A indústria farmacêutica tem como principal objetivo planejar, sintetizar e caracterizar compostos químicos que possuam atividade biológica e que sejam úteis no controle e combate de doenças e sintomas que acometem as populações. Estes compostos são referidos como ingredientes farmacêuticos ativos e podem, no estado sólido, apresentar diferentes arranjos de suas moléculas dentro de um cristal (polimorfismo). A cada um desses arranjos estão associadas propriedades físico-químicas que são de fundamental importância para o efeito terapêutico dos fármacos. Nesse contexto, este estudo teve como objetivo caracterizar as propriedades de estado sólido de novas formas cristalinas de três compostos farmacêuticos, Clorpropamida (CPA), Nevirapina (NVP) e Dietilcarbamazina (DEC). A CPA trata-se de um hipoglicemiante oral, utilizado no tratamento da Diabetes mellitus tipo II, apresentando na literatura cinco polimorfos conformacionais, dos quais dois foram caracterizados nesse estudo: as Formas IV e VI. A Forma IV cristaliza-se grupo espacial não centrossimétrico monoclínico P21, com Z = 2 e a Forma VI no grupo espacial centrossimétrico ortorrômbico Pbca, com Z = 8. Ambas apresentaram o mesmo padrão de interações intermoleculares clássicas, sendo que a principal diferença entre elas reside nas interações intermoleculares não clássicas, que levam a diferentes empacotamentos cristalinos. Por fim, as conformações moleculares dos cinco polimorfos da CPA foram comparadas entre si e as informações foram racionalizadas tomando como base os resultados provenientes de cálculos teóricos, que também indicaram a possibilidade de existência de novos polimorfos. A NVP, fármaco antiretroviral não nucleosídeo utilizado no tratamento da AIDS, também apresenta casos de polimorfismo na literatura. Neste trabalho, obteve-se um solvato não estequiométrico de butanol desse composto, que cristaliza no grupo espacial centrossimétrico triclínico P-1, com Z = 2, no qual as moléculas de butanol acomodaram-se em canais infinitos rodeados por moléculas de NVP. Esse tipo de empacotamento cristalino, diferente do reportado na literatura até o presente momento, possibilitou-nos propor que novos solvatos poderiam ser obtidos variando-se o solvente, proposta tal confirmada posteriormente. A DEC é amplamente utilizada na forma de um sal de citrato no tratamento da filariose linfática, não apresentado na literatura nenhuma caracterização de estado sólido. Assim, caracterizou-se não somente o sal utilizado nas formulações (DEC citrato), como também o composto puro (DEC). A forma pura, instável à temperatura ambiente, cristaliza no grupo espacial centrossimétrico P21/n à 250K. O sal, preferido como API por sua estabilidade, cristaliza à temperatura ambiente no grupo P21/c, porém com presença de desordem nas cadeias etílicas das moléculas de DEC. Para reduzir essa desordem, efetuou-se um estudo em função da temperatura, que acabou revelando a presença de três transições de fase sólido-sólido, gerando quatro fases cristalinas diferentes. Duas das transições exibiram efeito de histerese de acordo com a direção da rampa de temperatura. A terceira transição só foi obtida por resfriamento rápido do sistema. Estes dados foram comparados com os obtidos por DSC e espectroscopia Raman / One of the main goals of the pharmaceutical industry is to plan, synthesize and characterize chemical compounds with biological activity that can be useful in controlling diseases and symptoms that affect populations. These compounds are referred to as active pharmaceutical ingredients and may, in the solid state, present different crystal arrangements of its molecules (polymorphism). To each one of these crystalline arrays are associated physicochemical properties that are of fundamental importance for the therapeutic effect of the pharmaceutical drugs. In this context, the focus of this study was to characterize the solid state properties of new crystalline forms of three pharmaceutical compounds, Chlorpropamide (CPA), Nevirapine (NVP) and Diethylcarbamazine (DEC). CPA is an oral hypoglicemiant used in the treatment of type II Diabetes mellitus, and presents five conformational polymorphs reported in the literature, two of which were characterized in this study: Forms IV and VI. Form IV crystallizes in the monoclinic non-centrosymmetric space group P21, with Z = 2 and the Form VI in the orthorrombic centrosymmetric space group Pbca, with Z = 8. Both exhibited the same classical intermolecular interaction pattern; the main difference between them lay in the non-classical intermolecular interactions, which leads to different crystal packing. Finally, the molecular conformations of the five polymorphs of CPA were compared to each other