Valorisation of vegetable oil deodorizer distillate by enzymatic reaction and membrane processing

Dissertação para obtenção do Grau de Doutora em Engenharia Química e Bioquímica

Production of polyhydroxyalkanoates from oil-containing substrates

Different oil-containing substrates, namely, used cooking oil (UCO), fatty acids-byproduct from biodiesel production (FAB) and olive oil deodorizer distillate (OODD) were tested as inexpensive carbon sources for the production of polyhydroxyalkanoates (PHA) using twelve bacterial strains, in batch experiments. The OODD and FAB were exploited for the first time as alternative substrates for PHA production. Among the tested bacterial strains, Cupriavidus necator and Pseudomonas resinovorans exhibited the most promising results, producing poly-3-hydroxybutyrate, P(3HB), form UCO and OODD and mcl-PHA mainly composed of 3-hydroxyoctanoate (3HO) and 3-hydroxydecanoate (3HD) monomers from OODD, respectively. Afterwards, these bacterial strains were cultivated in bioreactor. C. necator were cultivated in bioreactor using UCO as carbon source. Different feeding strategies were tested for the bioreactor cultivation of C. necator, namely, batch, exponential feeding and DO-stat mode. The highest overall PHA productivity (12.6±0.78 g L-1 day-1) was obtained using DO-stat mode. Apparently, the different feeding regimes had no impact on polymer thermal properties. However, differences in polymer‟s molecular mass distribution were observed. C. necator was also tested in batch and fed-batch modes using a different type of oil-containing substrate, extracted from spent coffee grounds (SCG) by super critical carbon dioxide (sc-CO2). Under fed-batch mode (DO-stat), the overall PHA productivity were 4.7 g L-1 day-1 with a storage yield of 0.77 g g-1. Results showed that SCG can be a bioresource for production of PHA with interesting properties. Furthermore, P. resinovorans was cultivated using OODD as substrate in bioreactor under fed-batch mode (pulse feeding regime). The polymer was highly amorphous, as shown by its low crystallinity of 6±0.2%, with low melting and glass transition temperatures of 36±1.2 and -16±0.8 ºC, respectively. Due to its sticky behavior at room temperature, adhesiveness and mechanical properties were also studied. Its shear bond strength for wood (67±9.4 kPa) and glass (65±7.3 kPa) suggests it may be used for the development of biobased glues. Bioreactor operation and monitoring with oil-containing substrates is very challenging, since this substrate is water immiscible. Thus, near-infrared spectroscopy (NIR) was implemented for online monitoring of the C. necator cultivation with UCO, using a transflectance probe. Partial least squares (PLS) regression was applied to relate NIR spectra with biomass, UCO and PHA concentrations in the broth. The NIR predictions were compared with values obtained by offline reference methods. Prediction errors to these parameters were 1.18 g L-1, 2.37 g L-1 and 1.58 g L-1 for biomass, UCO and PHA, respectively, which indicates the suitability of the NIR spectroscopy method for online monitoring and as a method to assist bioreactor control. UCO and OODD are low cost substrates with potential to be used in PHA batch and fed-batch production. The use of NIR in this bioprocess also opened an opportunity for optimization and control of PHA production process.

Estudo do comportamento da lipase comercial Lipozyme RM IM em reações de esterificação para obtenção de biodiesel

The aim of this work was to study monoalkyl ester synthesis catalyzed by immobilized lipase Lipozyme RM IM via the esterification reaction. Yields of over 90% were obtained with butanol in esterification reactions with oleic acid. In the reactions with deodorizer distillates of vegetable oils and butanol, the conversion obtained was greater than 80% after 2.5 h. For the esterification reaction of palm fatty acid deodorizer distillate (PFAD) and butanol, seven reuse cycles of Lipozyme RM IM were carried out and the final conversion was 42% lower than the initial conversion.

