Prior experimental research clearly reveals that the performance of reinforced concrete frame structures under earthquake loading is closely related to the behavior of beam-to-column connection regions. In order for a reinforced concrete building to have an adequate response under high lateral deformations, beam-to-column connections should be able to preserve their integrity. However, even today beam-to-column connections are assumed to be rigid or elastic, leading to an incorrect estimation of the structural response under earthquake loading. One of the basic reasons for the assumption of rigid joints is the lack of analytical models that adequately represent the seismic behavior of the connection region. In this thesis, an analytical model that realistically represents the beam-to-column connection response is developed, in the light of prior experimental data. The experimental subassemblies used in the generation of the analytical model are later modeled in OpenSees environment in order to verify the accuracy of the model. Throughout the research, utmost attention is paid for the model to be simple enough to be used practically and also to cover a wide range of beam to column connection properties.

<p>It is relatively straightforward to establish equilibrium inmagnetically confined plasmas, but the plasma is frequentlysucceptible to a variety of instabilities that are driven bythe free energy in the magnetic field or in the pressuregradient. These unstable modes exhibit effects that affect theparticle, momentum and heat confinement properties of theconfiguration. Studies of the dynamics of several of the mostimportant modes are the subject of this thesis. The studies arecarried out on plasmas in the reversed field pinch (RFP)configuration.</p><p>One phenomenon commonly observed in RFPs is mode walllocking. The localized nature of these phase- and wall lockedstructures results in localized power loads on the wall whichare detrimental for confinement. A detailed study of the walllocked mode phenomenon is performed based on magneticmeasurements from three RFP devices. The two possiblemechanisms for wall locking are investigated. Locking as aresult of tearing modes interacting with a static field errorand locking due to the presence of a non-ideal boundary. Thecharacteristics of the wall locked mode are qualitativelysimilar in a device with a conducting shell system (TPE-RX)compared to a device with a resistive shell (Extrap T2). Atheoretical model is used for evaluating the threshold valuesfor wall locking due to eddy currents in the vacuum vessel inthese devices. A good correlation with experiment is observedfor the conducting shell device.</p><p>The possibility of succesfully sustaining discharges in aresistive shell RFP is introduced in the recently rebuiltdevice Extrap T2R. Fast spontaneous mode rotation is observed,resulting in low magnetic fluctuations, low loop voltage andimproved confinement. Wall locking is rarely observed. The lowtearingmode amplitudes allow for the theoretically predictedinternal nonresonant on-axis resistive wall modes to beobserved. These modes have not previously been distinguisheddue to the formation of wall locked modes. The internal andexternal nonresonant resistive wall modes grow on the timescale of the shell penetration time. These growth rates dependon the RFP equilibrium. The internal nonresonant resistive wallmodes dominate in Extrap T2R, especially for shallow reverseddischarges. The external nonresonant modes grow solely in deepreversal discharges.</p><p><b>Keywords</b>Nuclear fusion, reversed field pinch, resistiveinstabilities, wall locked modes, tearing modes, resistiveshell modes, field errors, EXTRAP-T2, EXTRAP-T2R, TPE-RX</p>

La description magnétohydrodynamique est utilisée pour étudier les plasmas de fusion par confinement magnétique dans deux configurations: tokamak et reversed field pinch. Une méthode de Fourier pseudo-spectrale et une technique de pénalisation en volume sont employées pour résoudre les équations. La méthode de pénalisation permet d'introduire des conditions aux limites de Dirichlet et donc de faire varier facilement la géométrie considérée. Les simulations dans des géométries toroïdales de type tokamak montrent l'apparition spontanée de vitesses. Une importante composante toroïdale se développe si le système est peu dissipatif. Il est aussi montré que la brisure de symétrie dans la forme de la section du tore fait apparaitre un moment angulaire toroïdal. Pour le Reversed Field Pinch on montre l'émergence de structures hélicoïdales. La forme de ces structures varie en fonction des coefficients de transport ainsi que du paramètre de pincement du champ magnétique imposé. Pour compléter l'étude on compare les résultats du tore aux calculs dans un cylindre périodique. Les différences dans la dynamique des deux cas sont mises en avant. Finalement les simulations sont confrontées à des expériences et un meilleur accord est observé entre simulation et expérience pour la géométrie toroïdale que pour la géométrie cylindrique.

