Currently, clinical trials in patients are required by most regulatory authorities for the assessment of bioequivalence of topical products where the drug is not intended for systemic absorption. Hence there is a dire need for suitable methods for the assessment of bioavailability and bioequivalence of such products since clinical safety and efficacy studies are expensive, time-consuming and require very large numbers of patients. Except for topical corticosteroid products where the human skin blanching assay/vasoconstrictor assay has been approved by the US FDA for bioequivalence assessment of those products, no other method has been “officially” approved for use in those investigations. However, a few alternative methods such as tape stripping and microdialysis have been pursued and considered to have the potential for use in ioequivalence/bioavailability studies. The human skin blanching assay was used to assess the bioequivalence of commercially available topical products containing 0.05% clobetasol propionate. Both visual and chromameter data were obtained and a commercially available topical corticosteroid product, Dermovate® cream was used as both the “Test” and the “Reference” product. The results indicated that both visual and chromametric assessments were comparable to each other and that either could be used for the assessment of the bioequivalence of topical products containing clobetasol propionate. The screening procedure was optimized to identify potential “detectors” for inclusion in the bioequivalence studies. This resulted in fewer subjects being required in a bioequivalence pivotal study, still having the necessary power to confirm bioequivalence using the human skin blanching assay. Another objective of this research was to re-visit tape stripping and other possible alternative methods such as dermal microdialysis and to optimize these procedures for bioequivalence assessment of topical formulations where the drug is not intended for systemic absorption. In the past few decades, tape stripping has been used to investigate bioavailability/bioequivalence of various topical formulations. This technique involves the removal of the stratum corneum to assess drug penetration through the skin. A draft FDA guidance for tape stripping was initially published but was subsequently withdrawn due to high variability and poor reproducibility. This research project used an optimized tape stripping procedure to determine bioavailability and establish bioequivalence between three commercially available formulations containing 0.05 % m/m clobetasol propionate. Furthermore, tape stripping was validated by undertaking a study to assess the bioequivalence of a 0.05% topical cream formulation (Dermovate® cream) using the same cream as both the “Test” and “Reference” product, in which bioequivalence was confirmed. The findings highlight the potential of tape stripping as an alternative method for the assessment of bioequivalence of clobetasol propionate formulations and may possibly be extended for use in other topical products. Microdialysis is another useful technique that can assess the penetration of topically applied substances which diffuses through the stratum corneum and into the dermis. Microdialysis has previously been successfully used for in vivo bioavailability and bioequivalence assessments of topical formulations. However, the drugs which were under investigation were all hydrophilic in nature. A major problem with the use of microdialysis for the assessment of lipophilic substances is the binding/adherence of the substance to the membrane and other components of the microdialysis system. As a result, this necessitates the development of a microdialysis system which can be used to assess lipophilic drugs. Intralipid® 20% was investigated and successfully utilized as a perfusate to recover a lipophilic topical corticosteroid, clobetasol propionate, in microdialysis studies. Hence, the bioavailability of clobetasol propionate from an extemporaneous preparation was determined in healthy human volunteers using microdialysis. These findings indicate that in vivo microdialysis can be used to assess lipophilic drug penetration through the skin. A novel approach to investigate drug release from topical formulations containing 0.05% clobetasol propionate using in vitro microdialysis was also undertaken. The in vitro findings were found to be in agreement with the results obtained using tape stripping to assess bioequivalence of the same commercially available products, namely Dermovate® cream, Dovate® Cream and Dermovate® ointment. These results indicate the potential to correlate in vitro with in vivo data for bioequivalence assessment of such topical dosage forms.