and the information was rationalized taking as basis the results from theoretical calculations, which also indicated the possible existence of new polymorphs. NVP, a non-nucleoside antiretroviral pharmaceutical compound used in the treatment of AIDS, also presents polymorphism cases reported in the literature. In this study, we obtained a non-stoichiometric buthanol solvate of this compound, which crystallizes in the triclinic centrosymmetric space group P-1, with Z = 2, in which the buthanol molecules are positioned in infinite channels surrounded by NVP molecules. This kind of crystal packing, which is different from the one reported in the literature until now, has allowed us to propose that new solvates could be obtained by varying the solvent, being this proposal subsequently confirmed. DEC is largely used as a citrate salt form in the treatment of the lymphatic filariasis, not having any solid state characterization in the literature. Thus, we characterized not just the salt (DEC citrate) used in the formulation, but also the pure compound (DEC). The pure form, unstable at room temperature, crystallizes in the monoclinic centrosymmetric space group P21/n at 250K. The salt, preferred as API because of its stability, crystallizes at room temperature in the monoclinic centrosymmetric space group P21/c, but with the presence of disorder in the ethyl chains of the DEC molecules. To reduce this disorder, we performed a study in function of temperature, which revealed the presence of three solid-solid structural phase transitions, generating four different crystalline phases. Two of these transitions showed a hysteresis effect according to the direction of the temperature ramp. The third transition was only obtained by fast cooling of the system. These data where compared with the ones obtained by DSC and Raman spectroscopy.

My dissertation focuses on (i) the development of new analysis tools and methodologies for analyzing X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) data, and (ii) the comprehensive characterization of materials (nanodiamonds) using a multi-instrument approach. Chapter 1 contains (i) a discussion of the two techniques I focused on most in my work: XPS and ToF-SIMS, (ii) a discussion of the common chemometrics techniques used to analyze data from these methods, and (iii) the advantages/rationale behind the multi-instrument characterization of materials. Chapter 2 describes various good practices for obtaining reasonable peak fits in XPS, which can also be applied to peak fitting data from different techniques. To address the issue of user subjectivity/bias in XPS peak fitting, I introduce two less biased mathematical functions for characterizing XPS narrow scans, namely the equivalent width (EW) and the autocorrelation width (AW). These functions are discussed in Chapters 3 and 4. In Chapter 5, I then introduce uniqueness plots as simple and straightforward graphical tools for assessing the quality of XPS peak fits and for determining whether fit parameters are correlated. This tool is extensively used in spectroscopic ellipsometry, and the mathematics behind it is known in XPS. However, to the best of my knowledge, this graphical tool has never been applied to XPS. ToF-SIMS data analysis is somewhat challenging due to the enormous amounts of data that are collected, and also the matrix effect in SIMS. This amount of information is significantly increased when depth profiles are performed on samples. Chapter 6 discusses a new chemometrics tool that I introduce for analysis of complex data sets, with emphasis on XPS and ToF-SIMS depth profiling data. The new approach is called the Information Content (IC) or entropy, which is adapted from Claude Shannon's work on Information Theory. Chapter 7 then contains a presentation of the comprehensive characterization of five nanodiamond samples used to manufacture particles for liquid chromatography. The advantages of a multi-instrument approach for material characterization and the lack of comprehensive material characterization in the literature are emphasized. To the best of my knowledge this is the most comprehensive characterization of nanodiamonds that has been reported in the literature. Chapter 8 presents conclusions of my work and future work. This thesis also contains six appendices. Appendix 1 contains an article from a scientific magazine that I wrote to highlight the importance and applications of the EW and AW to characterize XPS narrow scans. Appendices 2-5 are application notes I wrote on separations I performed on a nanodiamond based HPLC column. Finally, Appendix 6 describes the ToF-SIMS analysis of the tungsten species in the nanodiamond samples characterized in Chapter 7.