Analytical chemistry is a field with a vast variety of applications. A robust companion in the field is liquid chromatography, the method used in this thesis, which is an established workhorse and a versatile tool in many different disciplines. It can be used for identification and quantification of interesting compounds generally present in low concentrations, called analytical scale chromatography. It can also be used for isolation and purification of high value compounds, called preparative chromatography. The latter is usually conducted in large scale with high concentrations. With high concentrations it is also possible to determine something called adsorption isotherms. Determination of adsorption isotherms is a useful tool for quite a wide variety of reasons. It can be used for characterisation of chromatographic separation systems, and then gives information on the retention mechanism as well as provides the possibility to study column-column and batch-batch reproducibility. If a protein is immobilised on a solid support, adsorption isotherms can be used for pharmacological characterisation of drug-protein interactions. Moreover, they can be used for the study of unexpected chromatographic phenomena. If the adsorption isotherm is known it is also possible to simulate chromatograms, and subsequently optimise the separation process numerically. The gain of a numerically optimised separation process is higher purity or yield of valuable compounds such as pharmaceuticals or antioxidants, as well as reducing the solvent usage. Taken all together, it saves time, money and the environment. However, the process of the adsorption isotherm determination requires a number of careful experimental considerations and preparations, and these are the main focus of the thesis. Important steps along the way include the choice of separation system and of suitable analytes, preparation of mobile phases and sample solutions, calibration, determination of injection profiles and column void, and of course the adsorption isotherm determination method itself. It is also important to keep track of parameters such as temperature and pH. These issues are discussed in this thesis. At the end, a description of useful methods for processing of the raw adsorption isotherm data is presented, as well as a brief passage on methods for numerical optimisation.

The importance of seismic rehabilitation became evident with 1992 Erzincan Earthquake, after which a large number of reinforced concrete buildings damaged in recent earthquakes required strengthening as well as repair. In the studies related to rehabilitation, it has been realized that inadequate lateral stiffness is one of the major causes of damage in reinforced concrete buildings. Recently, economical, structurally effective and practically applicable seismic retrofitting techniques are being developed in METU Structural Mechanics Laboratory to overcome these kinds of problems. The strengthening technique proposed in this thesis is on the basis of the principle of strengthening the existing hollow brick infill walls by using high strength precast concrete panels such that they act as cast-in-place concrete infills improving the lateral stiffness. Also, the technique would not require evacuation of the building and would be applicable without causing too much disturbance to the occupant. For this purpose, after two preliminary tests to verify the proper functioning of the newly developed test set-up, a total of fourteen one-bay two story reinforced concrete frames with hollow brick infill wall, two being unstrengthened reference frames, were tested under reversed cyclic lateral loading simulating earthquake loading. The specimens were strengthened by using six different types of precast concrete panels. Strength, stiffness, energy dissipation and story drift characteristics of the specimens were examined by evaluating the test results. Test results indicated that the proposed seismic strengthening technique can be very effective in improving the seismic performance of the reinforced concrete framed building structures commonly used in Turkey. In the analytical part of the study, hollow brick infill walls strengthened by using high strength precast concrete panels were modelled once by means of equivalent diagonal struts and once as monolithic walls having an equivalent thickness. The experimental results were compared with the analytical results of the two approaches mentioned. On the basis of the analytical work, practical recommendations were made for the design of such strengthening intervention to be executed in actual practice.

