Enzymatic transformations in organic solvents have received increasing attention over the past 10 years and lipases have become by far the most popular enzymes in this area.The initial aim of the research was to assess the effect of small modifications to the acyl donor in the transesterification reactions mediated by the Candida cylindracea lipase. 2.2.2-Trichloroethyl butyrate (TCEB) was used as a standard for the rate studies. In the event the acyl donor, trichloroethyl methoxyacetate (TCEMA), accelerated the transesterification reaction with hexan-1-ol by an order of magnitude over that with TCEB. This observation, and the absence of an acceleration with trichloroethyl methoxypropionate (and ethyl 2- fluorobutyrate over ethyl butyrate) suggested that the effect is due to the ~oxygen. A solvent activity profile indicated that the most hydrophobic solvents supported faster initial rates. This was attributed to the ability of the hydrophilic solvents to strip the hydrated water from the enzyme surface thus deactivating it. The switch to organic solvents allowed a wider temperature range to be studied. For the reactions between heptan-2-ol and TCEMA the reaction could be conducted in the temperature range -23 C to 80 C. It was of interest to assess how the alcohol moiety effected the transesterification reaction. A series of alcohols were presented to the enzyme and a pattern emerged with substrates containing an acetylene functionality being processed faster than those with a vinyl group, which were faster than those containing a methyl group (all other groups being the same). A series of heterocyclic alcohols were presented to the enzyme and it was observed that the order of reaction was thiophene > furan > pyridyl. The secondary alcohols in this series, 2-thiopheneethan-1-ol and o pyridylethan-1-ol, were resolved at various temperatures from -1soc to sooc with no variation in. enantioselectivity. These are the first resolutions to be accomplished at temperatures below zero degrees.

<p>Biofuels are a developing kind of fuel whose origin is biomass. Among them, many different kind of fuels can be found: bioethanol, biobutanol, biodiesel, vegetable oils, biomethanol, pyrolysis oils, biogas, and biohydrogen. This thesis work is focused on the production of biodiesel, which can be used in diesel engines as a substitute for mineral diesel. Biodiesel is obtained from different kinds of oils, both from vegetable and animal sources. However, vegetable oils are preferred because they tend to be liquid at room temperature.</p><p>The process to obtain biodiesel implies first a reaction between the oil and an alcohol, using a catalyst and then a sedimentation, where the biodiesel and the glycerol, the two products that are obtained, can be separated because of their difference in density. After the separation, raw biodiesel is obtained and a treatment with either water bubbling or dry cleaning products is needed to obtain the product which will be ready to use.</p><p>Many methods are available for the production of biodiesel, most of them require heat for the transesterification reaction, which converts the oil into biodiesel. Apart from that, in many cases biodiesel is produced by big companies or by individuals but using complicated and expensive installations.</p><p>This work is an attempt to develop a way of producing biodiesel without any use of external heat, using a simple procedure which could be used by people with a low knowledge of chemistry or chemical processes. It also seeks to set an example on how biodiesel can be easily made by oneself without the use of any industrial systems, with a low budget and limited need of supervision over the process.</p><p>In order to achieve that, many trials were undertaken, introducing changes in the different parameters that are responsible for the changes in the final product. Among them, changes in the amount and type of catalyst, the way the catalyst is added, the type of oil used, the time of reaction and the temperature were made. Apart from that, different types of cleaning were tried, starting by water cleaning and then using powder type products, Magnesol, D-Sol and Aerogel. A centrifuge was also tried to test its utility when separating impurities from liquids or different liquid phases. The results of the different trials were analysed using various tests, the most important being the 3:27 test, the solubility test, the soap titration and pH measurements.</p><p>To sum up, it could be said that the investigation was a success, since it was proved that biodiesel can be made without the use of any external heat with both alkali and acid catalysts, as well as with different ways of adding the catalyst. As for the cleaning, good results were obtained with both dry products and water cleaning, since the soap content of the biodiesel was reduced in both cases. Apart from that, the centrifuge proved to be valid to eliminate impurities from raw oil.</p>

