A Aniba canelilla (H.B.K.) Mez é uma planta aromática proveniente da região amazônica cujo óleo essencial apresenta como componentes majoritários o 1-nitro-2-feniletano e o metileugenol. Apesar das atividades antifúngicas e anti-inflamatórias cientificamente comprovadas e do uso popular do óleo para o tratamento de dermatites, acnes e feridas, não existe até o momento um estudo que verse sobre a quantificação desses compostos na pele. O objetivo deste trabalho foi a validação de um método bioanalítico otimizado por microextração em fase sólida no modo headspace em cromatógrafo gasoso com detector de ionização de chama (HS-SPME-GC-FID) para a determinação do 1-nitro-2-feniletano e metileugenol a partir do óleo essencial em diferentes amostras de estudo de permeação cutânea in vitro. Uma metodologia foi desenvolvida e validada por HS-SPME-GC-FID. A faixa da curva de calibração foi de 2,08 - 207,87 μg.mL-1 para o 1-nitro-2-feniletano e de 0,40-40,41 μg.mL-1 para o metileugenol. A presença de matriz e as características intrínsecas da metodologia de HS-SPME requereram uma transformação da curva de calibração. A transformação logarítmica (Log10) foi então aplicada aos dados e os resultados apresentaram homocedasticidade, resíduos dispersos, coeficiente de determinação (r²> 0,99) e recuperação adequados. Estudos de permeação cutânea foram realizados em células de Franz com diferentes quantidades (20, 100 e 200 μL) de óleo essencial de A. canelilla para avaliar o perfil de permeação e retenção em pele de orelha suína e fluido receptor. A análise das amostras nas condições validadas mostrou uma grande permeação e retenção dos compostos na seguinte ordem: fluido receptor >> derme >> epiderme >> estrato córneo. Verificou-se um aumento progressivo e dependência na retenção com base na quantidade aplicada no compartimento doador e grande retenção principalmente no fluido receptor e derme, resultado este compatível com as características físico-químicas de Log P, afinidade ao ambiente hidrofílico e lipofílico, tamanho e peso molecular do 1-nitro-2-feniletano e metileugenol. Em conclusão, o método proposto por HS-SPME-GC-FID para quantificar os compostos majoritários do óleo essencial de A. canelilla em amostras de pele de orelha suína e fluido receptor foi seletivo, preciso, exato e adequado, e pode ser utilizado em análises futuras. / Aniba canelilla (H.B.K.) Mez is an aromatic plant from the Amazon region whose essential oil has 1-nitro-2-phenylethane and methyleugenol as major compounds. Despite of the scientifically proved antifungal and anti-inflammatory activities and the popular use of oil for the treatment of dermatitis, acnes and wounds, there is no study up to date about the quantification of these compounds in skin samples. The aim of this study was the validation of an optimized bioanalytical method by solid phase microextraction on headspace mode in gas chromatograph with flame ionization detector (HS-SPME-GC-FID) for the determination of 1-nitro-2-phenylethane and methyleugenol from the essential oil in different samples of in vitro cutaneous permeation study. A methodology was developed and validated by HS-SPME-GC-FID. The ranges of calibration curves were 2.08 - 207.87 μg.mL-1 for 1-nitro-2-phenylethane and 0.40 - 40.41 μg.mL-1 for methyleugenol. The presence of matrix and the intrinsic characteristics of the HS-SPME methodology required a transformation to the calibration curves. The logarithmic transformation (Log10) was then applied to the data and the results showed homoscedasticity, dispersed residues, and adequate coefficient of determination (r²> 0.99) and recovery. Skin permeation studies were performed on Franz cells with different amounts (20, 100 and 200 μL) of A. canelilla essential oil to evaluate the skin permeation and retention profile in porcine ear skin and receptor fluid. The analysis of the samples under the validated conditions showed a high permeation and retention of the major compounds in the following order: receptor fluid >> dermis >> epidermis >> stratum corneum. A progressive increase and retention dependence were observed based on the amount applied in the donor compartment, and large retention mainly on the receptor fluid and dermis was observed, in accordance with the physicochemical characteristics of Log P, affinity to the hydrophilic and lipophilic environment, size and molecular weight of 1-nitro-2-phenylethane and methyleugenol. In conclusion, the method proposed by HS-SPME-GC-FID to quantify the major compounds of A. canelilla essential oil in porcine ear skin and receptor fluid samples was selective, accurate, precise and adequate, and can be used in future analyzes.