Influence of palm fatty acid distillate on rumen degradability and protozoa population in buffaloes

Oil and fat as energy sources at low cost are relevant in ruminant nutrition. The aim of this study was to evaluate the effects of palm fatty acid distillate (PFAD) on the degradability and ciliate protozoa population in buffalo. Four rumen fistulated buffaloes were fed a basal diet in a Latin square (4x4) design trial. Treatments were designed with four of different levels of PFAD added directly into the rumen: 0; 200; 420 and 500 g/animal/d. High levels of PFAD (420 and 500 g/d) promoted higher degradation of the soluble fraction and lower in potentially degradable fraction of dry matter (DM) and neutral detergent fibre (NDF) with lower values of potential and effective degradability in two evaluated grasses, bermudagrass and brachiariagrass. Significant decreases in the total number of protozoa/mL of rumen content, Entodinium and ciliates belonging to subfamily Diplodiniinae were observed at higher level of PFDA addition in the rumen. Also, Epidinium and Holotrich ciliates disappeared from the rumen. Significant correlations were observed of the ciliate concentration and composition as a function of dietary lipids content. Entodinium composition increased from 68.0% to 99.6% and Diplodiniinae reduced from 30.4% to 0.4% with increasing PFAD level indicating higher fat toxicity effect on the Diplodiniinae ciliates than Entodinium species and direct action of the larger ciliates on the fibre degradation.

Effect of replacing calcium salts of palm oil distillate with rapeseed oil, milled or whole rapeseeds on milk fatty-acid composition in cows fed maize silage-based diets

Inclusion of rapeseed feeds in dairy cow diets has the potential to reduce milk fat saturated fatty acid (SFA) and increase cis-monounsaturated fatty acid (cis-MUFA) content but effectiveness may depend on the form in which the rapeseed is presented. Four mid-lactation Holstein dairy cows were allocated to four maize silage-based dietary treatments according to a 4 x 4 Latin Square design, with 28-day experimental periods. Treatments consisted of a control diet (C containing 49 g/kg dry matter (DM) of calcium salts of palm oil distillate (CPO), or 49 g/kg DM of oil supplied as whole rapeseeds (WR), rapeseeds milled with wheat (MR) or rapeseed oil (RO). Replacing CPO with rapeseed feeds had no effect (P > 0.05) on milk fat and protein content, while milk yields were higher (P < 0.05) for RO and MR compared with WR (37.1, 38.1 and 34.3 kg/day, respectively). Substituting CPO with RO or MR reduced (P < 0.05) milk fat total SFA content (69.6, 55.6, 71.7 and 61.5 g/100g fatty acids for C, RO, WR and MR, respectively) and enhanced (P < 0.05) milk cis-9 18:1 MUFA concentrations (corresponding values 18.6, 24.3, 17.0 and 23.0 g/100g fatty acids) compared with C and WR. Treatments RO and MR also increased (P < 0.05) milk trans-MUFA content (4.4, 6.8, 10.5 g/100g fatty acids, C MR and RO, respectively). A lack of significant changes in milk fat composition when replacing CPO with WR suggests limited bioavailability of fatty acids in intact rapeseeds. In conclusion, replacing a commercial palm oil-based fat supplement in the diet with milled rapeseeds or rapeseed oil represented an effective strategy to alter milk fatty acid composition with the potential to improve human health. Inclusion of processed rapeseeds offered a good compromise for reducing milk SFA and increasing cis-MUFA, whilst minimising milk trans-MUFA and negative effects on animal performance.

Effect of replacing calcium salts of palm oil distillate with extruded linseeds on milk fatty acid composition in Jersey and Holstein cows

Clinical and biomedical studies have provided evidence for the critical role of n-3 fatty acids on the reduction of chronic disease risk in humans, including cardiovascular disease. In the current experiment, the potential to enhance milk n-3 content in two breeds with inherent genetic differences in mammary lipogenesis and de novo fatty acid synthesis was examined using extruded linseeds. Six lactating cows (three Holstein and three Jersey) were used in a two-treatment switchback design with 3 × 21-day experimental periods to evaluate the effect of iso-energetic replacement of calcium salts of palm oil distillate (CPO) in the diet (34 g/kg dry matter (DM)) with 100 g/kg DM extruded linseeds (LIN). For both breeds, replacing CPO with LIN had no effect (P > 0.05) on DM intake or milk yield, but reduced (P < 0.05) milk fat and protein yield (on average, from 760 to 706 and 573 to 552 g/day, respectively). Relative to CPO, the LIN treatment reduced (P < 0.01) total saturated fatty acid content and enhanced (P < 0.001) 18:3n-3 in milk, whereas breed by diet interactions were significant for milk fat 16:0, total trans fatty acid and conjugated linoleic acid concentrations. Increases in 18:3n-3 intake derived from LIN in the diet were transferred into milk with a mean marginal transfer efficiency of 1.8%. Proportionate changes in milk fatty acid composition were greater in the Jersey, highlighting the importance of diet–genotype interactions on mammary lipogenesis. More extensive studies are required to determine the role of genotype on milk fat composition responses to oilseeds in the diet.

