Este projeto de pesquisa, com o apoio dos pequenos produtores de café do Estado de São Paulo/Brasil, visou estudar e caracterizar o óleo de grãos de café verde obtido pela extração supercrítica e extração com fluido pressurizado. Foi feita a caracterização deste produto, considerando os componentes minoritários com atividade biológica demonstrada na literatura, como os componentes fenólicos e os diterpenos cafestol e kahweol. Com o objetivo de obter o óleo de café verde enriquecido, principalmente nestes dois principais diterpenos, componentes relevantes para as indústrias cosmética e farmacêutica, aplicaram-se como inovação tecnológica a extração supercrítica (SFE) e extração com fluido pressurizado (PFE). Os ensaios de extração foram realizados no Laboratório a Alta Pressão e Produtos Naturais (FZEA/USP/Brasil) para as SFE e no Laboratoire de Procédés (IUT/Université d\'Orléans/França) para PFE. No desenvolvimento da metodologia de análise dos diterpenos foram realizados estudos do método de saponificação do óleo de café verde Arabica (Coffea arabica) cv. Catuaí amarelo, obtido por prensagem (OP). E, concomitantemente a este estudo analítico, foram feitas as extrações do óleo dos mesmos grãos de café verde via SFE e PFE. Os diferentes óleos de grãos de café verde, obtidos pelos três métodos de extração foram analisados e comparados entre si considerando principalmente a concentração dos diterpenos. A metodologia analítica desenvolvida na primeira fase do trabalho constatou que para uma melhor análise dos diterpenos a reação de saponificação deve ser feita utilizando NaOH e a 70 °C. A fração insaponificável foi analisada para os diferentes extratos. Para os extratos obtidos via SFE, a cafeína teve que ser separada do óleo. A análise de quantificação dos diterpenos via cromatografia gasosa, utilizando o próprio cafestol e kahweol como padrão externo, indicou que o óleo obtido por SFE apresentou 50.180,41 mg cafestol/kg óleo e 63.764,32 mg kahweol/kg óleo na condição otimizada. Para a PFE as concentrações máximas obtidas foram de 65.876,29 mg cafestol/kg óleo e 53.328,47 mg kahweol/kg óleo e, no óleo de café verde obtido pelo método convencional a concentração dos diterpenos mostrou-se inferior com 7.508 mg cafestol/kg óleo e 12.822 mg kahweol/kg óleo. As propriedades físicas medidas visando a caracterização do óleo dos grãos de café verde obtido por SFE e prensagem dos grãos indicaram que tanto a densidade quanto a viscosidade diminuem com o aumento da temperatura. Para ambos os óleos, os comportamento da densidade foi semelhante, para aquele obtido pelo método convencional, os valores variaram de 0,94177 g/cm3 (25 °C) a 0,92143 g/cm3 (55 °C) e para o óleo obtido por SFE a variação foi de 0,93645 g/cm3 (25 °C) a 0,9157 g/cm3 (55 °C). A viscosidade dinâmica para o óleo obtido por prensagem variou de 127,8798 mPa×s (25 °C) a 35,0510 mPa×s (55 °C) e, para o óleo obtido por SFE, estes valores foram menores, entre 84,0411 mPa×s (25 °C) e 24,2555 mPa×s (55 °C). O índice de refração para ambos os óleos foi semelhante. Na extração com fluido pressurizado, (PFE), um valor ótimo de rendimento do extrato foi obtido quando se empregou metanol como solvente para uma temperatura de 100 °C e tempo de equilíbrio estático de 20 min. Este valor foi 183% superior a media dos resultados obtidos nos outros ensaios. Na concentração de diterpenos nestes extratos, nota-se também que o etanol foi menos eficiente que o metanol. Na comparação dos dois processos inovadores (SFE e PFE), a concentração de kahweol apresenta-se como o principal componente em quase todos os ensaios, com valores da relação cafestol/kahweol entre 0,73 e 0,96. No entanto, ao ser analisado o processo individualmente, percebe-se que a extração com fluido supercrítico indica que a maioria dos extratos obtidos teve