Biodiesel is a renewable, biodegradable, environmentally benign fuel for use in the diesel engines. It can be produced from renewable sources such as vegetable oils or animal fats. Although this fuel has gained worldwide recognition for many years, it is not being widely commercialized mainly because it is more expensive than petroleum diesel. A cheaper feedstock, such as fryer grease, may be used to improve the economics of biodiesel. <p>Methanol is the most common alcohol used in the transesterification process due to its low cost. However, recently, ethanol has been promoted as an alcohol for use in transesterification since it can be produced from renewable resources such as switchgrass, corn and wood, thereby reducing the dependency on petroleum sources (Pimentel and Patzek, 2005). A mixture of methanol and ethanol is hypothesized to take the advantages of both methanol and ethanol. The present work is focused on the production of biodiesel from fryer grease via transesterification with various mixtures of methanol and ethanol. Also, the kinetics of transesterification from fryer grease was studied. <p> Since fryer grease contains a high concentration of free fatty acids (FFA) (5.6 wt. %) and water (7.3 wt. %), a two-step acid/alkaline transesterification process was used to produce the esters. Sulfuric acid and potassium hydroxide were used as acid and alkaline catalysts, respectively. The methanol to ethanol molar ratio was varied from 3:3 to 5:1, whereas alcohol to oil molar ratio was maintained at 6:1. After the fryer grease was transesterified, all esters met ASTM standard D-6751. The viscosity of these esters ranged from 4.7 to 5.9 mm2/s. The heating value of the esters was approximately 10% less than that of petroleum diesel. The cloud point and pour point were in the range of 1 to -1 oC and -3 to -6 oC, respectively. When the mixed alcohol was used ethyl esters were also formed at a lower concentration along with methyl esters. The dominant fatty acid in fryer grease esters was found to be oleic acid. The lubricity of kerosene fuel was improved by as much as 33 % through the addition of these esters at rates as low as 1 %. <p>For the kinetic study of alkali-catalyzed transesterification of fryer grease, the alcohol to oil molar ratio, the reaction temperature, and the catalyst loading were varied as 6:1, 9:1, 12:1; 30, 40, 50 oC; and 0.5, 1.0, 1.5 %, respectively. The ester concentration was found to rise with an increase in the catalyst loading or the reaction temperature and with a decrease in the alcohol to oil molar ratio. The overall forward and backward reaction orders were assumed to follow first and second order kinetics, respectively. The kinetic parameters were calculated using MATLAB. The conversion of triglyceride to diglyceride was found to be the rate determining step (RDS) of the overall reaction, with an activation energy of 36.9 kJ/mol.

Biofuels are a developing kind of fuel whose origin is biomass. Among them, many different kind of fuels can be found: bioethanol, biobutanol, biodiesel, vegetable oils, biomethanol, pyrolysis oils, biogas, and biohydrogen. This thesis work is focused on the production of biodiesel, which can be used in diesel engines as a substitute for mineral diesel. Biodiesel is obtained from different kinds of oils, both from vegetable and animal sources. However, vegetable oils are preferred because they tend to be liquid at room temperature. The process to obtain biodiesel implies first a reaction between the oil and an alcohol, using a catalyst and then a sedimentation, where the biodiesel and the glycerol, the two products that are obtained, can be separated because of their difference in density. After the separation, raw biodiesel is obtained and a treatment with either water bubbling or dry cleaning products is needed to obtain the product which will be ready to use. Many methods are available for the production of biodiesel, most of them require heat for the transesterification reaction, which converts the oil into biodiesel. Apart from that, in many cases biodiesel is produced by big companies or by individuals but using complicated and expensive installations. This work is an attempt to develop a way of producing biodiesel without any use of external heat, using a simple procedure which could be used by people with a low knowledge of chemistry or chemical processes. It also seeks to set an example on how biodiesel can be easily made by oneself without the use of any industrial systems, with a low budget and limited need of supervision over the process. In order to achieve that, many trials were undertaken, introducing changes in the different parameters that are responsible for the changes in the final product. Among them, changes in the amount and type of catalyst, the way the catalyst is added, the type of oil used, the time of reaction and the temperature were made. Apart from that, different types of cleaning were tried, starting by water cleaning and then using powder type products, Magnesol, D-Sol and Aerogel. A centrifuge was also tried to test its utility when separating impurities from liquids or different liquid phases. The results of the different trials were analysed using various tests, the most important being the 3:27 test, the solubility test, the soap titration and pH measurements. To sum up, it could be said that the investigation was a success, since it was proved that biodiesel can be made without the use of any external heat with both alkali and acid catalysts, as well as with different ways of adding the catalyst. As for the cleaning, good results were obtained with both dry products and water cleaning, since the soap content of the biodiesel was reduced in both cases. Apart from that, the centrifuge proved to be valid to eliminate impurities from raw oil.