The two objectives of this work were to develop a simplified method for the analysis of chlorinated organics in water samples and to improve an existing soil remediation technology. The contaminants considered for these studies were chlorinated hydrocarbons because of their relative frequency of appearance at contaminated sites. The first half of this study involved the analysis of chlorinated ethenes by gas chromatography with flame ionization detection (GC-FID). I tested the hypothesis that the FID response factor is the same for all chlorinated ethene compounds. The rationale for this investigation is that if the hypothesis is correct, a single calibration curve can be used for GC/FID analysis of all chlorinated ethene compounds, saving time and money during sample analysis. Based on my measurements, a single calibration curve fits PCE, TCE, and cis-DCE (R2=0.998). However, the apparent slope of the calibration curve for vinyl chloride is approximately 45% lower, indicating that a separate calibration curve must be used to quantify vinyl chloride. I believe this difference in vinyl chloride is due to loss of analyte mass due to volatilization. The second half of the study considered the effect of solvent composition for a soil remediation technology, entitled remedial extraction and catalytic hydrodehalogenation (REACH), developed by Dr. Hun Young Wee and Dr. Jeff Cunningham (Wee and Cunningham, 2008). The objective of this thesis is to convert 1,2,4,5-tetrachlorobenzene (TeCB) to cyclohexane, thus improving on the work of Wee (2007). Recent work by Osborn (2011) tested successfully the use of palladium and rhodium catalysts for this conversion, though it took twelve hours for full conversion. Osborn (2011) performed her experiments in a 50:50 water-ethanol solvent; previous work by Wee and Cunningham (2008) suggests that using a 67:33 water-ethanol composition may dramatically reduce the reaction time. Therefore, the goal of this research was to use palladium and rhodium catalysts with a 67:33 water-ethanol solvent composition, with an aim of reducing the reaction time required to fully convert benzene to cyclohexane. The data suggest that the time required for conversion of the analyte to its product was improved dramatically compared to previous experiments. However, powdered palladium catalyst was used in this study instead of pellet form as in previous studies. The powdered palladium allowed for full conversion of the target chemical, TeCB, to benzene in less than 5 minutes. Benzene was fully converted to cyclohexane within 45 minutes in the batch reactor when a rhodium catalyst was used jointly with palladium. This study suggests that the 67:33 water-ethanol solvent composition be utilized in continuous flow tests in the future to improve the efficiency of the REACH system. The results also suggest that powdered palladium catalyst be considered because of its ability to force the reaction to completion in significantly less time than previous experiments.

A Aniba canelilla (H.B.K.) Mez é uma planta aromática proveniente da região amazônica cujo óleo essencial apresenta como componentes majoritários o 1-nitro-2-feniletano e o metileugenol. Apesar das atividades antifúngicas e anti-inflamatórias cientificamente comprovadas e do uso popular do óleo para o tratamento de dermatites, acnes e feridas, não existe até o momento um estudo que verse sobre a quantificação desses compostos na pele. O objetivo deste trabalho foi a validação de um método bioanalítico otimizado por microextração em fase sólida no modo headspace em cromatógrafo gasoso com detector de ionização de chama (HS-SPME-GC-FID) para a determinação do 1-nitro-2-feniletano e metileugenol a partir do óleo essencial em diferentes amostras de estudo de permeação cutânea in vitro. Uma metodologia foi desenvolvida e validada