A combinação entre o delapril (DEL), um inibidor da enzima conversora de angiotensina e o manidipino (MAN), um antagonista dos canais de cálcio, produz um efeito anti-hipertensivo sinérgico, podendo ser considerado um ótimo tratamento para pacientes com hipertensão essencial leve e moderada. No presente trabalho foram desenvolvidos e validados métodos cromatográfico e eletroforético para avaliação simultânea de DEL e MAN em produto farmacêutico. As análises por cromatografia líquida em fase reversa (CL-FR) foram executadas utilizando coluna C8 (250 mm x 4,6 mm), mantida a 35 oC. A fase móvel foi constituída por acetonitrila e solução de trietilamina 0,3%, pH 3,0 (55:45; v/v), eluída na vazão de 1,2 mL/min com detecção a 220 nm. Paralelamente, desenvolveu-se método por eletroforese capilar, utilizando modo de separação por cromatografia eletrocinética micelar (MEKC) e ácido salicílico como padrão interno. Foi utilizado capilar de sílica fundida (comprimento efetivo de 72 cm) mantido a 35 °C, com solução eletrolítica composta de tampão borato 50 mM e dodecil sulfato de sódio (SDS) 5 mM, pH 9,0. Voltagem de 25 kV foi aplicada e a injeção foi de 50 mbar durante 5 s, com detecção a 208 nm. A especificidade e a capacidade dos métodos serem indicativos de estabilidade foram demonstradas através de estudos de degradação forçada dos fármacos e pela não interferência dos excipientes nas análises. Além disso, o desenho experimental Plackett-Burman foi utilizado para a avaliação da robustez, observando-se resultados adequados para ambos métodos. Os procedimentos foram validados de acordo com guias aceitos internacionalmente, observando-se resultados em uma faixa aceitável. Os métodos propostos foram aplicados com sucesso na determinação quantitativa simultânea de DEL e MAN em comprimidos, não havendo diferença significativa dos resultados (P>0,05), contribuindo, portanto, para aprimorar o controle da qualidade, assegurando a eficácia terapêutica. / The combination of delapril (DEL), an angiotensin converting enzyme inhibitor and manidipine (MAN), an antagonist of calcium channels, produces a synergic antihypertensive effect and may be regarded as an optimal antihypertensive drug treatment in mild to moderate essential hypertensive patients. The chromatographic and eletrophoretic methods for the simultaneous evaluation of DEL and MAN in pharmaceutical product were developed and validated in the present work. The reversed-phase liquid chromatography (RP-LC) method was carried out on a C8 column (250 mm x 4.6 mm i.d., 5 μm), maintained at 35 ºC. The mobile-phase consisted of acetonitrile and a solution of triethylamine 0.3% pH 3.0 (55:45; v/v), running at a flow rate of 1.2 mL/min, with detection at 220 nm. The capillary electrophoresis method was developed using the micellar electrokinetic chromatography (MEKC) as the separation mode, and salicylic acid as internal standard. The analysis were performed on a fused-silica capillary (effective length of 72 cm) maintained at 35 °C, with 50 mM of borate buffer and 5 mM of sodium dodecyl sulfate (SDS) at pH 9.0 as background electrolyte. The separation was achieved at 25 kV applied voltage and the injection was performed at 50 mbar for 5 s, with detection at 208 nm. The specificity and stability-indicating capability of the methods were demonstrated through forced degradation studies, which also show that there is no interference of the excipients in the analysis. Moreover, the Plackett- Burman experimental design was used for robustness evaluation, giving acceptable results for both methods. The procedures were validated according to Internationals guidelines, giving results within the acceptable range. Therefore, the proposed methods were successfully applied for the simultaneous quantitative analysis of DEL and MAN in the tablet dosage form, showing non-significant difference (P>0.05), contributing to improve the quality control and to assure the therapeutic efficacy.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Interleukin 11 (IL-11) is a multifunctional cytokine in the IL-6 type family of long-chain helical cytokines, which modulates the proliferation, differentiation and maturation of various types of hematopoietic cells. Recombinant human interleukin-11 (rhIL-11) produced by DNA technology in Escherichia coli is currently being used worldwide for the prevention of thrombocytopenia and to reduce the need for platelet transfusions after myelosuppressive chemotherapy in patients with nonmyeloid malignancies. A stability-indicating reversed-phase liquid chromatography (RP-LC) method was validated for the assessment of recombinant human interleukin-11 (rhIL-11) in biopharmaceutical formulations. The RP-LC method was carried out on a Jupiter C4 column (250 mm x 4.6 mm i.d.), maintained at 25ºC. The mobile phase A consisted of 0.1% TFA and the mobile phase B was acetonitrile with 0.1% TFA, run as follows: time 0 to 0.1 min 40% of B; from 0.1 to 30 min linear up to 65% of B; from 30.01 to 31 min linear down to 40% of B, maintained up to 40 min. The flow rate was 1 mL/min, and using photodiode array (PDA) detection at 214 nm. Chromatographic separation was obtained with a retention time of 27.6 min, and was linear over the concentration range of 1 200 μg/mL (r2 = 0.9995). The limits of detection and quantitation were 0.34 and 1.12 μg/mL, respectively. Specificity was established in degradation studies, which also showed that there was no interference of the excipients. The accuracy was 100.22% with bias lower than 1.25%. Moreover, the in vitro cytotoxicity test of the degraded products showed non-significant differences (p>0.05). The