A aplicação de toxina botulínica A no lado não paralisado (LNP) é feita para tratar a assimetria resultante da paralisia facial (PF). As unidades de toxina onabotulinica A (Ona) e toxina abobotulinica A (Abo) não são equivalentes. Comparou-se a taxa de conversão de 1:3 em pacientes com PF. Cinquenta e cinco pacientes (idade entre 16 e 67 anos, 43 mulheres), com PF de longa duração foram tratados de forma aleatória com a aplicação de Ona (n = 25) ou Abo (n = 30) no LNP. Efeitos adversos, simetria facial, satisfação subjetiva e Índice de Incapacidade Facial (IIF) foram avaliados após 1 e 6 meses. Os resultados mostraram que a incidência de efeitos adversos foi maior com Abo (93,3% vs. 64,0%, p = 0,007). Avaliação Clínica do LNP diminuiu após 1 mês e aumentou novamente aos 6 meses, sem diferenças entre os grupos. A nota do lado paralisado (LP) foi menor no grupo Ona antes do tratamento, mas semelhante em ambos os grupos depois do tratamento. A nota do LP aumentou depois de 1 mês, e aos 6 meses foi ainda maior que a nota de prétratamento em ambos os grupos. A avaliação subjetiva melhorou em todos os momentos em comparação com a nota do pré-tratamento e diferiu entre os dois grupos apenas em 1 mês, quando o grupo Abo ficou um pouco mais paralisado. Índice de Função Física (IFF) e Índice de Bem-Estar Social (IBES), subescalas do Índice de Incapacidade Facial (IFF), entre os dois grupos não foram diferentes. Concluímos que ambas as toxinas reduziram a assimetria de forma eficiente em pacientes com FP. Os efeitos adversos foram maiores com Abo na equivalência de 1:3 / Botulinum toxin A injection into the nonparalyzed side (NPS) is used to treat asymmetry resulting from facial palsy (FP). OnabotulinumtoxinA (ONA) and abobotulinumtoxinA (ABO) units are not equivalent. We compared the conversion ratio of 1:3 in patients with FP. Fifty-five patients (aged 16-67 years, 43 women) with long-standing FP were randomly treated with either ONA (n = 25) or ABO (n = 30) injections into the NPS. Adverse effects, facial symmetry, subjective satisfaction, and Facial Disability Index (FDI) were assessed after 1 and 6 months. The results showed that the incidence of adverse effects was higher with ABO (93.3% vs. 64.0%, p = 0.007). Clinical scores of the NPS decreased after 1 month and increased again at 6 months, with no betweengroup differences. Scores of the paralyzed side were lower in the ONA group before treatment, but similar in both groups thereafter. The paralyzed side scores increased after 1 month, and at 6 months were still higher than the pretreatment scores in both groups. Subjective assessment improved at all time points compared to pretreatment score and differed between the two groups only at 1 month, when the ABO group was a bit too paralyzed. The Physical Function and Social/Well-Being Function subscales of the FDI did not differ between the two groups. We conclude that both toxins efficiently reduced asymmetry in patients with FP. Adverse effects were higher with ABO at an equivalence ratio of 1:3

Introdução: O Programa Global de Avaliação de Risco Informatizado - PAISM, através de um questionário com 91 perguntas, permite prever o risco de 9 grupos importantes de doenças, que afetam as mulheres. O Programa examinou 15.538 mulheres determinando o risco para: câncer de mama, endométrio, colo de útero, ovário e pulmão; osteoporose; endometriose; DST/AIDS e dislipidemias e foram classificadas em baixo, médio e alto risco para estas doenças. Baseado no risco de avaliação, o programa ofereceu para as mulheres conselho individualizado com relação a hábitos e comportamentos. Objetivos: Avaliar os fatores sócio-demográficos, de acordo com diferentes grupos de risco para cada doença; o estilo e hábitos de vida para os riscos de determinadas doenças e o conhecimento da importância da educação para a saúde, o desempenho e a aceitação do programa. Pacientes e Métodos: Foram estudadas retrospectivamente 320 mulheres selecionadas, sem critérios previamente estabelecidos apenas com a condição de ter participado do Programa Global de Avaliação de Risco Informatizado e não ter câncer. A coleta de dados foi realizada a partir de um outro questionário, constituído de 30 questões que foi aplicado no Ambulatório de Ginecologia do Hospital das Clínicas da FMUSP e em ações existentes nas comunidades carentes da região da Cidade de São Paulo. Esta pesquisa foi delineada inicialmente com base em um modelo conceitual-operativo de natureza quantitativa e, posteriormente, qualitativa. Resultados e conclusões: Existem fatores sócio-demográficos e as variáveis atitudinais, que demonstram o estilo de vida da paciente, também, traçam os grupos de alto risco e colaboram para a observação de determinadas doenças. O nível de conscientização das entrevistadas em relação a algumas variáveis de risco