uma concentração de kahweol superior ao cafestol. / This research project, with the support of small coffee producers in the State of São Paulo/Brazil, aimed to study and characterize the green coffee oil obtained by the supercritical fluid extraction and pressurized fluid extraction. Was made to characterize this authentic product, considering the minor components with biological activity demonstrated in the literature, such as phenolic compounds and diterpenes cafestol and kahweol. In order to get the green coffee oil enriched, mainly in these two major diterpenos, components relevant to cosmetic and pharmaceutical industries, applied technological innovation as the supercritical fluid extraction (SFE) and pressurized fluid extraction (PFE). The extraction tests were performed at the High Pressure Laboratory and Natural Products (FZEA/USP/Brazil) for SFE and the Laboratoire de Procédés (IUT/Université d\' Orléans/France) for PFE. In the development of methodology for analyzing diterpenes were conducted studies on the method of saponification of the green oil Arabica coffee (Coffea arabica) cv. yellow Catuaí, obtained by pressing (OP). And, concomitantly this analytical study were extracted from the oil of the same green coffee beans by SFE and PFE. Different green coffee beans oils, obtained by the three extraction methods were analyzed and compared with each other considering mainly the concentration of diterpenes. The analytical methodology developed in the first phase of the work found that for a better analysis of the diterpenes saponification reaction must be made using NaOH and 70 °C. The unsaponifiable fraction was analyzed for the different extracts. Specifically for extracts obtained by SFE, caffeine had to be separated from the oil. The analysis and quantification of diterpenes made by gas chromatography, using himself as cafestol and kahweol external standard, indicated that the oil obtained by SFE showed cafestol 50,180.41 mg/kg oil and kahweol 63,764.32 mg/kg oil in optimal condition. For the PFE maximum concentrations were obtained cafestol 65,876.29 mg/kg oil and kahweol 53,328.47 mg/kg oil and the green coffee oil obtained by the conventional method the concentration of diterpenes was lower than with 7,508 mg cafestol/kg oil kahweol and 12,822 mg/kg oil. The physical measures for the characterization of oil from green coffee beans obtained by SFE and pressing the grains indicated that both the density and the viscosity decrease with increasing temperature. For both oils, the density behavior was similar, to that obtained by the conventional method, the values ranged from 0.94177 g/cm3 (25 °C) to 0.92143 g/cm3 (55 °C) and the oil obtained by the SFE variation was 0.93645 g/cm3 (25 °C) to 0.9157 g/cm3 (55 °C). The dynamic viscosity for the oil obtained by pressing ranged from 127.8798 mPa×s (25 °C) to 35.0510 mPa×s (55 °C) and the oil obtained by SFE, these values were lower, between 84.0411 mPa×s (25 °C) and 24.2555 mPa×s (55 °C). The index of refraction for both oils was similar. In the extraction pressure, (PFE), an optimal value of the extract yield was obtained when methanol was used as solvent at a temperature of 100 °C and static equilibrium time of 20 min. This value was 183% above the average of the results obtained in other tests. At the concentration of these diterpenes extracts, note also that the ethanol was less effective than methanol. In comparing the two innovative processes (SFE and PFE), the concentration of kahweol presents itself as the main component in almost all tests, compared with values of cafestol/kahweol between 0.73 and 0.96. However, the process to be analyzed individually, it is clear that the supercritical fluid extraction indicates that most of the extracts had a concentration higher of kahweol than cafestol.