por HS-SPME-GC-FID. A faixa da curva de calibração foi de 2,08 - 207,87 μg.mL-1 para o 1-nitro-2-feniletano e de 0,40-40,41 μg.mL-1 para o metileugenol. A presença de matriz e as características intrínsecas da metodologia de HS-SPME requereram uma transformação da curva de calibração. A transformação logarítmica (Log10) foi então aplicada aos dados e os resultados apresentaram homocedasticidade, resíduos dispersos, coeficiente de determinação (r²> 0,99) e recuperação adequados. Estudos de permeação cutânea foram realizados em células de Franz com diferentes quantidades (20, 100 e 200 μL) de óleo essencial de A. canelilla para avaliar o perfil de permeação e retenção em pele de orelha suína e fluido receptor. A análise das amostras nas condições validadas mostrou uma grande permeação e retenção dos compostos na seguinte ordem: fluido receptor >> derme >> epiderme >> estrato córneo. Verificou-se um aumento progressivo e dependência na retenção com base na quantidade aplicada no compartimento doador e grande retenção principalmente no fluido receptor e derme, resultado este compatível com as características físico-químicas de Log P, afinidade ao ambiente hidrofílico e lipofílico, tamanho e peso molecular do 1-nitro-2-feniletano e metileugenol. Em conclusão, o método proposto por HS-SPME-GC-FID para quantificar os compostos majoritários do óleo essencial de A. canelilla em amostras de pele de orelha suína e fluido receptor foi seletivo, preciso, exato e adequado, e pode ser utilizado em análises futuras. / Aniba canelilla (H.B.K.) Mez is an aromatic plant from the Amazon region whose essential oil has 1-nitro-2-phenylethane and methyleugenol as major compounds. Despite of the scientifically proved antifungal and anti-inflammatory activities and the popular use of oil for the treatment of dermatitis, acnes and wounds, there is no study up to date about the quantification of these compounds in skin samples. The aim of this study was the validation of an optimized bioanalytical method by solid phase microextraction on headspace mode in gas chromatograph with flame ionization detector (HS-SPME-GC-FID) for the determination of 1-nitro-2-phenylethane and methyleugenol from the essential oil in different samples of in vitro cutaneous permeation study. A methodology was developed and validated by HS-SPME-GC-FID. The ranges of calibration curves were 2.08 - 207.87 μg.mL-1 for 1-nitro-2-phenylethane and 0.40 - 40.41 μg.mL-1 for methyleugenol. The presence of matrix and the intrinsic characteristics of the HS-SPME methodology required a transformation to the calibration curves. The logarithmic transformation (Log10) was then applied to the data and the results showed homoscedasticity, dispersed residues, and adequate coefficient of determination (r²> 0.99) and recovery. Skin permeation studies were performed on Franz cells with different amounts (20, 100 and 200 μL) of A. canelilla essential oil to evaluate the skin permeation and retention profile in porcine ear skin and receptor fluid. The analysis of the samples under the validated conditions showed a high permeation and retention of the major compounds in the following order: receptor fluid >> dermis >> epidermis >> stratum corneum. A progressive increase and retention dependence were observed based on the amount applied in the donor compartment, and large retention mainly on the receptor fluid and dermis was observed, in accordance with the physicochemical characteristics of Log P, affinity to the hydrophilic and lipophilic environment, size and molecular weight of 1-nitro-2-phenylethane and methyleugenol. In conclusion, the method proposed by HS-SPME-GC-FID to quantify the major compounds of A. canelilla essential oil in porcine ear skin and receptor fluid samples was selective, accurate, precise and adequate, and can be used in future analyzes.