é alto, no entanto, essa conscientização nem sempre se reflete em atitudes e esses fatores acabam influenciando o alto risco nas doenças pesquisadas. As entrevistadas afirmam: o programa é bom, o questionário de avaliação é fácil, a interpretação do risco e as orientações de qualidade de vida são muito boas, demonstrando a importância da educação para a saúde. / Introduction: The Global Program of Evaluation of Informed Risks - GPEIR, through one questionnaire with 91 questions, allows foresee the risk of 9 important groups of disease, which affects women. The Program examined 15.538 women determining the risk to: breast cancer, endometrium, uterus col, ovarian and lung; osteoporosis, endometriosis; DST/AIDS and dislipidemias and were classified below, medium and high risk to those illnesses. Based on the risk of the evaluation, this program offered to women individual advises related to habits and behavior. Objectives: Evaluate the socio-demographic factors, in agreement with different groups of risks to each illness; the stile and habits of life to risks of determined illness and the knowledge of the importance of education to healthy, development and acceptation of the program. Patients and methods: 320 women selected had been studied, without criteria established only with the condition of these women have participated in the Global Program of Evaluation of Informed Risks and don\'t have cancer. The collects of data was realized on base of another questionnaire, with 30 questions that were applied in the \"Ambulatório de Ginecologia do Hospital das Clínicas da FMUSP\" and in actions in devoid communities in São Paulo. This research was delineated initially with base on a conceptual-operative model of nature quantity and lately, quality. Results and conclusion: there are socio-demographic factors and variable attitudinal, which demonstrate the stile of life of patients, also, trace the groups of high risk and collaborate to observe determined illness. The level of awareness of the interviewed in relation to some variable of risk is high, however, this awareness nor always reflects in attitudes and this factors finish influencing a high risk on the illness that have been researched. The interviewed affirm: the program is good, the questionnaire is easy, the interpretation of risk and orientations of quality of life are very good, showing the importance of education to healthy.

A aplicação de toxina botulínica A no lado não paralisado (LNP) é feita para tratar a assimetria resultante da paralisia facial (PF). As unidades de toxina onabotulinica A (Ona) e toxina abobotulinica A (Abo) não são equivalentes. Comparou-se a taxa de conversão de 1:3 em pacientes com PF. Cinquenta e cinco pacientes (idade entre 16 e 67 anos, 43 mulheres), com PF de longa duração foram tratados de forma aleatória com a aplicação de Ona (n = 25) ou Abo (n = 30) no LNP. Efeitos adversos, simetria facial, satisfação subjetiva e Índice de Incapacidade Facial (IIF) foram avaliados após 1 e 6 meses. Os resultados mostraram que a incidência de efeitos adversos foi maior com Abo (93,3% vs. 64,0%, p = 0,007). Avaliação Clínica do LNP diminuiu após 1 mês e aumentou novamente aos 6 meses, sem diferenças entre os grupos. A nota do lado paralisado (LP) foi menor no grupo Ona antes do tratamento, mas semelhante em ambos os grupos depois do tratamento. A nota do LP aumentou depois de 1 mês, e aos 6 meses foi ainda maior que a nota de prétratamento em ambos os grupos. A avaliação subjetiva melhorou em todos os momentos em comparação com a nota do pré-tratamento e diferiu entre os dois grupos apenas em 1 mês, quando o grupo Abo ficou um pouco mais paralisado. Índice de Função Física (IFF) e Índice de Bem-Estar Social (IBES), subescalas do Índice de Incapacidade Facial (IFF), entre os dois grupos não foram diferentes. Concluímos que ambas as toxinas reduziram a assimetria de forma eficiente em pacientes com FP. Os efeitos adversos foram maiores com Abo na equivalência de 1:3 / Botulinum toxin A injection into the nonparalyzed side (NPS) is used to treat asymmetry resulting from facial palsy (FP). OnabotulinumtoxinA (ONA) and abobotulinumtoxinA (ABO) units are not equivalent. We compared the conversion ratio of 1:3 in patients with FP. Fifty-five patients (aged 16-67 years, 43 women) with long-standing FP were randomly treated with either ONA (n = 25) or ABO (n = 30) injections into the NPS. Adverse effects, facial symmetry, subjective satisfaction, and Facial Disability Index (FDI) were assessed after 1 and 6 months. The results showed that the incidence of adverse effects was higher with ABO (93.3% vs. 64.0%, p = 0.007). Clinical scores of the NPS decreased after 1 month and increased again at 6 months, with no betweengroup differences. Scores of the paralyzed side were lower in the ONA group before treatment, but similar in both groups thereafter. The paralyzed side scores increased after 1 month, and at 6 months were still higher than the pretreatment scores in both groups. Subjective assessment improved at all time points compared to pretreatment score and differed between the two groups only at 1 month, when the ABO group was a bit too paralyzed. The Physical Function and Social/Well-Being Function subscales of the FDI did not differ between the two groups. We conclude that both toxins efficiently reduced asymmetry in patients with FP. Adverse effects were higher with ABO at an equivalence ratio of 1:3