Middle Pennsylvanian (Desmoinesian) strata of the Lower Desert Creek (LDC) sequence within the sub-surface Greater Aneth Field (GAF) reflect a hierarchy of 4th and 5th order carbonate-dominated cycles. The Lower Desert Creek sequence, along the studied transect are composed of eight carbonate facies deposited on an east-facing shelf. There is a lateral transition from open marine algal buildup from the southeast (cores R-19, Q-16, O-16, and J-15) to a more restricted lagoonal environment to the northwest (core K-430 and E-313). The Lower Desert Creek sequence within the GAF contains three main parasequence sets: a basal, relatively deep-water unit (LDC 1), a middle skeletal to algal unit (LDC 2-4), and a shallow, open-marine/restricted lagoon unit (LDC 5-7). The southeast cores (R-19, Q-16, O-16, and J-15) contain the dolomitized basal unit in parasequence LDC 1. The northwest cores (K-430 and E-313) also contain the dolomitized basal unit in LDC 1, but show a deeper facies succession through LDC 2-4. Parasequences LDC 2-4 are the heart of the algal buildup in the GAF particularly in the southern part of the transect. The upper few parasequences (LDC 5-7) are dominated by an open marine environment represented by robust fauna. The upper parasequences (LDC 5-7) show the same shallowing upward trends with algal facies in K-430 and restricted lagoon facies in E-313. Shoaling upward trends that characterize the Lower Desert Creek sequence terminate with an exposure surface at the 4th order (Lower Desert Creek-Upper Desert Creek) sequence boundary. Porosity and permeability is weakly correlated to facies. Diagenesis within the algal reservoir is the most important factor in porosity and permeability. Marine diagenesis is observed in the form of micritization of Ivanovia, a phylloid algae. Thin fibrous isopachous rims of cloudy cement also indicate early marine diagenesis. Ghost botryoidal cements are leached during meteoric diagenesis. Meteoric drusy dog tooth cements as well as sparry calcite fill most depositional porosity. Neomorphism of micrite and the isopachous rim cements reflect meteoric diagenesis. Burial diagenesis is represented by baroque dolomite cement, compaction, and mold-filling anhydrite cement.

β-glucan is a major component of the fungal cell wall consisting of (1→3)-β linked glucose polymers with (1→6)-β linked side chains. The published classical isolation procedure of β-glucan from Saccharomyces cerevisiae is expensive and time-consuming. Thus, the aim of this research was to develop an effective procedure for the extraction of glucans. We have developed a new method for glucan extraction that will be cost effective and will maintain the native structure of the glucan. The method that we developed is 80% faster and utilizes 1/3 of the reagents compared to the published classical method. Further, the method developed increases the yield from 2.9 % to 10.3 %. Our new process has a branching frequency of 18.4 down from 197 and a side chain of 5.1 up from 2.5. The data indicate a more preserved native structure of isolated glucans.