Por meio do método lattice Boltzmann (LBM), este projeto de pesquisa propõe a simulação computacional da extração dos compostos diterpênicos do café verde em sistemas com fluido pressurizado. Para isso foi adotado um modelo fenomenológico dinâmico e de primeira ordem quanto à dependência espacial. Usando a rede D1Q2, o LBM foi implementado com duas funções de distribuição de partículas, acompanhadas das respectivas funções de distribuição de equilíbrio. Cada função refere-se à concentração em uma dada fase (sólida e fluida). A previsão das curvas de rendimento pressupõe o conhecimento dos parâmetros do processo, tais como a difusividade mássica interna (intrapartícula), o coeficiente de partição volumétrica e a difusividade mássica axial. Para o devido ajuste destes parâmetros contra dados experimentais, foi desenvolvido um código computacional utilizando a técnica de gradiente descendente. Resultados numéricos das simulações foram comparados com dados experimentais e mostraram-se satisfatórios. Adicionalmente, foi realizada a análise da influência dos parâmetros supracitados mediante a criação de uma interface gráfica amigável para o código computacional original. Este projeto de pesquisa, além de fomentar a agregação de valor ao café, desenvolveu competências em modelagem e simulação de processos de extração de espécies químicas a partir de métodos numéricos. / Using the lattice Boltzmann method (LBM), this research project proposes a computer simulation of diterpene compounds extraction from green coffee beans in high pressure fluid systems. In order to do so, a dynamic phenomenological model with first-order spatial dependence was adopted. LBM was implemented on D1Q2 lattice, using two particle distribution functions, followed by the respective equilibrium distribution functions. Each function refers to the extract concentration in each phase, namely solid and fluid. Prediction of extract yield curves relies on process parameters such as internal (i.e., intraparticle) mass diffusivity, volumetric partition coefficient and axial mass diffusivity. Values of aforesaid parameters were fitted against experimental data via descendent gradient technique coupled to the LBM computer code and numerically simulated result were consistent. Besides modeling and simulation, this project performed the influence analysis of those process parameters. To this end, a user-friendly graphical interface for the original computer code was created. While fostering value to be added to coffee, this research project develops skills in modeling and simulation of chemical species extraction through numerical methods.

Por meio do método lattice Boltzmann (LBM), este projeto de pesquisa propõe a simulação computacional da extração dos compostos diterpênicos do café verde em sistemas com fluido pressurizado. Para isso foi adotado um modelo fenomenológico dinâmico e de primeira ordem quanto à dependência espacial. Usando a rede D1Q2, o LBM foi implementado com duas funções de distribuição de partículas, acompanhadas das respectivas funções de distribuição de equilíbrio. Cada função refere-se à concentração em uma dada fase (sólida e fluida). A previsão das curvas de rendimento pressupõe o conhecimento dos parâmetros do processo, tais como a difusividade mássica interna (intrapartícula), o coeficiente de partição volumétrica e a difusividade mássica axial. Para o devido ajuste destes parâmetros contra dados experimentais, foi desenvolvido um código computacional utilizando a técnica de gradiente descendente. Resultados numéricos das simulações foram comparados com dados experimentais e mostraram-se satisfatórios. Adicionalmente, foi realizada a análise da influência dos parâmetros supracitados mediante a criação de uma interface gráfica amigável para o código computacional original. Este projeto de pesquisa, além de fomentar a agregação de valor ao café, desenvolveu competências em modelagem e simulação de processos de extração de espécies químicas a partir de métodos numéricos. / Using the lattice Boltzmann method (LBM), this research project proposes a computer simulation of diterpene compounds extraction from green coffee beans in high pressure fluid systems. In order to do so, a dynamic phenomenological model with first-order spatial dependence was adopted. LBM was implemented on D1Q2 lattice, using two particle distribution functions, followed by the respective equilibrium distribution functions. Each function refers to the extract concentration in each phase, namely solid and fluid. Prediction of extract yield curves relies on process parameters such as internal (i.e., intraparticle) mass diffusivity, volumetric partition coefficient and axial mass diffusivity. Values of aforesaid parameters were fitted against experimental data via descendent gradient technique coupled to the LBM computer code and numerically simulated result were consistent. Besides modeling and simulation, this project performed the influence analysis of those process parameters. To this end, a user-friendly graphical interface for the original computer code was created. While fostering value to be added to coffee, this research project develops skills in modeling and simulation of chemical species extraction through numerical methods.