Introdução: O Programa Global de Avaliação de Risco Informatizado - PAISM, através de um questionário com 91 perguntas, permite prever o risco de 9 grupos importantes de doenças, que afetam as mulheres. O Programa examinou 15.538 mulheres determinando o risco para: câncer de mama, endométrio, colo de útero, ovário e pulmão; osteoporose; endometriose; DST/AIDS e dislipidemias e foram classificadas em baixo, médio e alto risco para estas doenças. Baseado no risco de avaliação, o programa ofereceu para as mulheres conselho individualizado com relação a hábitos e comportamentos. Objetivos: Avaliar os fatores sócio-demográficos, de acordo com diferentes grupos de risco para cada doença; o estilo e hábitos de vida para os riscos de determinadas doenças e o conhecimento da importância da educação para a saúde, o desempenho e a aceitação do programa. Pacientes e Métodos: Foram estudadas retrospectivamente 320 mulheres selecionadas, sem critérios previamente estabelecidos apenas com a condição de ter participado do Programa Global de Avaliação de Risco Informatizado e não ter câncer. A coleta de dados foi realizada a partir de um outro questionário, constituído de 30 questões que foi aplicado no Ambulatório de Ginecologia do Hospital das Clínicas da FMUSP e em ações existentes nas comunidades carentes da região da Cidade de São Paulo. Esta pesquisa foi delineada inicialmente com base em um modelo conceitual-operativo de natureza quantitativa e, posteriormente, qualitativa. Resultados e conclusões: Existem fatores sócio-demográficos e as variáveis atitudinais, que demonstram o estilo de vida da paciente, também, traçam os grupos de alto risco e colaboram para a observação de determinadas doenças. O nível de conscientização das entrevistadas em relação a algumas variáveis de risco é alto, no entanto, essa conscientização nem sempre se reflete em atitudes e esses fatores acabam influenciando o alto risco nas doenças pesquisadas. As entrevistadas afirmam: o programa é bom, o questionário de avaliação é fácil, a interpretação do risco e as orientações de qualidade de vida são muito boas, demonstrando a importância da educação para a saúde. / Introduction: The Global Program of Evaluation of Informed Risks - GPEIR, through one questionnaire with 91 questions, allows foresee the risk of 9 important groups of disease, which affects women. The Program examined 15.538 women determining the risk to: breast cancer, endometrium, uterus col, ovarian and lung; osteoporosis, endometriosis; DST/AIDS and dislipidemias and were classified below, medium and high risk to those illnesses. Based on the risk of the evaluation, this program offered to women individual advises related to habits and behavior. Objectives: Evaluate the socio-demographic factors, in agreement with different groups of risks to each illness; the stile and habits of life to risks of determined illness and the knowledge of the importance of education to healthy, development and acceptation of the program. Patients and methods: 320 women selected had been studied, without criteria established only with the condition of these women have participated in the Global Program of Evaluation of Informed Risks and don\'t have cancer. The collects of data was realized on base of another questionnaire, with 30 questions that were applied in the \"Ambulatório de Ginecologia do Hospital das Clínicas da FMUSP\" and in actions in devoid communities in São Paulo. This research was delineated initially with base on a conceptual-operative model of nature quantity and lately, quality. Results and conclusion: there are socio-demographic factors and variable attitudinal, which demonstrate the stile of life of patients, also, trace the groups of high risk and collaborate to observe determined illness. The level of awareness of the interviewed in relation to some variable of risk is high, however, this awareness nor always reflects in attitudes and this factors finish influencing a high risk on the illness that have been researched. The interviewed affirm: the program is good, the questionnaire is easy, the interpretation of risk and orientations of quality of life are very good, showing the importance of education to healthy.