proposed method was applied to the assessment of rhIL-11 and related proteins in biopharmaceutical dosage forms, and the results were correlated to those of a bioassay, showing a higher mean difference of the estimated content/potencies of 2.60% for the RP-LC method, aiming to establish new alternatives to monitor stability, improve quality control and thereby assure therapeutic efficacy of the biological medicine. / A interleucina 11 (IL-11) é uma citocina multifuncional que pertence a família da interleucina- 6 e estimula a proliferação, diferenciação e maturação de células hematopoiéticas. A interleucina-11 humana recombinante (rhIL-11) é produzida pela tecnologia do DNA em Escherichia coli e é usada clinicamente para a prevenção de trombocitopenia grave e redução da necessidade de transfusão de plaquetas após quimioterapia mielossupressiva em pacientes com neoplasias malignas não mielóides. No presente trabalho foi desenvolvido e validado método por cromatografia líquida em fase reversa (CL-FR) para a avaliação de rhIL-11 em formulações de produtos farmacêuticos. Utilizou-se coluna Júpiter C4 (250 mm x 4,6 mm d.i.), mantida a 25ºC. A fase móvel A foi constituída de TFA 0,1% e a fase móvel B de acetonitrila com 0,1% TFA, eluídas no seguinte gradiente: 0 0,1 min, 40% de B; 0,1 30 min, 40 65% de B; 30,01 a 31 min, 65 40% de B, mantendo-se nesta proporção até 40 min. Utilizou-se vazão de 1 mL/min e detector de arranjo de diodos (DAD) a 214 nm. A eluição cromatográfica foi obtida no tempo de 27,6 min, sendo linear na faixa de concentração de 1 200 μg/mL (r2 = 0,9995). Os limites de detecção e quantificação foram 0,34 e 1,12 μg/mL, respectivamente. A especificidade foi avaliada em estudos de degradação, que também demonstraram que não houve interferência dos excipientes. A exatidão foi 100,20%, com bias inferior a 1,25%. Além disso, realizou-se o teste de citotoxicidade in vitro das formas degradadas, as quais não apresentaram diferença significativa em relação a forma intacta (p>0,05). O método proposto foi aplicado para a avaliação da potência de rhIL-11 e de proteínas relacionadas em formulações farmacêuticas, e os resultados foram comparados com o bioensaio, observando-se diferenças das médias de conteúdo/potência 2,60% superiores para o método por CL-FR. Contribuíu-se assim para estabelecer procedimentos que aprimoram o controle da qualidade, garantindo a segurança e eficácia terapêutica do produto biotecnológico.

Febuxostat is a novel non purine drug indicated for the treatment of hyperuricemia in gout. A reversed-phase liquid chromatography (RP-LC) method was validated for the determination of febuxostat in pharmaceutical dosage forms. The LC method was carried out on a XTerra C18 column (150 mm x 3.9 mm I.D.), maintained at 25 ºC. The mobile phase consisted of water (pH 3.5) acetonitrile (40:60, v/v), run at a flow rate of 0.8 mL/min and using photodiode array (PDA) detection at 316 nm. The chromatographic separation was obtained with retention time of 3.9 min, and was linear over the range of 0.25 - 30 μg/mL (r2=0.9995). The specificity and stability-indicating capability of the method was proven through degradation studies were carried out by LC and MS and showing also, that there was no interference of the excipients and degradation products in the quantification of the drug. Moreover, the in vitro cytotoxicity test of the degraded products showed significant differences (p<0.05). The accuracy was 100.54% with bias lower than 0.65%. The limits of detection and quantitation were 0.08 and 0.28 μg/mL, respectively. The procedure was validated evaluating parameters such as the specificity, linearity, precision, accuracy, limits of detection and quantitation, robustness, and system suitability test, giving results within the acceptable range. The proposed method was applied for dissolution studies and the analysis of tablet dosage forms, contributing to assure the safety and therapeutic efficacy. / Febuxostate é um novo fármaco, não purínico, indicado para o tratamento da hiperuricemia em pacientes com gota. No presente trabalho foi desenvolvido e validado método por cromatografia líquida em fase reversa (CL-FR) para determinação de febuxostate em produtos farmacêuticos. No método por CL-FR foi utilizada coluna XTerra C18 (150 mm x 3,9 mm d.i), mantida a 25 oC. A fase móvel foi composta de água ultra-pura (pH 3,5): acetonitrila (40:60, v/v), eluída na vazão de 0,8 mL/ min com detecção no ultravioleta a 316 nm. A separação cromatográfica foi obtida no tempo de 3,9 min, sendo linear na faixa de concentração de 0,25-30 μg/mL (r2=0,9995). A especificidade do método foi comprovada através de estudos de degradação realizados por cromatografia líquida e espectrometria de massas, demonstrando que não houve interferência dos excipientes e dos produtos de degradação na quantificação do fármaco. Além disso, o teste de citotoxicidade in vitro das amostras degradadas, apresentou diferenças significativas (p <0,05) em relação à forma intacta. A precisão foi de 100,54%, com bias menor do que 0,65%. Os limites de detecção e de quantificação foram de 0,08 e 0,28 μg/mL, respectivamente. O procedimento foi validado, avaliando-se os parâmetros de especificidade, linearidade, precisão, exatidão, limite de detecção e quantificação, robustez e teste de adequabilidade do sistema, cujos resultados estão de acordo com os requisitos preconizados. O método proposto foi aplicado no estudo de dissolução e análise de formas farmacêuticas de comprimidos, contribuindo, assim, para aprimorar o controle da qualidade de medicamentos, bem como garantir a segurança e eficácia no uso terapêutico.