<p>Different sample preparation steps prior to a detection method are often essential in analytical chemistry. In this thesis, both static extractions and on-line coupled solid-phase extractions have been studied in combination with different detection techniques. Aspects of performing sample preparations in miniaturized analytical devices and the development of poly(dimethylsiloxane) (PDMS) microchips are discussed. Polypyrrole was also evaluated as an electrochemically controllable stationary phase for solid-phase microextraction (SPME) and solid-phase extraction (SPE).</p><p>The first part of this thesis describes the extraction of an organic compound from a very complex solid matrix utilizing the pressurized-fluid extraction (PFE) technique. The presented results show that PFE is easily optimized and enables rapid extractions and extracts relatively free from interferences.</p><p>An integrated three-electrode device, which enabled electrochemical (EC) SPME under potential control, was developed. With this device, both anions and cations could be extracted employing two types of polypyrrole films. Planar micro band electrodes positioned at the end of a capillary were also used to electrochemically extract and detect anions in a miniaturized flow system. Different analyte concentrations and preconcentration times were examined, and good linear correlations were found between the extraction time and the detection response. The on-line coupling of a thin layer EC cell, with a polypyrrole coated working electrode, to different mass spectrometric (MS) techniques is also described and evaluated. The results show that EC-SPE, employing polypyrrole as stationary phase, can be used as a preconcentration step prior to detection.</p><p>In addition, this thesis describes the development and on-line coupling of a microelectrode array equipped PDMS microchip with an integrated graphite electrospray emitter to electrospray ionization (ESI) MS. The system enabled short transfer times and an EC conversion efficiency of 30% at a flow rate of 0.5 μL/min. The on-line EC/ESI-MS experiments were significantly simplified using a wireless Bluetooth battery-powered EC instrument.</p>

<p>This thesis covers a wide range of analytical method development for lipid analysis in complex biological samples; from sample preparation using pressurized fluid extraction (PFE) and separation with reversed phase capillary liquid chromatography (RP-LC) to detection by electrospray ionization mass spectrometry (ESI/MS) and tandem MS.</p><p>The requirements for fast, reliable and selective extraction methods with minimal usage of solvents have accelerated the development of new extraction techniques. PFE is one of the new automated, fast and efficient liquid extraction techniques which use elevated temperature and pressure with standard liquid solvents. In this thesis the reliability and efficiency of the PFE technique was investigated for the extraction of total lipid content from cod, herring muscle and human brain tissue as well as for pesticides from fatty foodstuffs. Improved or comparable efficiencies were achieved with reduced time and solvent consumption as compared to traditional methods. </p><p>A RP-LC coupled online to ESI/MS for the analysis of phosphatidylcholine (PC) and sphingomyelin (SM) molecular species was developed and used for the analysis of brain lipids from eight groups of mice treated with vehicle and various neuroleptics. The effect of postnatal iron administration in lipid composition and behavior was investigated. Whether or not these effects could be altered by subchronic administration of the neuroleptics (clozapine and haloperidol) were examined. The results support the hypothesis that an association between psychiatric disorders, behavior abnormalities and lipid membrane constitution in the brain exists.</p><p>Finally, a tandem MS precursor ion scan was used to analyze the developmental profile of brain sulfatide accumulation in arylsulfatase A (ASA) deficient (ASA -/-) as compared to wild type control (ASA +/+) mice. The ASA -/- mice were developed as a model of the monogenic disease metachromatic leukodystrophy with an established deficiency of the lysosomal enzyme ASA. The results showed that an alteration in the composition of sulfatide molecular species was observed between the ASA -/- and ASA +/+ mice.</p><p>This thesis shows that modern analytical methods can provide new insights in the extraction and analysis of lipids from complex biological samples.</p>

<p>The challenge for the future was defined by the Brundtland Commission (1987) and by the Rio Declaration (1992), in which the fundamental principles for achieving a sustainable development were provided. Sustainable chemistry can be defined as the contribution of chemistry to the implementation of the Rio Declaration. This thesis shows how Pressurized Fluid Extraction (PFE) can be utilized in chemical analysis, and how this correlates to Green Chemistry.