The thrust of this thesis is to study oral solid dosage formulation using hot melt coating method and to use pharmacokinetic modeling and simulation (PK M&S) as a tool that can help to predict pharmacokinetics of a drug in human and the probability of passing various bioequivalence criteria of the formulation based on the PK of the drug. Hot-melt coating using a new method, direct blending, was performed to create immediate and sustained release formulations (IR and SR). This new method was introduced to offer another choice to produce IR and SR drug delivery formulations using single and double coating layer of waxes onto sugar beads and/or drug loaded pellets. Twelve waxes were applied to coat sugar cores. The harder the wax the slower the drug was released from single coated beads. The wax coating can be deposited up to 28% of the weight of the core bead with 58% drug loading efficiency in the coating The cores were coated with single or double wax layers containing acetaminophen. Carnauba wax coated beads dissolved in approximately 6 hrs releasing 80% of loaded drug. However, when covered with another layer, the drug loaded beads released drug for over 20 hrs. When drug loaded pellets were used as cores, 33-58% drug loading was achieved. Double coated pellets exhibited a near zero order drug release for up to 16 hrs. Hot melt coating by direct blending using waxes is a simple process compared to conventional hot melt coating using coating pan or fluid bed coating machines. It offers an alternative way of making immediate, sustained drug release (IR, SR) and modified release (IR+SR) oral dosage forms of drugs which are stable at high temperature (100°C). The pellet-containing-drug coated formulations provide options when higher drug loading is warranted. It is required by the US Food and Drug Administration (FDA) that a new modified –release (MR) product or identical generic product be regarded as bioequivalent (BE) to the originators reference drug product. However, there are concerns that current regulatory criteria are not sufficient when evaluating bioequivalence (BE) for many MR products, and additional metrics for BE assessment of the products should be applied to ensure therapeutic equivalence. This study used pharmacokinetic modeling and simulation (M&S) to investigate 1) the probability of BE occurring between the MR test and reference products 2) the rates of false positive and true negative of the BE test; and 3) the estimation of the sample size in pivotal BE studies; all of which when partial area under the curves (pAUCs) were applied as additional BE criteria. Reference data of two MR forms of methylphenydate HCl (MPH) were simulated and obtained from literature (formulation Q and Metadate CD, respectively). Monte Carlo simulations were performed to simulate the test drug concentration profiles and BE assessment was carried out utilizing the mean (method 1) and individual concentration time curves (method 2). For formulation Q, adding pAUC₀₋[subscript Tmax] to current BE criteria reduced the possibility of passing BE from approximately 98% to 85%, with a true negative rate of 5%. The earlier the time points used to determine for pAUC before Tmax, the lower the chance of passing BE for the test product. The possibility of passing BE varied and depended on the coefficient of variations (CV) of T[subscript lag], K[subscript a] and K[subscript e] and that considerable variability in the parameters affected the earlier segments of the drug concentration profile curves more. Similar drug concentration time profiles between the test and reference products is recommended to ensure bioequivalence occurs with a reasonable subject sample size. A similar scenario was seen when Metadate CD was used as the reference product. PK M&S can help provide appropriate additional metrics to assure the BE test is a better tool ensuring therapeutic equivalence for MR products with little negative impact to generic manufacturers. Predictions can also be made about the required sample size and the chances of passing BE with any addition to the conventional three criteria for the test product. PK M&S was also used to predict drug concentrations of levofloxacin in tissue. Levofloxacin has been widely used in clinical practice as an effective broad-spectrum antimicrobial, however