Nesta dissertaçao foram estudadas as reações de aminólise de benzoatos por dodecilamina (DA), propionato de dodecilamôrtio (DAP) e suas misturas em ciclohexano. Foram usados os ésteres do tipo X-&#216;-CO2-&#216;-NO2 (X=NO2,CH3O,CH3,H e Cl) e NO2-&#216;-CO2-&#216;-Y (Y=CH3 , H, Cl e CN). A aminólise de ésteres por DA é de segunda ordem em amina e está associada com entropias de ativação altamente negativas e pequenas eutalpias de ativação. A reação é muito mais sensível à natureza do grupo abandonador que da acila. Na reaçao com DAP micelar mostrou-se claramente que o grupo carboxilato de detergente atua com base geral e não como nucleófilo. A reação é de segunda ordem em DAP e apresentou as mais altas entropias de ativação. O valor de &#961; de Hammett era maior quando se variou o grupo Y, mostrando que a reação, como com DA, é mais sensível à natureza do grupo fugitivo. O valor da razão &#961;Y/&#961;X mostrou que o grupo abandonador é o íon fenóxido p-substituido. Uma prova adicional deste fato, foi obtida dos resultados de aminólise em presença de água solubilizada. O DAP micelar aumentou a velocidade de aminólise por DA, por fatôres desde 132 vêzes (Cl/NO2) até 311 vêzes (NO2/NO2 ). A aminólise de ésteres por DA+DAP apresentou parâmetros de ativação, cujos valores são intermediários àqueles para as reações com DA e com DAP respectivamente. Os valores de &#961; de Hammett foram similares aqueles obtidos para a reação com DAP somente. Baseados nos resultados previamente reportados, foram sugeridos esquemas para as reações. Eles envolvem a formação de intermediários tetraédricos (aniônicos para as reações com DA e com DAP e zwitteriônicos para a aminólise por DAP), cujo colapso é determinante da velocidade. / In this thesis the aminolysis of benzoate esters by dodecylamine (DA), dodecylammonium propionate (DAP) and their mixtures in cyclohexane was studied. The esters used were X-&#216;CO2&#216;NO2 (X=NO2CH3O,CH3,H e Cl) and NO2&#216;CO2&#216;Y (Y=CH3,H,Cl e CN). Ester aminolysis by DA is second order in the amine and is associated with a highly negative entropies and small euthalpies of activation. The reaction is much more sensitive to the nature of the leaving group than that of the acyl one. In the reaction with micellar DAP it was clearly showed that the carboxylate group of the surfact and is acting as a general base, not as a nucleophile. The reaction was found to be second order in the surfactant and showed the highest entropies of activation. The Hammett &#961; value was larger when was varied, showing that the reaction, like that with DA, is more sensitive to the nature of the leaving group. From the ratio &#961;Y/&#961;X it was shown that the leaving group is the P-substituted phenoxide ion. Additional proof for this was reached from the results of aminolysis in the presence of solubilized water. Micellar DAP enhance the rate of aminolysis by DA by factors ranging from 132 times (Cl/NO2) to 311 times (NO2/ NO2). Ester aminolysis by DA+DAP has activation parameters which are intermediate between those for the reactions with DA, and with DAP, respectively. The Hammett &#961; values were similar to those obtained for the reaction with DAP alone. Based on the previous data schemes for the reactions were suggested. They involve the formation of tetrahedral intermediates (anions for the reaction with DA and with DA+DAP, and zwitterion for the aminolysis by DAP) whose collapse are rate limiting.