Low-temperature heat capacity data contain information on the physical properties of materials, and new models continue to be developed to aid in the analysis and interpretation of heat capacity data into physically meaningful properties. This work presents the development of two such models and their application to real material systems. Equations describing low-energy vibrational modes with a gap in the density of states (DOS) have been derived and tested on several material systems with known gaps in the DOS, and the origins of such gaps in the DOS are presented. Lattice vacancies have been shown to produce a two-level system that can be modeled with a sum of low-energy Schottky anomalies that produce an overall linear dependence on temperature in the low-temperature heat capacity data. These two models for gaps in the vibrational DOS and the relationship between a linear heat capacity and lattice vacancies and many well-known models have been applied to several systems of materials to test their validity and applicability as well as provide greater information on the systems themselves. A series of bulk and nanoscale Mn-Fe and Co-Fe spinel solid solutions were analyzed using the entropies derived from heat capacity data, and excess entropies of mixing were determined. These entropies show that changes in valence, cation distribution, bonding, and the microstructure between the mixing ions is non-ideal, especially in the nanoparticles. The heat capacity data of ten Al doped TiO2 anatase nanoparticle samples have also been analyzed to show that the Al3+ dopant ions form small regions of short-range order, similar to a glass, within the TiO2 particles, while the overall structure of TiO2 remains unchanged. This has been supported by X-ray diffraction (XRD) and electron energy-loss spectroscopy and provides new insights to the synthesis and characterization of doped materials. The final investigation examines nanocrystalline CuO using heat capacities, magnetization, XRD, and electron microscopy and compares the findings to the known properties of bulk CuO. All of these measurements show transitions between antiferromagnetic and paramagnetic states in the temperature range of about 150-350 K that are greater in number and higher in temperature than the transitions in bulk CuO. These changes are shown to cause an increase in the temperature range of multiferroicity in CuO nanoparticles.

L’élastographie par ondes de cisaillement consiste essentiellement en deux étapes: d'abord, une onde de cisaillement est générée en appliquant soigneusement une source contrôlée externe (par exemple actionneur mécanique ou force de rayonnement ultrasonore); Alors les déplacements induits sont imagés et l'élasticité tissulaire est déduite de la propagation d'onde de cisaillement mesurée. Cependant, avec les techniques d'élastographie actuellement disponibles, l'estimation quantitative correcte de la viscosité n'est pas possible. Dans ce contexte, le but principal de cette thèse est double: d'abord, développer un outil capable d'estimer simultanément les propriétés viscoélastiques d'un solide souple, notamment en utilisant l'imagerie par cisaillement supersonique, puis de l'appliquer aux certains problèmes rencontrés en médecine et l'industrie alimentaire.La première étape consiste à utiliser la technique SSI pour générer des cartes de vitesses des ondes de cisaillement dans des milieux solides mous et isotropes. Dans ce contexte, cette technique a été utilisée pour évaluer le processus de coagulation du lait. Au cours de cette étape, les limites de la technique SSI pour obtenir une caractérisation rhéologique complète des solides viscoélastiques sont mis en évidence. Pour parvenir à cette caractérisation, la vitesse de propagation ondes de cisaillement et la absorption que l’onde subi en traversant le milieu doivent être évalués à la fois. Compte tenu des caractéristiques de la technique SSI, en raison des effets de diffraction due à la taille finie des sources, l’absorption ne peut pas être obtenue correctement de façon quantitative.La partie centrale de la thèse porte sur l’étude des avantages et des ses limites de la correction des effets de diffraction de la technique SSI à partir d’une approximation cylindrique. Cette étude a été réalisée expérimentalement en comparant les résultats obtenus avec ceux des simulations numériques. L’étude a montré que la correction cylindrique est utile pour estimer l'atténuation provoquée par l'absorption dans certaines conditions. Une fois déterminé la zone de validité de la procédure de correction, des cartes des vitesses et des cartes d'atténuation ont était obtenus. Ces cartes ont permis t la caractérisation rhéologique complète des milieux étudiés.Une fois validé cette correction nous avons procédé à réaliser trois applications d'intérêt pour obtenir les propriétés rhéologiques, une pour l’ industrie alimentaire et deux pour la médecine clinique : a) suivie et étude du processus de coagulation du lait b) suivie et étude de processus de la coagulation du sang in vitro, c) des