) / Rejected by Ana Paula Grisoto (grisotoana@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: A versão final da dissertação/tese deve ser submetida no formato PDF (Portable Document Format). O arquivo PDF não deve estar protegido e a dissertação/tese deve estar em um único arquivo, inclusive os apêndices e anexos, se houver. Por favor, corrija o formato do arquivo e realize uma nova submissão. Agradecemos a compreensão. on 2016-05-31T17:11:40Z (GMT) / Submitted by TULIO MORANDIN FERRISSE null (tuliomferrisse@gmail.com) on 2016-06-02T18:27:31Z No. of bitstreams: 2 DISSERTAÇÃO - FINAL.doc: 8888832 bytes, checksum: ca99c5983969f9477ae1a74714d6d280 (MD5) DISSERTAÇÃO_Mestrado_24_05_2016.pdf: 2063661 bytes, checksum: 945e0429ad165f9d8b64a9f65b91534a (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-06-02T19:56:03Z (GMT) No. of bitstreams: 1 ferrisse_tm_me_arafo.pdf: 2063661 bytes, checksum: 945e0429ad165f9d8b64a9f65b91534a (MD5) / Made available in DSpace on 2016-06-02T19:56:03Z (GMT). No. of bitstreams: 1 ferrisse_tm_me_arafo.pdf: 2063661 bytes, checksum: 945e0429ad165f9d8b64a9f65b91534a (MD5) Previous issue date: 2016-03-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O conceito de reação liquenóide ou interface liquenóide foi introduzido na dermatologia para definir diversas doenças inflamatórias da pele que apresentam características histopatológicas similares. Assim como a pele, a mucosa oral é afetada por uma variedade de lesões liquenóides orais (LLOs). As LLOs foram classificadas em: lesão liquenóide de contato; lesão liquenóide a medicamento; doença do enxerto-versus-hospedeiro e lesões liquenóides não classificáveis de aspecto liquen plano-like, como a estomatite ulcerativa crônica (EUC) recentemente descrita. Além da sobreposição de características entres as lesões que compõem este grupo, o líquen plano oral (LPO) representa o principal diagnóstico diferencial. Tradicionalmente, o diagnóstico das LLOs e do LPO depende da associação clínica e histopatológica, mas em vários casos, esta abordagem não oferece um diagnóstico confiável. As realizações de estudos clínicos e laboratoriais podem auxiliar no entendimento da etiopatogenia destas doenças e refinar a capacidade de diferenciar as LLOs. Diante disto, os objetivos específicos deste estudo foram: (1) Realizar uma revisão sistemática sobre EUC com ênfase específica nas características clínicas, histopatológicas e imunopatológicas desta condição recentemente descrita; e (2) Realizar um estudo prospectivo para avaliar as características clínicas e histopatológicas das LLOs. / The concept of lichenoid reaction or lichenoid interface was introduced in dermatology to define various inflammatory diseases of the skin that have similar histopathological features. Just as the skin and oral mucosa is affected by a variety of oral lichenoid lesions (OLLs). OLLs were classified as contact Lichenoides injury; drug Lichenoides injury; chronic graft versus host and not classifiable lichenoid lesions aspect lichen planus-like, such as chronic ulcerative stomatitis (CUS) recently described. Besides the overlapping features among injuries that make up this group, oral lichen planus (OLP) is the main differential diagnosis. Traditionally, the diagnosis of OLLs and the OLP depends on the clinical and histopathological association, but in many cases, this approach does not provide a confident diagnosis. The achievement of clinical and laboratory studies may help to understand the pathogenesis of these diseases and refine the ability to differentiate OLLs. Thus, the specific objectives of this study were: (1) conduct a systematic review of CUS with particular emphasis on clinical, histopathological and immunopathological of this newly described condition; and (2) Conducting a prospective study to evaluate the clinical and histopathological characteristics of OLLs.

To calculate the sensitivity and specificity of multifocal electroretinography (mfERG) in detection of chloroquine and hydroxychloroquine retinal toxicity, 120 eyes of 63 patients were evaluated using the currently recommended diagnostic tests. The results were compared to those of 54 eyes of 28 control subjects. The sensitivity and specificity of mfERG relative to the combination of automated visual fields and optical coherence tomography (the reference test) were calculated to be 87% and 86.5% respectively. However, analysis of the “false positive” cases proved that mfERG was more sensitive than the reference test and the actual sensitivity and specificity values were higher than the results of this study. Reduction of mfERG amplitude was a strong and reliable sign of early retinal toxicity and was correlated with the cumulative dose of hydroxychloroquine. This correlation was not observed with the reference test quantitative values.