Biodiesel is a renewable, biodegradable, environmentally benign fuel for use in the diesel engines. It can be produced from renewable sources such as vegetable oils or animal fats. Although this fuel has gained worldwide recognition for many years, it is not being widely commercialized mainly because it is more expensive than petroleum diesel. A cheaper feedstock, such as fryer grease, may be used to improve the economics of biodiesel. <p>Methanol is the most common alcohol used in the transesterification process due to its low cost. However, recently, ethanol has been promoted as an alcohol for use in transesterification since it can be produced from renewable resources such as switchgrass, corn and wood, thereby reducing the dependency on petroleum sources (Pimentel and Patzek, 2005). A mixture of methanol and ethanol is hypothesized to take the advantages of both methanol and ethanol. The present work is focused on the production of biodiesel from fryer grease via transesterification with various mixtures of methanol and ethanol. Also, the kinetics of transesterification from fryer grease was studied. <p> Since fryer grease contains a high concentration of free fatty acids (FFA) (5.6 wt. %) and water (7.3 wt. %), a two-step acid/alkaline transesterification process was used to produce the esters. Sulfuric acid and potassium hydroxide were used as acid and alkaline catalysts, respectively. The methanol to ethanol molar ratio was varied from 3:3 to 5:1, whereas alcohol to oil molar ratio was maintained at 6:1. After the fryer grease was transesterified, all esters met ASTM standard D-6751. The viscosity of these esters ranged from 4.7 to 5.9 mm2/s. The heating value of the esters was approximately 10% less than that of petroleum diesel. The cloud point and pour point were in the range of 1 to -1 oC and -3 to -6 oC, respectively. When the mixed alcohol was used ethyl esters were also formed at a lower concentration along with methyl esters. The dominant fatty acid in fryer grease esters was found to be oleic acid. The lubricity of kerosene fuel was improved by as much as 33 % through the addition of these esters at rates as low as 1 %. <p>For the kinetic study of alkali-catalyzed transesterification of fryer grease, the alcohol to oil molar ratio, the reaction temperature, and the catalyst loading were varied as 6:1, 9:1, 12:1; 30, 40, 50 oC; and 0.5, 1.0, 1.5 %, respectively. The ester concentration was found to rise with an increase in the catalyst loading or the reaction temperature and with a decrease in the alcohol to oil molar ratio. The overall forward and backward reaction orders were assumed to follow first and second order kinetics, respectively. The kinetic parameters were calculated using MATLAB. The conversion of triglyceride to diglyceride was found to be the rate determining step (RDS) of the overall reaction, with an activation energy of 36.9 kJ/mol.

Abstract The increasing awareness of the depletion of fossil fuel resources and the environmental benefits of biodiesel fuel has made it more attractive in recent times. The cost of biodiesel, however, is the major hurdle to its commercialization in comparison to petroleum-based diesel fuel. The high cost is primarily due to the raw material, mostly neat vegetable oil. Used cooking oil is one of the economical sources for biodiesel production. However, the products formed during frying, can affect the transesterification reaction and the biodiesel properties. This paper attempts to review various technological methods of biodiesel production from used cooking oil. The analytical methods for high quality biodiesel fuel from used cooking oil like GC, TLC, HPLC, GPC and TGA have also been summarized in this paper. In addition, the specifications provided by different countries are presented. The fuel properties of biodiesel fuel from used cooking oil were also reviewed and compared with those of conventional diesel fuel.

The process of transesterification is the exchange of the organic group R” of an ester with the organic group R’of an alcohol, often catalyzed by acid, base or enzyme. Biodiesel, a mixture of monoalkyl esters of long chain fatty acids, is produced from vegetable oils, animal fats and fish oils by transesterification in presence of alcohol. Biodiesel is a fuel which can be used in a mixture of other fuels or alone. The base catalyzed transesterification method of biodiesel production is not suitable for waste animal fat as it contains 10–15% free fatty acids which result in higher soap formation and cause extensive downstream processing. Enzyme catalyzed transesterification can overcome the problem of soap formation and multi-step purification of end products and results in a higher purity biodiesel. Lipase is the enzyme widely used in the process of enzymatic transesterification. Various lipases have been used to transesterify triglycerides with short chain alcohols to alkyl esters. The objectives of this study were to screen lipase enzymes for the transesterification process and to use the best lipase for biodiesel production from waste animal fat. Enzymatic transesterification by individual and combined enzyme catalysts (Novozyme 435 and NS88001) was first carried out to investigate the effects of reaction time (4, 8, 12 and 16 hour), oil : alcohol molar ratios (1:1, 1:2, 1:3, 1:4 and 1:5), the effects of alcohol type (methanol and 2-butanol) and reaction temperature (35, 40, 45 and 50°C) on biodiesel yield in solvent and solvent-free systems. The highest conversion yield of biodiesel (96.67%) was obtained from a combination of Novozyme and NS88001 lipase with the optimal reaction condition of oil : 2-butanol molar ratio of 1:4, enzyme concentration of 25% (12.5% w/w of each enzyme), hexane as solvent, a 45°C reaction temperature, a reaction time of 16 h and a mixing speed of 200 rpm. The reusability of lipase enzymes by individual and combination of enzyme catalysts (Novozyme 435 and NS88001) with solvent and solvent-free systems was also investigated in order to reduce the cost of the process. The lipase enzymes lost their activity after being reused for 30 cycles in solvent-free systems and after 10 cycles in solvent system.