expériences de caractérisation rhéologique du foie in vivo. Finalement, une étude de la correction cylindrique mis en œuvre a été utilisée pour caractériser rhéologiquement des solides mous avec isotropie transversale. Milieux avec différents degrés d'anisotropie ont été analysés à l'aide de simulations numériques. Les résultats obtenus montrent que pour de faibles niveaux d’anisotropie la correction cylindrique s’avère utile avec une erreur raisonnable. Pour des degrés plus élevés d'anisotropie la correction cylindrique conduit à des erreurs majeures dans l'estimation de l'atténuation. Des mesures des coefficients d’absorption ont été réalisées dans des phantoms de gel anisotropes et dans des échantillons de viande de bœuf avec des tissues musculaire / Shear wave elastography consists essentially of two steps: first, a shear wave is generated by an external controlled source (eg mechanical actuator or ultrasonic radiation force); then the induced displacements are imaged and the tissue elasticity is deduced from the measured shear wave propagation. However, with the currently available elastographic techniques, a quantitative estimation of viscosity is not possible. In this context, the objective of this thesis is twofold: first, to develop a tool capable of estimating the viscoelastic properties of a soft solid, in particular by using supersonic shear imaging (SSI), and then to apply it to some problems encountered in medicine and the food industry.The first stage consisted in using the SSI technique to generate shear wave velocity maps in soft, isotropic solid media. In this context, this technique was used to evaluate the milk coagulation process. During this stage, limitations of the SSI technique to obtain a complete rheological characterization of viscoelastic solids were demonstrated. To achieve this characterization, the velocity of the shear waves and the absorption generated when passing through the medium must be evaluated at the same time. Given the characteristics of the SSI technique, the absorption cannot be directly quantified due to diffraction.The central part of the thesis focuses in the study of the advantages and limitations of using a cylindrical approximation to correct the diffraction effects. This study was carried out experimentally and through numerical simulations. The study showed that cylindrical correction is useful for estimating the attenuation caused by absorption under certain conditions. After determining the validity zone of the correction, velocity and attenuation maps were obtained. These maps allowed the complete rheological characterization of the studied media.Once the correction was validated, three applications of interest were carried out to obtain rheological properties, one for the food industry and two for clinical medicine: a) monitoring the process of milk coagulation through storage and loss moduli maps b) following in vitro blood coagulation processes, c) rheological characterization experiments of the liver in vivo. Finally, the application of the cylindrical correction to characterize transversely isotropic soft solids was studied. Media with different degrees of anisotropy were analyzed through numerical simulations. The results obtained showed that for low levels of anisotropy the cylindrical correction proves useful within a reasonable error. For higher degrees of anisotropy the cylindrical correction leads to major errors in the estimation of the attenuation. Measurements of the absorption coefficients were performed in anisotropic gel phantoms and in beef samples with muscle tissue

Lignin, the second most abundant naturally occurring polymer found in plant cell wall has the potential of becoming an alternative source for the production of chemical synthons for the pharmaceuticals and other chemical industries. While much gain has been made towards the development of degradation methods to break down lignin, effective analytical methods are still required to rapidly and accurately identify the products of lignin breakdown experiments. The goal of this work was to develop mass spectrometric methods for the characterization of lignin oligomers based on the study of model lignin compounds. Unlike peptides and oligosaccharides, lignin model compounds that could serve as analytical standards for methods developments are not commercially available, and hence, the first project of this dissertation focused on the synthesis of lignin model compounds containing the β-O-4 bond. The priority was to synthesize compounds containing all important functionalities that reflect the structure of native lignin. By employing the known Aldol reaction, lignin oligomers containing the β-O-4 were synthesized. The synthesized β-O-4 lignin oligomers contained the characteristic functional groups of native β-O-4 lignin, that is, the phenolic functionality, the aryl glycerol β-O-4 aryl ether bond and the unsaturated side chain. The second project was aimed at developing alternative ionization methods for the characterization of lignin in the negative ion mode