A Aniba canelilla (H.B.K.) Mez é uma planta aromática proveniente da região amazônica cujo óleo essencial apresenta como componentes majoritários o 1-nitro-2-feniletano e o metileugenol. Apesar das atividades antifúngicas e anti-inflamatórias cientificamente comprovadas e do uso popular do óleo para o tratamento de dermatites, acnes e feridas, não existe até o momento um estudo que verse sobre a quantificação desses compostos na pele. O objetivo deste trabalho foi a validação de um método bioanalítico otimizado por microextração em fase sólida no modo headspace em cromatógrafo gasoso com detector de ionização de chama (HS-SPME-GC-FID) para a determinação do 1-nitro-2-feniletano e metileugenol a partir do óleo essencial em diferentes amostras de estudo de permeação cutânea in vitro. Uma metodologia foi desenvolvida e validada por HS-SPME-GC-FID. A faixa da curva de calibração foi de 2,08 - 207,87 μg.mL-1 para o 1-nitro-2-feniletano e de 0,40-40,41 μg.mL-1 para o metileugenol. A presença de matriz e as características intrínsecas da metodologia de HS-SPME requereram uma transformação da curva de calibração. A transformação logarítmica (Log10) foi então aplicada aos dados e os resultados apresentaram homocedasticidade, resíduos dispersos, coeficiente de determinação (r²> 0,99) e recuperação adequados. Estudos de permeação cutânea foram realizados em células de Franz com diferentes quantidades (20, 100 e 200 μL) de óleo essencial de A. canelilla para avaliar o perfil de permeação e retenção em pele de orelha suína e fluido receptor. A análise das amostras nas condições validadas mostrou uma grande permeação e retenção dos compostos na seguinte ordem: fluido receptor >> derme >> epiderme >> estrato córneo. Verificou-se um aumento progressivo e dependência na retenção com base na quantidade aplicada no compartimento doador e grande retenção principalmente no fluido receptor e derme, resultado este compatível com as características físico-químicas de Log P, afinidade ao ambiente hidrofílico e lipofílico, tamanho e peso molecular do 1-nitro-2-feniletano e metileugenol. Em conclusão, o método proposto por HS-SPME-GC-FID para quantificar os compostos majoritários do óleo essencial de A. canelilla em amostras de pele de orelha suína e fluido receptor foi seletivo, preciso, exato e adequado, e pode ser utilizado em análises futuras. / Aniba canelilla (H.B.K.) Mez is an aromatic plant from the Amazon region whose essential oil has 1-nitro-2-phenylethane and methyleugenol as major compounds. Despite of the scientifically proved antifungal and anti-inflammatory activities and the popular use of oil for the treatment of dermatitis, acnes and wounds, there is no study up to date about the quantification of these compounds in skin samples. The aim of this study was the validation of an optimized bioanalytical method by solid phase microextraction on headspace mode in gas chromatograph with flame ionization detector (HS-SPME-GC-FID) for the determination of 1-nitro-2-phenylethane and methyleugenol from the essential oil in different samples of in vitro cutaneous permeation study. A methodology was developed and validated by HS-SPME-GC-FID. The ranges of calibration curves were 2.08 - 207.87 μg.mL-1 for 1-nitro-2-phenylethane and 0.40 - 40.41 μg.mL-1 for methyleugenol. The presence of matrix and the intrinsic characteristics of the HS-SPME methodology required a transformation to the calibration curves. The logarithmic transformation (Log10) was then applied to the data and the results showed homoscedasticity, dispersed residues, and adequate coefficient of determination (r²> 0.99) and recovery. Skin permeation studies were performed on Franz cells with different amounts (20, 100 and 200 μL) of A. canelilla essential oil to evaluate the skin permeation and retention profile in porcine ear skin and receptor fluid. The analysis of the samples under the validated conditions showed a high permeation and retention of the major compounds in the following order: receptor fluid >> dermis >> epidermis >> stratum corneum. A progressive increase and retention dependence were observed based on the amount applied in the donor compartment, and large retention mainly on the receptor fluid and dermis was observed, in accordance with the physicochemical characteristics of Log P, affinity to the hydrophilic and lipophilic environment, size and molecular weight of 1-nitro-2-phenylethane and methyleugenol. In conclusion, the method proposed by HS-SPME-GC-FID to quantify the major compounds of A. canelilla essential oil in porcine ear skin and receptor fluid samples was selective, accurate, precise and adequate, and can be used in future analyzes.