No description available.

The physicochemical characteristics of seven cold-pressed flaxseed oils sold in New Zealand were investigated for their fatty acid composition, tocopherol composition, moisture and volatile matter, free fatty acids, chlorophyll pigments, unsaponifiable matter, total phenolic acids and flavanoids, and colour. The seven cold-pressed flaxseed oils exhibited significant variations in their physicochemical characteristics. Quality of the oils in terms of oxidative stability was also investigated. Four oils were found to be within the limit of good stability oil indices, measured in terms of peroxide value, p-anisidine value, conjugated dienoic acids, specific extinction in ultraviolet spectrum, acid value and food oil sensor readings (to indicate total polar compounds). The role of minor constituents in the oxidative stability of two selected oils with different levels of fatty acid composition and minor constituents was investigated. Pan heating at 150�C caused loss of tocopherols, plastochromanol-8, phenolic acids, chlorophyll pigments, β-carotene and lutein and changes in the fatty acid composition. The pan-heated oils exceeded the limit of good stability oil indices using the measurement mentioned above except for acid value. The addition of α-tocopherol to the oils did not provide enhanced protection to the oils in accelerated aging of oil tests at 60�C. It was most likely that phenolic acids present in the oils played a dominant role in the oxidative stability of the oils. Lipase-catalyzed transesterification of triolein with cinnamic and ferulic acids using a commercially available immobilized lipase B from Candida antarctica (Novozym 435) was conducted to evaluate the antioxidant activity of the lipophilized products as model systems for enhanced protection of unsaturated oil. The lipophilized products were identified using Electrospray Ionization-Mass Spectroscopy (ESI-MS). Separation and isolation of two classes of lipophilized products was also achieved using a solid phase extraction method developed in this study for further investigation into the structure-free radical scavenging activity. Free radical scavenging activity was determined using the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) method. The polarity of the solvents proved important in determining the free radical scavenging activity of the substrates. Ferulic acid showed much higher free radical scavenging activity than cinnamic acid, which had limited activity. The esterification of cinnamic acid and ferulic acid with triolein resulted in significant increase and decrease in the free radical scavenging activity, respectively. These opposite effects were due to the effect of addition of electron-donating alkyl groups on the predominant mechanism of reaction (hydrogen atom transfer or electron transfer) of a species with DPPH. The effect of esterification of cinnamic acid was confirmed using ethyl cinnamate which greatly enhances the free radical scavenging activity. Although, compared with the lipophilized cinnamic acid product, the activity was lower. The free radical scavenging activity of the main component isolated from lipophilized cinnamic acid product using solid phase extraction, monocinnamoyldioleoylglycerol, was as good as the unseparated mixture of lipophilized product. Based on the ratio of a substrate to DPPH concentration, lipophilized ferulic acid was a much more efficient free radical scavenger than lipophilized cinnamic acid. Lipase-catalyzed transesterification of flaxseed oil with cinnamic and ferulic acids using Novozym 435 was conducted to evaluate whether the lipophilized products provided enhanced antioxidant activity in the oil. The lipophilized products were identified using ESI-MS and were examined for their free radical scavenging activity toward DPPH in ethanol and ethyl acetate. Ferulic acid showed the highest free radical scavenging activity among all substrates tested while cinnamic acid had negligible activity. The effect of esterification of cinnamic acid and ferulic acid with flaxseed oil was similar to that with triolein. Lipophilized ferulic acid was a better free radical scavenger as compared with lipophilized cinnamic acid and extended the naturally-occuring antioxidant capacity of the flaxseed oil. Lipophilized cinnamic acid did not provide much enhanced radical scavenging activity in the flaxseed oil as the presence of natural hydrophilic antioxidants in the oil had much greater radical scavenging activity. It may still be useful for unsaturated oils with a small amount of natural antioxidants in them. Lipophilized cinnamic and ferulic acids showed higher free radical scavenging activity when tested in a less polar solvent (ethyl acetate) whereas ferulic acid showed better activity in a more polar solvent (ethanol). These results indicate that the choice of solvent for the DPPH assay is critical in evaluating the free radical scavenging activity of substrates of differing polarity, and support previous observations by other authors that the solubility of an antioxidant in relation to the site of oxidation is an important factor for consideration in the use of antioxidants.