mass spectrometry. A chloride adduct ionization method was developed and used for characterizing and sequencing lignin oligomers. This method proved to be very useful in stabilizing the adduct ion in the full scan spectrum mode and also providing useful structural information upon tandem mass spectrometry. In the third project, a cationization technique was developed to unambiguously assign the sequence in which β-O-4 lignin oligomers are connected. A simple and easy to use sequencing chart was designed and could serve as a guide for predicting the sequence of larger lignin oligomers. This method offers an alternative approach for the characterization of lignin oligomers in the positive ion mode mass spectrometry. The fourth project focused on the ionization response of a new class of β-O-4 lignin compounds. β-O-4 compounds having the same skeletal backbone but different non-polar groups at the a-position were synthesized, and their ESI response studied. Results from this study show that a slight change in the structure of a β-O-4 lignin model compound can change the cationization response to several order of magnitude. Most importantly, this work for the first time has shown a direct correlation between lignin ionization response and lignin structure. The fifth project was aimed at studying the chromatographic behavior of the diastereomer pair in β-O-4 lignin model compounds. Using three commercially available HPLC columns, the chromatographic behavior and factors that affect the separation of the diastereomer pair of the β-O-4 lignin diastereomer on an HPLC column were studied. By performing tandem mass spectrometry on each of the diastereomers, a fragmentation mechanism was developed that could be used to unambiguous assign the configuration (erythro or threo) for the pair of diastereomer in a β-O-4 model. The results presented in this dissertation adds significant knowledge to the lignin mass spectrometry literature, and it offers new ionization techniques for the characterization of lignin oligomers, most importantly, an alternative approach for lignin analysis using adduct ionization mass spectrometry. The developed methods could easily be extended for the characterization of larger lignin oligomers.

Lactic acid bacteria have been reported to be used as a health adjunct in food for u many years. However, these health benefits have not been proven. and how these bacteria pass through the digestion process and remain viable in the human intestinal tract is still not clear. The aim of this work was to isolate mutants from Lactobacillus acidophilus or Bifidobacterium bifidum that could tolerate the conditions of the digestion process (low pH and bile conduction) and to characterize these isolated mutants. Acid- and bile-tolerant mutants of L. acidophilus were isolated from parental strains successfully using natural selection techniques. These mutants survived and grew at conditions of pH 3.5 with 0.2% mixed bile salts added. After the selection, phenotypic characterization was identified to further clarify desirable traits for use as probiotic adjuncts in foods. These phenotypic characteristics included protease, aminopeptidase, ß-galactosidase, and bile salt hydrolase activity. Based on different protease, aminopeptidase, and ß-galactosidase activity, selected acid- and bile-tolerant mutants contained different growth characteristics compared with their parents. All the isolates tested showed different bile salt hydrolase activity, and this activity was not strain and medium dependent. Plasmid profiles and fatty acid analysis were conducted to provide more information of these acid/bile tolerant isolates and whether or not they were mutants from their parent strains rather than only adapted variants. Results showed the acid-/bile-tolerant isolates contained different plasmid profiles and cell wall fatty acids compared with their parents, which indicated these isolates were mutants. Protein expression by two-dimensional gel electrophoresis showed different protein expression patterns between acid- and bile-tolerant mutants and their parents. fm1her suggesting these isolates were mutants. We observed the protein production in parent strains decreased as the pH decreased. and protein expression in mutants remained the same as pH decreased. Two of the proposed health benefits of probiotic bacteria are anticholesterol activity and antimicrobial activity. These were evaluated using selected acid- and bile-tolerant mutants. Results showed no decrease of cholesterol in the test medium during bacterial growth. The observed antimicrobial activity was due to the presence of active cells. and this may relate to the acid production during cell growth and not to the production of antimicrobial substances. We concluded that the acid-/bile-tolerant isolates were mutants, and they survived and grew better in harsh environments compared with their parent strains. These mutants may be useful as a food adjunct in the future, but further study is needed to establish their use and possible probiotic benefits.