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The interferon is a cytokine with antiviral, antiproliferative, and immunomodulatory properties. It is a protein synthesized by cells in response to viral infection, producing successive biochemistry alterations. The chromatographic methods for evaluation of recombinant interferon-alfa 2a (rhIFN-α2a) in pharmaceutical products were validated in the present work. The reversed-phase liquid chromatography method (RP-LC) was developed and validated using a Jupiter C4 column (250 mm x 4.6 mm), maintained at ambient temperature (25°C). The mobile phase A consisted of 0.1% trifluoroacetic acid (TFA) and mobile phase B consisted of 0.1% TFA in acetonitrile, run in gradient: 0.01 1 min, 38% of B; 1 5 min, 38 43% of B; 5.01 20 min, 43 45% of B; 20.01 30 min, 45 48% of B; 30.01 40 min linear back to 38% of B and 40 42 min, 38% of B. The flow rate used was 1 mL/min with detection at 214 nm. The chromatographic separation was obtained within 42 min and it was linear in the concentration range of 0.5 50 MIU/mL (r2=0.9999). The size exclusion method was developed and validated using a BioSep-SEC-S 2000 (300 mm x 7.8 mm), maintained at ambient temperature (25°C). The mobile phase consisted of 1mM potassium phosphate monobasic, 8mM sodium phosphate dibasic and 200mM sodium chloride buffer, pH 7.4, run at a gradient flow rate: 0.01 20 min, 0.5 mL/min; 20 25 min, 0.5 1.7 mL/min; 25 35 min, 1.7 mL/min; 35 38 min, 1.7 0.5 mL/min; 38 40 min, 0.5 mL/min. The method was linear in the concentration range of 0.5 - 50 MIU/mL (r2=0.9996). The procedures were validated by the parameters of specificity, linearity, precision, accuracy, robustness, limit of quantitation and limit of detection. The methods were applied for the evaluation of the rhIFN-α2a in pharmaceutical products, contributing for the establishment of alternatives which improve the quality control, assuring the safety and therapeutic efficacy of the biological product. / O interferon é uma citocina que possui ação antiviral, imunomoduladora e antiproliferativa. É uma proteína sintetizada pelas células em resposta a infecção viral, gerando sucessivas alterações bioquímicas. No presente trabalho foram validados métodos cromatográficos para a avaliação de interferon-alfa 2a (rhIFN-α2a) em produtos farmacêuticos. O método por cromatografia líquida em fase reversa (CL-FR) foi desenvolvido e validado empregando coluna Júpiter C4 (250 mm x 4,6 mm), mantida a temperatura ambiente (25°C). A fase móvel A foi composta de ácido trifluoracético 0,1% e a fase móvel B de ácido trifluoracético 0,1% em acetonitrila, eluídas no gradiente: 0,01 1 min, 38% de B; 1 5 min, 38 43% de B; 5,01 20 min, 43 45% de B; 20,01 30 min, 45 48% de B; 30,01 40 min, 48 38% de B, mantendo-se nesta proporção até 42 min. Utilizou-se vazão de 1 mL/min e detecção no ultravioleta a 214 nm. A separação cromatográfica foi obtida no tempo de 42 min, sendo linear na faixa de concentração de 0,5 - 50 MUI/mL (r2=0,9999). Paralelamente, desenvolveu-se e validou-se método cromatográfico por exclusão molecular (CL-EM) empregando coluna BioSep-SECS 2000 (300 mm x 7,8 mm), mantida a temperatura ambiente (25°C). A fase móvel foi composta de tampão fosfato de potássio monobásico 1mM, fosfato de sódio dibásico 8mM e cloreto de sódio 200mM, pH 7,4, eluída no gradiente de fluxo: 0,01 20 min, 0,5 mL/min; 20 25 min, 0,5 1,7 mL/min; 25 35 min, 1,7 mL/min; 35 38 min, 1,7 0,5 mL/min; 38 40 min, 0,5 mL/min. O método foi linear na faixa de concentração de 0,5 - 50 MUI/mL (r2=0,9996). Ambos os procedimentos foram validados com base nos parâmetros de especificidade, linearidade, precisão, exatidão, robustez, limite de quantificação e detecção. Os métodos foram aplicados para avaliação de rhIFN-α2a em produtos farmacêuticos, contribuindo para o estabelecimento de alternativas que aprimoram o controle da qualidade, garantindo a segurança e eficácia terapêutica do produto biológico.