Effects of nutrition on digestión efficiency and gaseous emissions from slurry in growing pigs: III. Influence of varying the dietary level of calcium soap of palm fatty acids distillate with or without orange pulp supplemention

The aim of this study was to establish the relationships between faecal fat concentration and gaseous emissions from pig slurry. Five diets were designed to meet essential nutrient requirements: a control and four experimental feeds including two levels (35 or 70 g/kg) of calcium soap fatty acids distillate (CSP) and 0 or 200 g/kg of orange pulp (OP) combined in a 2 × 2 factorial structure. Thirty growing pigs (six per treatment) were used to measure dry matter (DM) and N balance, coefficients of total tract apparent digestibility (CTTAD) of nutrients, faecal and urine composition and potential emissions of ammonia (NH3) and methane (CH4). Increasing dietary CSP level decreased DM, ether extract (EE) and crude protein (CP) CTTAD (by 4.0, 11.1 and 3.5%, respectively, P < 0.05), but did not influence those of fibrous constituents. It also led to a decrease (from 475 to 412 g/kg DM, P < 0.001) of faecal concentration of neutral detergent fibre (aNDFom) and to an increment (from 138 to 204 g/kg, P < 0.001) of EE in faecal DM that was related to greater CH4 emissions, both per gram of organic matter (P = 0.021) or on a daily basis (P < 0.001). Level of CSP did not affect N content in faeces or urine, but increased daily DM (P < 0.001), and N (P = 0.031) faecal excretion with no effect on urine N excretion. This resulted in lesser (P = 0.036) NH3 potential emission per kg of slurry. Addition of OP decreased CTTAD of EE (by 7.9%, P = 0.044), but increased (P < 0.05) that of all the fibrous fractions. As a consequence, faecal EE content increased (from 165 to 177 g/kg DM; P = 0.012), and aNDFom decreased greatly (from 483 to 404 g/kg DM, P < 0.001), which in all resulted in a lack of effect of OP on CH4 potential emission. Inclusion of OP in the diet also led to a significant decrease of CP CTTAD (by 6.85%, P < 0.001), and to an increase of faecal CP concentration (from 174 to 226 g/kg DM, P < 0.001), with no significant influence on urine N content. These effects resulted in higher N faecal losses, especially those of the undigested dietary origin, without significant effects on potential NH3 emission. No significant interactions between CSP and OP supplementation were observed for the gaseous emissions measured.

Batch distillation: Better at constant or variable reflux?

Due to its outstanding flexibility, batch distillation is still widely used in many separation processes. In the present work, a comparison between constant and variable reflux operations is studied. Firstly, a mathematical model is developed and then validated through comparison between predicted and experimental results accomplished in a lab-scale apparatus. Therefore, case studies are performed through mathematical simulations. It is noted that the most economical form of batch distillation is at constant overhead product composition, keeping the flow rate of vapor from the top of the column constant. (C) 2010 Elsevier B.V. All rights reserved.

Comparison of the chemical composition of the essential oil and the water soluble oil of Baccharis dracunculifolia DC. (Asteraceae)

This study aimed to evaluate the chemical composition of Baccharis dracunculifolia essential oil and the water soluble oil obtained by steam distillation that were analyzed by GC and GUMS. in the first hour of distillation, B. dracunculifolia aerial parts yielded 0.08% oil and in the second hour, 0.27%. The oil recovered from the distillate water yielded 0.18 g/L in the first hour and 0.44 g/L in the second hour of distillation. The main volatile compounds identified in the distillate water were aromatic compounds and sesquiterpene alcohols.