tendonitis and tendon rupture have been reported with increasing use of this agent. Here, these incidents will be assessed by investigating pharmacokinetic behavior of the compound to see if they are related to drug's tissue disposition. The PK model for levofloxacin was established. Mean concentration time profiles of single or multiple dosing of 500 mg levofloxacin following oral and IV infusion administration were simulated. Monte Carlo simulation was used to simulate the drug concentration time profiles in plasma (compartment 1) and tissue (compartment 2) after seven dosing regimens while varying the drug's elimination and distribution rates to see the effect of changing those rates have on the drug accumulation in tissue. Monte Carlo Simulation shows that low elimination rates affect the drug concentration in plasma and tissue significantly with the level in plasma rising up to 35 μg/mL at day 7. A normal elimination rate together with escalation of distribution rates from plasma to tissue could increase the tissue concentration after 7 doses to 9.5 µg/mL, a value that is more than twice that of normal. PK M&S can be used as an effective tool to evaluate drug concentration in different compartments (plasma and tissues, for example). The unexpectedly high concentration values in some cases may explain, at least in part, the reason of tendinopathy occurs in the clinical setting. / Graduation date: 2012 / Access restricted to the OSU Community at author's request from June 7, 2012 - June 7, 2014

Submitted by Alexandre Sousa (alexandre.sousa@incqs.fiocruz.br) on 2015-04-10T11:28:22Z No. of bitstreams: 1 Tese_Heliana.pdf: 5204865 bytes, checksum: 96e0c7d76a906d0b86274058a747043a (MD5) / Approved for entry into archive by Alexandre Sousa (alexandre.sousa@incqs.fiocruz.br) on 2015-04-10T11:29:14Z (GMT) No. of bitstreams: 1 Tese_Heliana.pdf: 5204865 bytes, checksum: 96e0c7d76a906d0b86274058a747043a (MD5) / Approved for entry into archive by Alexandre Sousa (alexandre.sousa@incqs.fiocruz.br) on 2015-04-10T11:30:15Z (GMT) No. of bitstreams: 1 Tese_Heliana.pdf: 5204865 bytes, checksum: 96e0c7d76a906d0b86274058a747043a (MD5) / Made available in DSpace on 2015-04-10T11:30:15Z (GMT). No. of bitstreams: 1 Tese_Heliana.pdf: 5204865 bytes, checksum: 96e0c7d76a906d0b86274058a747043a (MD5) Previous issue date: 2014 / Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde / Para que um medicamento possa ser registrado como genérico ou para obter seu registro como similar no Brasil é necessária a execução de uma série de testes bioanalíticos regulamentados por legislações e resoluções do Ministério da Saúde. Estes ensaios são chamados de estudos de bioequivalência/biodisponilidade relativa (BD/BE) e só podem ser realizados por centros de pesquisa devidamente habilitados pela Agência Nacional de Vigilância Sanitária - Ministério da Saúde (ANVISA/MS). Os estudos de BD/BE são realizados em seres humanos e devem assegurar a identidade dos produtos em relação à velocidade da absorção e quantidade do fármaco absorvido. De relevância considerável para a saúde pública e de interesses sócio-econômicos, torna-se importante criar mecanismos e ferramentas que possam dar garantias de que os medicamentos a serem lançados no mercado apresentem as mesmas características e propriedades físico-químicas que o medicamento reconhecido como de referência. Sendo assim, faz-se necessário o desenvolvimento de metodologias seguras e dinâmicas de avaliação da garantia da qualidade e dos processos referentes aos estudos realizados. Neste contexto, este trabalho tem como objetivo apresentar uma nova abordagem da garantia da qualidade, referente à etapa bioanalítica de estudos de BD/BE, com enfoque na Qualidade Total, utilizando o estudo de Levodopa + benserazida como piloto. A proposta é de uma avaliação da qualidade que não se limita às avaliações descritas nas normas e legislações específicas, mas avaliar a qualidade do todo. Nosso trabalho demonstrou um modelo de Sistema de Gestão, que proporcionou o aumento da qualidade dos estudos realizados, diminuindo a visão dicotomista entre Qualidade e Laboratório e, consequentemente, a comercialização dos medicamentos genéricos com qualidade assegurada, após avaliação pelo estudo de BD/BE. / For a drug to be registered as generic or to obtain its registration as