Orientador: Andreia Bufalino / Resumo: O conceito de reação liquenóide ou interface liquenóide foi introduzido na dermatologia para definir diversas doenças inflamatórias da pele que apresentam características histopatológicas similares. Assim como a pele, a mucosa oral é afetada por uma variedade de lesões liquenóides orais (LLOs). As LLOs foram classificadas em: lesão liquenóide de contato; lesão liquenóide a medicamento; doença do enxerto-versus-hospedeiro e lesões liquenóides não classificáveis de aspecto liquen plano-like, como a estomatite ulcerativa crônica (EUC) recentemente descrita. Além da sobreposição de características entres as lesões que compõem este grupo, o líquen plano oral (LPO) representa o principal diagnóstico diferencial. Tradicionalmente, o diagnóstico das LLOs e do LPO depende da associação clínica e histopatológica, mas em vários casos, esta abordagem não oferece um diagnóstico confiável. As realizações de estudos clínicos e laboratoriais podem auxiliar no entendimento da etiopatogenia destas doenças e refinar a capacidade de diferenciar as LLOs. Diante disto, os objetivos específicos deste estudo foram: (1) Realizar uma revisão sistemática sobre EUC com ênfase específica nas características clínicas, histopatológicas e imunopatológicas desta condição recentemente descrita; e (2) Realizar um estudo prospectivo para avaliar as características clínicas e histopatológicas das LLOs. / Abstract: The concept of lichenoid reaction or lichenoid interface was introduced indermatology to define various inflammatory diseases of the skin that have similarhistopathological features. Just as the skin and oral mucosa is affected by a variety oforal lichenoid lesions (OLLs). OLLs were classified as contact Lichenoides injury;drug Lichenoides injury; chronic graft versus host and not classifiable lichenoidlesions aspect lichen planus-like, such as chronic ulcerative stomatitis (CUS) recentlydescribed. Besides the overlapping features among injuries that make up this group,oral lichen planus (OLP) is the main differential diagnosis. Traditionally, the diagnosisof OLLs and the OLP depends on the clinical and histopathological association, butin many cases, this approach does not provide a confident diagnosis. Theachievement of clinical and laboratory studies may help to understand thepathogenesis of these diseases and refine the ability to differentiate OLLs. Thus, thespecific objectives of this study were: (1) conduct a systematic review of CUS withparticular emphasis on clinical, histopathological and immunopathological of thisnewly described condition; and (2) Conducting a prospective study to evaluate theclinical and histopathological characteristics of OLLs. / Mestre

Hydroxychloroquine, initially used as an antimalarial, is used as an immunomodulatory and anti-inflammatory agent for the management of autoimmune and rheumatic diseases such as systemic lupus erythematosus. Lately, there has been interest in its potential efficacy against severe acute respiratory syndrome coronavirus 2, with several speculated mechanisms. The purpose of this review is to elaborate on the mechanisms surrounding hydroxychloroquine. The review is an in-depth analysis of the antimalarial, immunomodulatory, and antiviral mechanisms of hydroxychloroquine, with detailed and novel pictorial explanations. The mechanisms of hydroxychloroquine are related to potential cardiotoxic manifestations and demonstrate potential adverse effects when used for coronavirus disease 2019 (COVID-19). Finally, current literature associated with hydroxychloroquine and COVID-19 has been analyzed to interrelate the mechanisms, adverse effects, and use of hydroxychloroquine in the current pandemic. Currently, there is insufficient evidence about the efficacy and safety of hydroxychloroquine in COVID-19.KEY MESSAGES HCQ, initially an antimalarial agent, is used as an immunomodulatory agent for managing several autoimmune diseases, for which its efficacy is linked to inhibiting lysosomal antigen processing, MHC-II antigen presentation, and TLR functions. HCQ is generally well-tolerated although severe life-threatening adverse effects including cardiomyopathy and conduction defects have been reported. HCQ use in COVID-19 should be discouraged outside clinical trials under strict medical supervision.