Avaliação da influência de antioxidantes na estabilização de biodiesel

O biodiesel é uma fonte de energia renovável, que se pode obter pela transformação dos resíduos domésticos, e é ambientalmente inócuo, e fácil de transportar, pois tem um ponto de fulgor elevado. Atualmente tem-se focado a atenção nos efeitos da oxidação do biodiesel causados pelo contato com o ar ambiente durante o seu armazenamento. Os produtores, fornecedores e consumidores, pretendem garantir que a qualidade do biodiesel e das suas misturas com combustíveis destilados do petróleo, mantém-se durante longos períodos de armazenamento. A maioria dos óleos vegetais e das gorduras animais, usados como matéria – prima, são triacilgliceróis com grupos ácido gordos de cadeia longa (C16 – C18) ligados por ligações éster a uma estrutura de glicerol. O objetivo deste estudo foi avaliar a eficiência do ácido gálhico e seus derivados alquilo ésteres aumentando a resistência relativa à oxidação do ácido linoléico, inibindo a peroxidação ácido gordos de cadeia longa insaturados. Outro objetivo foi estudar a taxa de inibição da auto-oxidação do ácido linoleico. Foi, ainda estudado o efeito que a concentração do antioxidante, tinha na estabilidade do ácido linoleico. No caso do ácido gálhico verificou-se que o aumento para o dobro da concentração do antioxidante utilizada, obtinha-se quase o dobro da taxa de inibição da oxidação do ácido linoleico. A auto-oxidação de ácido linoleico é acompanhada pela formação do seu dieno conjugado, o qual foi medido sua absorvância, durante 7 dias, por espetrofotometria de absorção UV a 234 nm. Uma diminuição da taxa de formação de dieno conjugado, indica o aumento da atividade antioxidante do composto adicionado à micela de ácido linoleico. Os resultados obtidos permitem concluir que de todos os antioxidantes testados o galhato de butilo é o que possibilita uma maior inibição da oxidação do ácido linoleico para as duas concentrações de antioxidantes testadas (0,1 mM e 1mM), obteve-se uma percentagem de inibição do ácido linoleico de 54,0% e 63,6%, respetivamente. O estudo comparativo da estabilização do ácido linoleico com o antioxidante de referência, o butil -hidroxitolueno, mostrou que este composto tem um poder de estabilização inferior a qualquer dos antioxidantes estudados. Os resultados deste estudo demonstraram que a utilização de compostos fenólicos, em especial o galhato de butilo, constitui uma boa alternativa para a estabilização de matrizes lipídicas, nomeadamente de combustíveis como o biodiesel.

Maximização da produção de paraxileno

Pretendeu-se, com o presente trabalho, efectuar um estudo sobre a possibilidade de aumentar a produção de paraxileno na unidade processual existente na refinaria da Galp no Porto. Foram estudados o conjunto de colunas T-0306 e T-0307 e a coluna T-0303, recorrendo ao uso do AspenPlus, na realização da simulação do processo nas condições actuais e para a realização da análise de sensibilidade, que permitiu verificar se o equipamento existente possui capacidade para satisfazer as necessidades exigidas pelo processo quando se pretende aumentar a produção. Com a simulação do processo, foi verificado que este já se encontra a laborar em condições óptimas. Na realização da análise de sensibilidade foram variados para o conjunto T-0306 e T- 0307, o caudal de alimentação entre 62780 e 95000 kg/h, o caudal de destilado da T-0306 entre 15833 e 23911 kg/h e o caudal de destilado da T-0307 entre 139 e os 235 kg/h. No caso da T-0303, foi variado o caudal de refinado entre 201240 e 304600 kg/h, o caudal de destilado e o caudal da corrente lateral, variam entre 77290 a 116930 kg/h. Pretendeu-se obter soluções que conseguissem cumprir as especificações relativas aos produtos obtidos, nomeadamente paraxileno com uma pureza de 99,6% e uma recuperação de 99%, tolueno com uma pureza de 97% e uma recuperação de 90% (T-0306 e T-0307). No caso da T- 0303, obter paraxileno com uma pureza de 2,23% e paradietilbenzeno com uma pureza de 99,0% e ambos com uma recuperação de 99%. Para as simulações que cumpriram as especificações anteriores, verificou-se se a energia transferida em aeroarrefecedores, fornalhas e permutadores excedem em 25% o seu valor de projecto. Em caso afirmativo isso significou o acréscimo de uma peça de equipamento idêntica à existente. A análise de sensibilidade permitiu concluir a separação é possível para os caudais actuais de extracto e refinado e para um aumento destes de 32, 40 e 50%. Foi ainda possível concluir que para os caudais actuais é necessária a colocação de novos aeroarrefecedores para as colunas T-0306 e T-0303 e de uma nova fornalha para a coluna T-0303. Para um aumento dos caudais de 32% para além das alterações referidas anteriormente é necessária a colocação de um novo aeroarrefecedor na coluna T-0307 e de uma nova fornalha da coluna T-0306. Se aumentarmos os caudais em 40% será também necessário o acréscimo de permutadores para a coluna T-0307. Para o aumento de 50%, os calores de permuta são excedidos em bastante mais do que 25%, em todos os aeroarrefecedores, permutadores e fornalhas. Uma vez que todas as colunas se encontram a funcionar acima da sua capacidade, concluiu-se que em ambos os casos seria necessária a montagem de uma segunda linha em paralelo e igual à existente. Implicando com esta alteração um investimento total de 10.439.574,36€.