Seminario de Título entregado a la Universidad de Chile en cumplimiento parcial de los requisitos para optar al Título de Química Ambiental. / Los suelos se han constituido como el principal sumidero de metales pesados y otros contaminantes producto de causas naturales y actividades antropogénicas. Ejemplo de estas actividades son las mineras que liberan metales pesados, tales como Hg+2, Pb+2, Cu+2 y Zn+2, y las plantas petroquímicas que producen hidrocarburos aromáticos como Benceno, Tolueno, Etilbenceno, y Xilenos (BTEX) a partir de la fracción volátil del petróleo. El objetivo de este seminario de título consiste en validar un método analítico para la cuantificación de benceno en microcosmos conformados por muestras de suelo co-contaminado contenidas en viales, a los cuales se le adiciona una bacteria especializada. De esta forma, se desarrolla una metodología para determinar benceno remanente en suelo, carente de solventes orgánicos, simple y costo-efectiva, mediante un sistema de extracción de espacio de cabeza acoplado a un cromatógrafo de gases junto a un detector de ionización de llama (HS-GC-FID), que permita monitorear la cinética de remoción de benceno en un suelo co-contaminado con Hg (II), durante un proceso de biorremediación bacteriana utilizando la cepa modificada genéticamente Cupriavidus metallidurans MSR33 que es altamente resistente a mercurio. De esta manera, en el presente trabajo se utilizó un suelo contaminado con benceno y mercurio como una aproximación a una situación real de co-contaminación, donde se aplicó una técnica de remediación a través de la utilización de bacterias especializadas capaces de remover benceno en presencia de mercurio. La medición de benceno con la metodología validada permitió monitorear la cinética de biorremediación utilizando la bacteria C. metallidurans MSR33 entregando información rápida y veraz al aplicar directamente en viales con microcosmos de suelo contaminados con Benceno 200 mg×Kg-1 y Hg (II) 2 mg×Kg-1. La determinación de benceno permitió demostrar que este proceso de biorremediación conforma una novedosa tecnología costo-efectiva y amigable con el medioambiente, aplicable a suelos impactados con BTEX en presencia de metales tóxicos. / Soils have been established as the main sink for heavy metals and other pollutants due to natural causes and anthropogenic activities. Examples of these are mining activities which release heavy metals, such as Hg+2, Pb+2, Cu+2 and Zn+2, and petrochemical plants which produce aromatic hydrocarbons such as Benzene, Toluene, Ethylbenzene, and Xylene (BTEX) from volatile fractions of petroleum. The aim of this seminar consists of validate an analytical method for the quantification of benzene in microcosms consisting on vials containing co-contaminated soil samples, which are inoculated with a specially adapted bacterial culture. The methodology was developed in order to measure benzene concentrations remaining in soil in an organic solvent-free, simple and cost-effective manner. This was carried by means of a headspace extraction system coupled to a gas chromatograph with flame ionization detector (HS-GC-FID), that allowed to monitor the kinetics of benzene removal in a soil co-contaminated with Hg (II), during the process of bacterial bioremediation with the genetically modified strain Cupriavidus metallidurans MSR33 that is highly resistant to mercury. Thus, soil samples polluted with benzene and mercury were used on this research as a practical approach to a real co-contamination scenario. A remediation technique was applied by means of bacteria specially adapted to remove benzene from soil in presence of mercury. The measurement of benzene with the validated methodology allowed to monitor the kinetics of bioremediation using the C. metallidurans MSR33 bacterial strain delivering fast and accurate information when applied directly in vials with soil microcosms contaminated with Benzene 200 mg×Kg-1 and Hg (II) 2 mg×Kg-1. The determination of benzene allowed to demonstrate that this process of bioremediation forms a novel, cost-effective and environmental-friendly technology, potentially useful for treatment of soils impacted with BTEX in the presence of toxic metals.

The aim of this diploma thesis was to determine the effect of the addition of several types of local wines in the production of wine sausages. Selected wine varieties were characterized using basic and modern analytical methods (liquid chromatography with a diode array detector) and the obtained results were compared with the results of sensory analysis.The sensory analysis was divided into two parts, the first evaluated six non-traditional wine varieties used in the production of wine sausages and the second part evaluated samples with the addition of the traditionally used wine variety Veltlínské zelené, but in three different volumes. Using the Statistica program, the data obtained from the sensory analysis were processed and the mutual correlation was evaluated. Using the gas chromatography method with a flame ionization detector the fatty acid content of a standard sample of wine sausage has been determined. Based on all the determined results, the best wine variety for the production of wine sausages was chosen, it was the blue grape variety André.