Le neuroblastome (NB) est une tumeur fréquente et agressive du jeune enfant. Les tumeurs de haut grade de forme métastatique, généralement développées chez l'enfant de plus de 1 an, sont associées à une importante mortalité malgré un traitement lourd incluant une chimiothérapie à haute dose. La chimiorésistance est donc un problème majeur qui caractérise ces NB de haut grade. Une des hypothèses pour expliquer cette chimiorésistance est celle de l’activation de l’autophagie, un mécanisme auquel recourent les cellules afin de faire face aux situations de stress. D’ailleurs, plusieurs études ont démontré que l'autophagie était activée à la suite des thérapies anticancéreuses. Son inhibition pourrait donc représenter une stratégie thérapeutique très intéressante pour le traitement de cancers. Le but de ce projet de recherche a été de mettre en évidence l'importance de l'autophagie dans les cellules du NB ainsi que l'effet de son inhibition sur la sensibilité des cellules du NB à la chimiothérapie conventionnelle. La présence d'autophagie dans les cellules de NB et sa valeur pronostic ont été évaluées par une étude immunohistochimique et par western blot sur 184 tumeurs patient. Ensuite, dans le but de déterminer l'effet de la chimiothérapie conventionnelle sur le niveau d'autophagie, des études in vitro sur 6 lignées cellulaires ont été effectuées utilisant des tests de mesure d'autophagie (MDC, monodanylcadaverine), de viabilité cellulaire (MTT) et de western blot. Celles ci ont été suivies par des tests d'inhibition de l'autophagie par deux méthodes: l’inactivation du gène ATG5 par un lentivirus contenant un shRNA ciblant ATG5 ou de l'hydroxychloroquine (HCQ), un inhibiteur pharmacologique de l’autophagie. Cette inhibition a été testée seule ou en combinaison avec une chimiothérapie conventionnelle dans le but de déterminer le rôle de l'autophagie dans la sensibilisation des cellules de NB à la chimiothérapie. Ensuite, l’intérêt de l’inhibition de l’autophagie a été évalué sur des modèles murins. Enfin, le niveau d'autophagie a été testé dans des cellules souches de NB. Notre étude a démonté que l'autophagie est présente à un niveau basal dans une majorité des NB mais qu'elle ne représentait pas un facteur pronostic dans ce type de tumeur. Les différentes chimiothérapies testées induisent une augmentation de l'autophagie dans les cellules du NB. Les deux tests d'inhibition ont démontré in vitro que l'autophagie participe à la résistance des cellules aux traitements chimiothérapeutiques classiques du NB. Le blocage de l’autophagie in vivo augmente l’efficacité de la chimiothérapie, cependant certaines données associées au traitement par HCQ devront être complétées. Cette étude démontre que l'inhibition de l'autophagie en combinaison avec la chimiothérapie classique représente une approche thérapeutique prometteuse dans le traitement du NB. / Neuroblastoma (NB) is a common tumor in childhood. Despite major advances in treatments, NB still have a poor prognosis with 40% of mortality. Chemoresistance is a major issue that characterizes aggressive NB. This is a consequence of an autophagic mechanism that tumor cells use to overcome stressful situations encountered during treatments. Autophagy has been the subject of several studies showing its activation in response to anticancer therapies. Its inhibition may therefore represent a very interesting therapeutic strategy for cancer treatment. The purpose of this research was to determine the presence of autophagy in NB cells and the effect of autophagy inhibition in sensitizing NB cells to conventional chemotherapy. The presence of autophagy was verified in 184 NB tumors. To determine the effect of conventional chemotherapy on autophagy, the MTT cell viability test and the autophagy measurement test (MDC, monodansylcadaverine) have been used to study 6 NB cell lines in vitro. An immunohistochemical study also allowed the verification of autophagy activation in tumors grown in mouse models. A lentivirus containing a shRNA against ATG5 was used to generate autophagy deficient cells. Using the MTT and MDC tests, we assessed their sensitivity to chemotherapy. In order to determine the effect of HCQ in sensitizing NB cells to chemotherapy, we used HCQ alone or in combination with conventional chemotherapy. This study demonstrated that autophagy is present at a basal level in NB cells. Unlike for LC3, the results showed that Beclin 1 is a factor of poor prognosis. The MTT and MDC tests have shown that vincristine, doxorubicin, cisplatin, temozolomide, rapamycin, and LY294002 induce autophagy in NB cells. Also, immunohistochemical studies showed that cisplatin treatment increased autophagy in vivo in xenograft model of human NB in mice. ATG5 deficient cells showed greater sensitivity to chemotherapy. Furthermore, the use of these cells in mouse models showed an important role of autophagy in tumor progression as well as an increased sensitivity to vincristine. Finally, combination of HCQ with conventional chemotherapy showed an increased sensitivity of NB cells to chemotherapy compared to cells receiving chemotherapy only. This study demonstrates that inhibition of autophagy in combination with conventional chemotherapy is an attractive therapeutic approach for the treatment of NB.