</p><p>The reliability and efficiency of the PFE technique was investigated for a variety of analytes and matrices. Applications discussed include: the extraction of the antioxidant Irganox 1076 from linear low density polyethylene, mobile forms of phosphorus in lake sediment, chlorinated paraffins from source-separated household waste, general analytical method for pesticide residues in rape seed, total lipid content in cod muscle, and squalene in olive biomass. Improved or comparable extraction yields were achieved with reduced time and solvent consumption. The decrease in use of organic solvents was 50-90%, resulting in minimal volatile organic compounds emissions and less health-work problem. Due to higher extraction temperatures and more efficient extractions, the selection of solvent is not as important as at lower temperatures, which makes it possible to choose less costly, more environmentally and health beneficial solvents. In general, extraction times are reduced to minutes compared to several hours. As a result of the very short extraction times, the amount of co-extracted material is relatively low, resulting in fewer clean-up step and much shorter analysis time. Selective extractions could be obtained by varying the solvent or solvent mixture and/or using adsorbents. </p><p>In this thesis, the PFE technique was compared to the twelve principles of Green Chemistry, and it was shown that it follows several of the principles, thus giving a major contribution to sustainable chemistry. </p>

Different sample preparation steps prior to a detection method are often essential in analytical chemistry. In this thesis, both static extractions and on-line coupled solid-phase extractions have been studied in combination with different detection techniques. Aspects of performing sample preparations in miniaturized analytical devices and the development of poly(dimethylsiloxane) (PDMS) microchips are discussed. Polypyrrole was also evaluated as an electrochemically controllable stationary phase for solid-phase microextraction (SPME) and solid-phase extraction (SPE). The first part of this thesis describes the extraction of an organic compound from a very complex solid matrix utilizing the pressurized-fluid extraction (PFE) technique. The presented results show that PFE is easily optimized and enables rapid extractions and extracts relatively free from interferences. An integrated three-electrode device, which enabled electrochemical (EC) SPME under potential control, was developed. With this device, both anions and cations could be extracted employing two types of polypyrrole films. Planar micro band electrodes positioned at the end of a capillary were also used to electrochemically extract and detect anions in a miniaturized flow system. Different analyte concentrations and preconcentration times were examined, and good linear correlations were found between the extraction time and the detection response. The on-line coupling of a thin layer EC cell, with a polypyrrole coated working electrode, to different mass spectrometric (MS) techniques is also described and evaluated. The results show that EC-SPE, employing polypyrrole as stationary phase, can be used as a preconcentration step prior to detection. In addition, this thesis describes the development and on-line coupling of a microelectrode array equipped PDMS microchip with an integrated graphite electrospray emitter to electrospray ionization (ESI) MS. The system enabled short transfer times and an EC conversion efficiency of 30% at a flow rate of 0.5 μL/min. The on-line EC/ESI-MS experiments were significantly simplified using a wireless Bluetooth battery-powered EC instrument.

This thesis covers a wide range of analytical method development for lipid analysis in complex biological samples; from sample preparation using pressurized fluid extraction (PFE) and separation with reversed phase capillary liquid chromatography (RP-LC) to detection by electrospray ionization mass spectrometry (ESI/MS) and tandem MS. The requirements for fast, reliable and selective extraction methods with minimal usage of solvents have accelerated the development of new extraction techniques. PFE is one of the new automated, fast and efficient liquid extraction techniques which use elevated temperature and pressure with standard liquid solvents. In this thesis the reliability and efficiency of the PFE technique was investigated for the extraction of total lipid content from cod, herring muscle and human brain tissue as well as for pesticides from fatty foodstuffs. Improved or comparable efficiencies were achieved with reduced time and solvent consumption as compared to traditional methods. A RP-LC coupled online to ESI/MS for the analysis of phosphatidylcholine (PC) and sphingomyelin (SM) molecular species was developed and used for the analysis of brain lipids from eight groups of mice treated with vehicle and various neuroleptics. The effect of postnatal iron administration in lipid composition and behavior was investigated. Whether or not these effects could be altered by subchronic administration of the neuroleptics (clozapine and haloperidol) were examined. The results support the hypothesis that an association between psychiatric disorders, behavior abnormalities and lipid membrane constitution in the brain exists. Finally, a tandem MS precursor ion scan was used to analyze the developmental profile of brain sulfatide accumulation in arylsulfatase A (ASA) deficient (ASA -/-) as compared to wild type control (ASA +/+) mice. The ASA -/- mice were developed as a model of the monogenic disease metachromatic leukodystrophy with an established deficiency of the lysosomal enzyme ASA. The results showed that an alteration in the composition of sulfatide molecular species was observed between the ASA -/- and ASA +/+ mice. This thesis shows that modern analytical methods can provide new insights in the extraction and analysis of lipids from complex biological samples.