This study represents an effort to interrupt the life cycle of Eimeria tenella, the parasite that causes cecal coccidiosis in chickens, with a panel of 12 gamont-specific monoclonal antibodies (Mabs). To facilitate Mab screening, it was first necessary to develop a cell culture-adapted strain of E. tenella (field strain 80) from primary chicken kidney cells (PCKC) to white Leghorn chickens and back to PCKC. This alternation was repeated through 12 such passages. As a result, we have developed a cell culture-adapted strain of E. tenella that produces over 280% more oocysts in vitro than the original parent strain, and over six times as many oocysts as reported by other investigators. Of the 12 Mabs evaluated, Mab HD8 had the greatest inhibitory effect by significantly reducing (P<0.05) the number of E. tenella oocysts produced in cell culture by 61% compared with untreated controls. Western blot analysis revealed that the target antigen for Mab HD8 had a molecular mass of 71 and 36 kilodaltons, respectively, when run under nonreduced and reduced conditions. Subsequent evaluation of soluble and membrane-associated proteins indicated that the target antigen for Mab HD8 was membrane-associated. In vivo studies are now required to validate the efficacy of this antigen in a subunit vaccine.

High entropy alloys (HEA) are a relatively new group of alloys first introduced in 2004. They usually contain 5 to 6 different principle elements. Each of these elements comprise 5-35 at. % of the chemical composition of the alloy. There is a growing interest in the research community about the development of these alloys as well as their engineering applications. Some HEAs have interesting properties that have made them well suited for higher temperature applications, particularly refractory uses, while some have been shown to maintain their mechanical properties even at cryogenic temperatures. Initially, the HEA research was focused on developing alloys with equiatomic compositions as it was believed that the single phase HEA would only form at such composition ratios. However, further research have found multiple HEAs with non-equiatomic chemical compositions. A major question that needs to be answered at this point is how to identify these non-equiatomic single phase alloy systems. Unlike the conventional alloys, the HEAs do not have a base element as a solvent, which complicates the identification of new alloy systems via conventional development techniques. To find a potential HEA, alloy development techniques of both exploratory and computational natures are being conducted within the community. Even though multiple HEAs have been successfully identified and fabricated by these techniques, in most cases they require extensive experimental data and are relatively time consuming and expensive. This study proposes a thin film combinatorial approach as a more efficient experimental method in developing new HEA alloy systems. In order to study HEA systems with different crystal structures, nominal HEA compositions were selected, including: CoFeMnNiCu in order to achieve face centered cubic (FCC) HEA, OsRuWMoRe to obtain hexagonal closed packed (HCP) and VNbMoTaW in an attempt to form a body centered cubic (BCC) crystal structure. Thin film samples were fabricated by simultaneous magnetron sputtering of the elements onto silicon wafer substrates. The arrangement of the sputtering targets yielded a chemical composition gradient in the films which ultimately resulted in the formation of various phases. Some of these phases exhibited the desired single-phase HEA, albeit with non-equiatomic chemical compositions. Bulk samples of the identified HEA compositions were prepared by arc melting mixtures of the metals. Microstructure of both thin film samples and bulk samples were characterized via scanning electron microscopy (SEM), focused ion beam (FIB) and energy dispersive x-ray spectroscopy (EDX). The crystal structures of the samples were studied by X-ray diffraction (XRD) and electron backscattered diffraction (EBSD) technique. Applying nano-indentation technique, the mechanical properties of some of the samples were screened over the composition gradient as well. By applying this combinatorial thin film approach, single-phase FCC, HCP and BCC HEAs were detected and successfully produced in bulk form. Additionally, screening the properties of the compositionally gradient thin films, as well as their chemical composition and crystal structure, provided a thorough understanding of the phase space. This experimental approach proved to be more efficient in identifying new alloy systems than conventional exploratory development methods.