A hidroxicloroquina (HCQ) é um importante fármaco quiral usado, principalmente, no tratamento de artrite reumatóide, lupus eritematoso sistêmico e malária e cujas propriedades farmacocinéticas e farmacodinâmicas são estereosseletivas. Em relação às propriedades farmacocinéticas, alguns estudos prévios indicam que o metabolismo estereosseletivo parece ser função da espécie estudada, o que implica na necessidade de métodos seletivos para a determinação de seus enantiômeros em materiais biológicos. Assim, propôs-se o desenvolvimento e a validação de métodos para análise dos enantiômeros do fármaco inalterado e de seus principais metabólitos em frações microssomais de homogeneizados de fígado de ratos e camundongos. Para tanto foram empregadas as técnicas de eletroforese capilar (CE) e de cromatografia líquida de alta eficiência (HPLC). Inicialmente foi desenvolvido um método por HPLC, em uma etapa, para a quantificação dos enantiômeros de dois metabólitos da HCQ, desetilcloroquina (DCQ) e desetilhidroxicloroquina (DHCQ) em microssomas de fígado de ratos e camundongos. A separação foi efetuada utilizando-se a coluna Chiralpak AD-RH e hexano:isopropanol (92:8, v/v) acrescido de 0,1% de dietilamina como fase móvel. O procedimento de extração líquido-líquido foi utilizado para a preparação das amostras. A metodologia desenvolvida resultou na completa resolução dos enantiômeros da HCQ, DCQ e DHCQ e pode ser considerada adequada, visto que os parâmetros de validação mostraram valores dentro dos limites exigidos na literatura. O segundo método desenvolvido permitiu a quantificação dos enantiômeros dos três metabólitos da HCQ: DCQ, DHCQ e bisdesetilcloroquina (BDCQ) em microssomas de fígado de camundongos. A separação foi efetuada utilizando-se um tubo capilar de sílica fundida e solução do eletrólito tris(hidroximetil)aminometano 100 mmol/L, ajustada a pH 9,0 com ácido fosfórico e acrescida de 1% (m/v) de β –CD - sulfatada e 30 mmol/L de β –CD - hidroxipropilada. O procedimento de extração líquido-líquido foi eficiente para remover interferentes e os parâmetros de validação mostraram valores dentro dos limites exigidos na literatura. Os métodos desenvolvidos foram aplicados no estudo in vitro do metabolismo da HCQ em frações microssomais de fígado dos animais, verificando-se que o principal metabólito formado é o (-)-(R)-DHCQ, para ambas espécies estudadas. / Hydroxychloroquine (HCQ) is an important chiral drug used specially in the treatment of rheumatoid arthritis, systemic lupus erythematosus and malaria, with stereoselective pharmacokinetic and pharmacodinamic properties. Concerning these properties, some previous studies indicate that the stereoselective metabolism seems to be a function of the studied species, therefore selective methods are required for the determination of its enantiomers in biological matrix. Thus, the present work reports the development and validation of methods for the analysis of the enantiomers of HCQ and its main metabolites in microsomal fraction of rats and mice liver homogenates. Capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC) were used for this purpose. Initially, a one-step HPLC method was developed for the quantification of the enantiomers of two HCQ metabolites, desethylchloroquine (DCQ) and desethylhydroxychloroquine (DHCQ) in microsomal fraction of rats and mice liver homogenates. The separation was performed on a Chiralpak AD-RH column using hexane:isopropanol (92:8, v/v) plus 0.1% diethylamine as the mobile phase. Liquid-liquid extraction procedure was used for sample preparation. The developed methodology resulted in the complete resolution of HCQ, DCQ and DHCQ enantiomers and can be considered suitable because the validation parameters are in accordance with the limits established in the literature. The second developed method allowed the quantification of the enantiomers of the three HCQ metabolites, DCQ, DHCQ and bisdesethylchloroquine (BDCQ) in microsomal fraction of mice liver homogenates. The separation was performed using a fused-silica capillary tube and tris (hydroxymethyl) aminometane 100 mmol/L electrolyte solution, adjusted at pH 9.0 with phosphoric acid, containing 1% (w/v) sulfated- β -CD and 30 mmol/L hydroxypropyl- β -CD. The liquid-liquid extraction procedure was efficient to remove interferents and the validation parameters showed values within accordance to the literature. The developed methods were applied in the in vitro metabolism study of HCQ in microsomal fractions of the liver of the animals and it was verified that the main metabolite formed is (-)-(R)-DHCQ for both animal species studied.