The physicochemical characteristics of seven cold-pressed flaxseed oils sold in New Zealand were investigated for their fatty acid composition, tocopherol composition, moisture and volatile matter, free fatty acids, chlorophyll pigments, unsaponifiable matter, total phenolic acids and flavanoids, and colour. The seven cold-pressed flaxseed oils exhibited significant variations in their physicochemical characteristics. Quality of the oils in terms of oxidative stability was also investigated. Four oils were found to be within the limit of good stability oil indices, measured in terms of peroxide value, p-anisidine value, conjugated dienoic acids, specific extinction in ultraviolet spectrum, acid value and food oil sensor readings (to indicate total polar compounds). The role of minor constituents in the oxidative stability of two selected oils with different levels of fatty acid composition and minor constituents was investigated. Pan heating at 150�C caused loss of tocopherols, plastochromanol-8, phenolic acids, chlorophyll pigments, β-carotene and lutein and changes in the fatty acid composition. The pan-heated oils exceeded the limit of good stability oil indices using the measurement mentioned above except for acid value. The addition of α-tocopherol to the oils did not provide enhanced protection to the oils in accelerated aging of oil tests at 60�C. It was most likely that phenolic acids present in the oils played a dominant role in the oxidative stability of the oils. Lipase-catalyzed transesterification of triolein with cinnamic and ferulic acids using a commercially available immobilized lipase B from Candida antarctica (Novozym 435) was conducted to evaluate the antioxidant activity of the lipophilized products as model systems for enhanced protection of unsaturated oil. The lipophilized products were identified using Electrospray Ionization-Mass Spectroscopy (ESI-MS). Separation and isolation of two classes of lipophilized products was also achieved using a solid phase extraction method developed in this study for further investigation into the structure-free radical scavenging activity. Free radical scavenging activity was determined using the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) method. The polarity of the solvents proved important in determining the free radical scavenging activity of the substrates. Ferulic acid showed much higher free radical scavenging activity than cinnamic acid, which had limited activity. The esterification of cinnamic acid and ferulic acid with triolein resulted in significant increase and decrease in the free radical scavenging activity, respectively. These opposite effects were due to the effect of addition of electron-donating alkyl groups on the predominant mechanism of reaction (hydrogen atom transfer or electron transfer) of a species with DPPH. The effect of esterification of cinnamic acid was confirmed using ethyl cinnamate which greatly enhances the free radical scavenging activity. Although, compared with the lipophilized cinnamic acid product, the activity was lower. The free radical scavenging activity of the main component isolated from lipophilized cinnamic acid product using solid phase extraction, monocinnamoyldioleoylglycerol, was as good as the unseparated mixture of lipophilized product. Based on the ratio of a substrate to DPPH concentration, lipophilized ferulic acid was a much more efficient free radical scavenger than lipophilized cinnamic acid. Lipase-catalyzed transesterification of flaxseed oil with cinnamic and ferulic acids using Novozym 435 was conducted to evaluate whether the lipophilized products provided enhanced antioxidant activity in the oil. The lipophilized products were identified using ESI-MS and were examined for their free radical scavenging activity toward DPPH in ethanol and ethyl acetate. Ferulic acid showed the highest free radical scavenging activity among all substrates tested while cinnamic acid had negligible activity. The effect of esterification of cinnamic acid and ferulic acid with flaxseed oil was similar to that with triolein. Lipophilized ferulic acid was a better free radical scavenger as compared with lipophilized cinnamic acid and extended the naturally-occuring antioxidant capacity of the flaxseed oil. Lipophilized cinnamic acid did not provide much enhanced radical scavenging activity in the flaxseed oil as the presence of natural hydrophilic antioxidants in the oil had much greater radical scavenging activity. It may still be useful for unsaturated oils with a small amount of natural antioxidants in them. Lipophilized cinnamic and ferulic acids showed higher free radical scavenging activity when tested in a less polar solvent (ethyl acetate) whereas ferulic acid showed better activity in a more polar solvent (ethanol). These results indicate that the choice of solvent for the DPPH assay is critical in evaluating the free radical scavenging activity of substrates of differing polarity, and support previous observations by other authors that the solubility of an antioxidant in relation to the site of oxidation is an important factor for consideration in the use of antioxidants.