The challenge for the future was defined by the Brundtland Commission (1987) and by the Rio Declaration (1992), in which the fundamental principles for achieving a sustainable development were provided. Sustainable chemistry can be defined as the contribution of chemistry to the implementation of the Rio Declaration. This thesis shows how Pressurized Fluid Extraction (PFE) can be utilized in chemical analysis, and how this correlates to Green Chemistry. The reliability and efficiency of the PFE technique was investigated for a variety of analytes and matrices. Applications discussed include: the extraction of the antioxidant Irganox 1076 from linear low density polyethylene, mobile forms of phosphorus in lake sediment, chlorinated paraffins from source-separated household waste, general analytical method for pesticide residues in rape seed, total lipid content in cod muscle, and squalene in olive biomass. Improved or comparable extraction yields were achieved with reduced time and solvent consumption. The decrease in use of organic solvents was 50-90%, resulting in minimal volatile organic compounds emissions and less health-work problem. Due to higher extraction temperatures and more efficient extractions, the selection of solvent is not as important as at lower temperatures, which makes it possible to choose less costly, more environmentally and health beneficial solvents. In general, extraction times are reduced to minutes compared to several hours. As a result of the very short extraction times, the amount of co-extracted material is relatively low, resulting in fewer clean-up step and much shorter analysis time. Selective extractions could be obtained by varying the solvent or solvent mixture and/or using adsorbents. In this thesis, the PFE technique was compared to the twelve principles of Green Chemistry, and it was shown that it follows several of the principles, thus giving a major contribution to sustainable chemistry.

Este projeto de pesquisa, com o apoio dos pequenos produtores de café do Estado de São Paulo/Brasil, visou estudar e caracterizar o óleo de grãos de café verde obtido pela extração supercrítica e extração com fluido pressurizado. Foi feita a caracterização deste produto, considerando os componentes minoritários com atividade biológica demonstrada na literatura, como os componentes fenólicos e os diterpenos cafestol e kahweol. Com o objetivo de obter o óleo de café verde enriquecido, principalmente nestes dois principais diterpenos, componentes relevantes para as indústrias cosmética e farmacêutica, aplicaram-se como inovação tecnológica a extração supercrítica (SFE) e extração com fluido pressurizado (PFE). Os ensaios de extração foram realizados no Laboratório a Alta Pressão e Produtos Naturais (FZEA/USP/Brasil) para as SFE e no Laboratoire de Procédés (IUT/Université d\'Orléans/França) para PFE. No desenvolvimento da metodologia de análise dos diterpenos foram realizados estudos do método de saponificação do óleo de café verde Arabica (Coffea arabica) cv. Catuaí amarelo, obtido por prensagem (OP). E, concomitantemente a este estudo analítico, foram feitas as extrações do óleo dos mesmos grãos de café verde via SFE e PFE. Os diferentes óleos de grãos de café verde, obtidos pelos três métodos de extração foram analisados e comparados entre si considerando principalmente a concentração dos diterpenos. A metodologia analítica desenvolvida na primeira fase do trabalho constatou que para uma melhor análise dos diterpenos a reação de saponificação deve ser feita utilizando NaOH e a 70 °C. A fração insaponificável foi analisada para os diferentes extratos. Para os extratos obtidos via SFE, a cafeína teve que ser separada do óleo. A análise de quantificação dos diterpenos via cromatografia gasosa, utilizando o próprio cafestol e kahweol como padrão externo, indicou que o óleo obtido por SFE apresentou 50.180,41 mg cafestol/kg óleo e 63.764,32 mg kahweol/kg óleo na condição otimizada. Para a PFE as concentrações máximas obtidas foram de 65.876,29 mg cafestol/kg óleo e 53.328,47 mg kahweol/kg óleo e, no óleo de café verde obtido pelo método convencional a concentração dos diterpenos mostrou-se inferior com 7.508 mg cafestol/kg óleo e 12.822 mg kahweol/kg óleo. As propriedades físicas medidas visando a caracterização do óleo dos grãos de café verde obtido por SFE e prensagem dos grãos indicaram que tanto a densidade quanto a viscosidade diminuem com o aumento da temperatura. Para ambos os óleos, os comportamento da densidade foi semelhante, para aquele obtido pelo método convencional, os valores variaram de 0,94177 g/cm3 (25 °C) a 0,92143 g/cm3 (55 °C) e para o óleo obtido por SFE a variação foi de 0,93645 g/cm3 (25 °C) a 0,9157 g/cm3 (55 °C). A viscosidade dinâmica para o óleo obtido por prensagem variou