This master´s thesis deals with production and characterization of fresh cheese with addition of herb extracts, specifically extracts from Plectranthus amboinicus and Echinacea purpurea. The purpose of this thesis was to prepare samples with higher sensory quality and nutritional value. Produced cheeses were characterized in terms of volatile (aroma) substances, fatty acid content, total phenolic substances, antioxidant activity and sensory quality. Cheese sample with added extract from E. purpurea reached the highest value of total phenolic compounds (0,172 ± 0,019 mgGAEg-1 of cheese) and antioxidant activity (91,2 ± 2,2 gTEACg-1 of cheese). Cheese sample with added extract from P. amboinicus, compared to standard cheese sample (without added extract) also reached higher values of total phenolic compounds and antioxidant activity. 43 volatile (aroma) compounds were identified in both samples with added extracts (P. amboinicus, E. purpurea), in standard cheese sample it was 33. In sample with E. purpurea camphor was present in high amount (15,23 %) responsible for the camphor aroma, camphor was also identified in small amount in sample with P. amboinicus. Highest volatile matter content was identified in sample with P. amboinicus. Standard cheese sample was rated as the best from the sensory perspective. Addition of extracts had negative effect on color, taste, and aroma perception of cheese. Strongest herbal taste was identified in sample with E. purpurea, brownish color of this sample manifested in lower pleasantness of color. Obtained results show, that extract addition increases the nutritional value of cheese (higher content of aromatic substances and phenolic compounds, higher antioxidant activity), but lowers sensory quality and overall consumer acceptability.

This thesis deals with the production and characterization of cheese analogues, i.e. products where some of the dairy components are partially or completely replaced by a non-dairy component. Model samples – Eidam type cheese and analogue with the addition of pistachio oil, were produced by a conventional technological process at MENDELU in Brno. Free and bound fatty acids, volatile substances and sensory quality were selected and monitored as the main characterizing parameters of the sample quality. The aim was to characterize the differences between the samples caused by the addition of nut oil in the monitored parameters. The HS-SPME-GC-MS method was used for the identification and semi-quantification of volatiles. The extraction of fats from the samples was performed with a mixture of solvents (diethyl ether, petroleum ether), fatty acids were converted to methyl esters by acid esterification with methanolic solution of boron trifluoride as a catalyst and subsequently identified and quantified by GC-FID. Descriptive scales and comparisons with standard were used to sensory evaluate the appearance, color, texture, odor, taste, and overall acceptability of the samples. The structure was monitored by electron microscopy. The results show that the addition of pistachio oil affects the sensory properties, the maturation process (higher content of volatile substances), the nutritional value (increased content of unsaturated fatty acids) and the stability of the product.

This thesis is focused on potentially allergenic substances (PASs) in cosmetics. Their specific features, methods of control and observation are described. There are also mentioned dermatology problems connected with usage of perfumed cosmetic products in which the allergenic ingredients can be found and possibilities of their regulation. Fragrances are mixtures of natural or synthetic character which give the aroma. In nature, fragrances are produced mainly by plants. More than one hundred fragrances have been identified as allergens. Twenty six of them were declared as PASs. Allergenic components of perfume compositions are the main cause of allergic contact dermatitis. Experimental part of the thesis is focused on the procedure of analysis of specific allergenic fragrances (?-Amylcinnamaldehyde, Anisealcohol, Benzylsalicylate, Cinnamaldehyde, ?-Hexylcinnamaldehyde, 7-Hydroxycitronellal and Methyl-2-octynoate). Headspace-Solid Phase Microextraction connected with Gas Chromatography-Flame Ionisation Detector (HS-SPME-GC-FID) was used for extraction, analysis and identification of these substances. The method was validated and applied on the spectrum of real samples of cosmetic products.

This master thesis deals with the assessment of aroma active compounds in fruits of currant (Ribes spp.). The theoretical part was focused on basic characteristics and chemical composition of currants, as well as the basic principles of techniques used: solid phase micro extraction (SPME), gas chromatography (GC) and sensory analysis. The main aim of the experimental part was to determine aroma active compounds in currants using SPME-GC-FID method. The profile and scale tests were used for assessment of sensory quality and tastiness of fruits. In total 18 varieties, among them 6 white, 6 red and 6 black currants, were evaluated. Fifty aroma active compounds were identified including 21 alcohols, 10 aldehydes, 2 acids, 8 ketones and 9 esters. The results were statistically evaluated using principal component analysis (PCA) to investigate the influence of single compounds on sensory quality of fruits.