Determination of 24 pesticide residues in fortified wines by solid-phase microextraction and gas chromatography tandem mass spectrometry

The present work describes a solid-phase microextraction (SPME) gas chromatography_tandem mass spectrometry (MS/MS) method to quantify 24 pesticides in fortified white wine and fortified red wine. In this study “fortified wine” refers to a wine in which fermentation is arrested before completion by alcohol distillate addition, allowing sugar and alcoholic contents to be higher (around 80-100 g/L total sugars and 19-22% alcohol strength (v/v)). The analytical method showed good linearity, presenting correlation coefficients (R2) ≥ 0.989 for all compounds. Limits of detection (LOD) and quantitation (LOQ) in the ranges of 0.05-72.35 and 0.16-219.23 μg/L, respectively, were obtained. LOQs are below the maximum residue levels (MRL) set by European Regulation for grapes. The proposed method was applied to 17 commercial fortified wines. The analyzed pesticides were not detected in the wines tested.

Traditional Chinese materia medica: a retrospect and prospect

For thousands of years, traditional medicine and remedies have been practed and used in the fight against disease in China. They have proved to be valuable and the distillate of vast historical experience based on field-tested human experiments, long-term observations and clinical trials. The Chinese people belive that traditional medicine is consistent with theirown culture. Endowed with a unique theoretical system and provided outstanding clinical results, traditional Chinese medicine continues to play an important role in helping the Chinese nation flourish. The recent study of traditional medicinal plants in Chine has given us confidence that what was recorded in ancient medical literature through empirical observations is indeed still coindicent with the concepts of modern chemistry, pharmacology and medicine. The task of revealing what is valid and efficacious should be retained, and what is mythic and invalid should be discarded in traditional Chinese medicine may require scientific research lasting for several generations. Therfore, multidisciplinary cooperation and international collaboration in this field would be essential. Systematic coordination of work in traditional medicine by word organizations, national governments, private foundations and individual scientists is a requisite as well.

Prophylaxis with resin in wood ants

Animals may use plant compounds to defend themselves against parasites. Wood ants, Formica paralugubris, incorporate pieces of solidified conifer resin into their nests. This behaviour inhibits the growth of bacteria and fungi in nest material and protects the ants against some detrimental microorganisms. Here, we studied the resin-collecting behaviour of ants under field and laboratory conditions. First, we focused on an important assumption of the self-medication hypothesis, which is that the animals deliberately choose the active plant material. In field cafeteria tests, the ants indeed showed a strong preference for resin over twigs and stones, which are building materials commonly encountered in their environment. We detected seasonal variation in the choice of ants: the preference for resin over twigs was more pronounced in spring than in summer, whereas in autumn the ants collected twigs and resin at equal rates. Second, we found almost similar seasonal patterns when comparing the collecting rates of pieces of wood that had been impregnated with turpentine (a distillate of oleoresin) and untreated pieces of wood, which reveals that the preference for resin is based on odour cues. Third, we tested whether the collection of resin is prophylactic or therapeutic. We found that the relative collection rate of resin versus stones did not depend on an experimental infection with the entomopathogenic fungus Metarhizium anisopliae in laboratory colonies. Together, these results show that the ants deliberately choose the resin and suggest that resin collection is prophylactic rather than therapeutic.