similar medication in Brazil, a series of bioanalytical tests regulated by laws and resolutions of the Ministry of Health are required. These tests are called bioequivalence studies / relative bioavailability (BE/BA) and can only be carried out by research centers accredited by the National Health Surveillance Agency - Ministry of Health (ANVISA / MS). Studies BE/BA are performed in humans and should ensure the identity of the products relative to the rate of absorption and amount of drug absorbed. Of considerable relevance to public health and socio-economic interests, it is important to create mechanisms and tools that can provide assurance that the drugs to be launched in the market have the same characteristics and physicochemical properties of the drug known as a reference. Therefore, it is necessary to develop secure, dynamic assessment methodologies and quality assurance processes related to the studies performed. In this context, the present work aims to present a new approach to quality assurance regarding the bioanalytical phase of studies BE/BA, with a focus on Total Quality Control, using the study of levodopa + benserazide as a pilot study. The proposal is of a quality assessment that is not limited to the assessments described in the specific rules and laws, but to evaluate the quality of the whole. Our study demonstrated a model management system, which provided the increase in the quality of studies, reducing the dichotomous view between Quality and Laboratory and hence the marketing of generic drugs with assured quality, after evaluation by the study BE/BA.

The widespread acceptance of topical formulations intended for local and/or regional activity has prompted renewed interest in developing a model to determine the bioavailability of drugs in order to establish bioequivalence as a means of evaluating formulation performance of multisource products and also for use during formulation development. Current in vivo techniques such as blister suction and skin biopsy amongst others used to determine the bioavailability and/or bioequivalence of topical formulations are either too invasive to generate appropriate concentration-time profiles or require large numbers of study subjects thereby making the study expensive and time-consuming. Moreover, there are currently no sampling techniques that can demonstrate dermal bioavailability and/or bioequivalence of topical formulations intended for local and/or regional activity. Dermal microdialysis is a relatively new application of microdialysis that permits continuous monitoring of endogenous and/or exogenous solutes in the interstitial fluid. The technique is involves the implantation of semi-permeable membranes which are perfused with an isotonic medium at extremely slow flow rates and collection of microlitre sample volumes containing diffused drugs. Tape stripping, a relatively older technique, has been extensively used in comparative bioavailability studies of various topical formulations. However, due to shortcomings arising from reproducibility and inter-subject variation amongst others, the published FDA guidance outlining the initial protocol was subsequently withdrawn. The incorporation of transepidermal water loss with tape stripping has garnered renewed interest and has been used for the determination of drug bioavailability from a number of topical formulations. Hence the primary objective of this research is to develop and evaluate microdialysis sampling and tape stripping techniques, including the incorporation of the determination of transepidermal water loss, to assess the dermal bioavailability of ketoprofen from topical gel formulations and to develop models for bioequivalence assessment. A rapid UPLC-MS/MS method with requisite sensitivity for the analysis of samples generated from dermal microdialysis was developed and validated which accommodated the microlitre sample volumes collected. An HPLC-UV method was developed and validated for the analysis of samples generated from the in vitro microdialysis and in vivo tape stripping studies. The work presented herein contributes to a growing body of scientific knowledge seeking to develop a model for the determination of bioequivalence of pharmaceutically equivalent topical formulations intended for local and/or regional activity in human subjects.