de 127,8798 mPa×s (25 °C) a 35,0510 mPa×s (55 °C) e, para o óleo obtido por SFE, estes valores foram menores, entre 84,0411 mPa×s (25 °C) e 24,2555 mPa×s (55 °C). O índice de refração para ambos os óleos foi semelhante. Na extração com fluido pressurizado, (PFE), um valor ótimo de rendimento do extrato foi obtido quando se empregou metanol como solvente para uma temperatura de 100 °C e tempo de equilíbrio estático de 20 min. Este valor foi 183% superior a media dos resultados obtidos nos outros ensaios. Na concentração de diterpenos nestes extratos, nota-se também que o etanol foi menos eficiente que o metanol. Na comparação dos dois processos inovadores (SFE e PFE), a concentração de kahweol apresenta-se como o principal componente em quase todos os ensaios, com valores da relação cafestol/kahweol entre 0,73 e 0,96. No entanto, ao ser analisado o processo individualmente, percebe-se que a extração com fluido supercrítico indica que a maioria dos extratos obtidos teve uma concentração de kahweol superior ao cafestol. / This research project, with the support of small coffee producers in the State of São Paulo/Brazil, aimed to study and characterize the green coffee oil obtained by the supercritical fluid extraction and pressurized fluid extraction. Was made to characterize this authentic product, considering the minor components with biological activity demonstrated in the literature, such as phenolic compounds and diterpenes cafestol and kahweol. In order to get the green coffee oil enriched, mainly in these two major diterpenos, components relevant to cosmetic and pharmaceutical industries, applied technological innovation as the supercritical fluid extraction (SFE) and pressurized fluid extraction (PFE). The extraction tests were performed at the High Pressure Laboratory and Natural Products (FZEA/USP/Brazil) for SFE and the Laboratoire de Procédés (IUT/Université d\' Orléans/France) for PFE. In the development of methodology for analyzing diterpenes were conducted studies on the method of saponification of the green oil Arabica coffee (Coffea arabica) cv. yellow Catuaí, obtained by pressing (OP). And, concomitantly this analytical study were extracted from the oil of the same green coffee beans by SFE and PFE. Different green coffee beans oils, obtained by the three extraction methods were analyzed and compared with each other considering mainly the concentration of diterpenes. The analytical methodology developed in the first phase of the work found that for a better analysis of the diterpenes saponification reaction must be made using NaOH and 70 °C. The unsaponifiable fraction was analyzed for the different extracts. Specifically for extracts obtained by SFE, caffeine had to be separated from the oil. The analysis and quantification of diterpenes made by gas chromatography, using himself as cafestol and kahweol external standard, indicated that the oil obtained by SFE showed cafestol 50,180.41 mg/kg oil and kahweol 63,764.32 mg/kg oil in optimal condition. For the PFE maximum concentrations were obtained cafestol 65,876.29 mg/kg oil and kahweol 53,328.47 mg/kg oil and the green coffee oil obtained by the conventional method the concentration of diterpenes was lower than with 7,508 mg cafestol/kg oil kahweol and 12,822 mg/kg oil. The physical measures for the characterization of oil from green coffee beans obtained by SFE and pressing the grains indicated that both the density and the viscosity decrease with increasing temperature. For both oils, the density behavior was similar, to that obtained by the conventional method, the values ranged from 0.94177 g/cm3 (25 °C) to 0.92143 g/cm3 (55 °C) and the oil obtained by the SFE variation was 0.93645 g/cm3 (25 °C) to 0.9157 g/cm3 (55 °C). The dynamic viscosity for the oil obtained by pressing ranged from 127.8798 mPa×s (25 °C) to 35.0510 mPa×s (55 °C) and the oil obtained by SFE, these values were lower, between 84.0411 mPa×s (25 °C) and 24.2555 mPa×s (55 °C). The index of refraction for both oils was similar. In the extraction pressure, (PFE), an optimal value of the extract yield was obtained when methanol was used as solvent at a temperature of 100 °C and static equilibrium time of 20 min. This value was 183% above the average of the results obtained in other tests. At the concentration of these diterpenes extracts, note also that the ethanol was less effective than methanol. In comparing the two innovative processes (SFE and PFE), the concentration of kahweol presents itself as the main component in almost all tests, compared with values of cafestol/kahweol between 0.73 and 0.96. However, the process to be analyzed individually, it is clear that the supercritical fluid extraction indicates that most of the extracts had a concentration higher of kahweol than cafestol.