La importancia de los peces fluorescentes (Spanish Edition)

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Click here for the english version. For other languages click here. Grandes agrupaciones de embriones pueden ser generados y disponibles semanal para las necesidades experimentales. Please recommend JoVE to your librarian. A Embriones de pez cebra en 72 HPF fueron sometidosa heridas como se describe anteriormente por Niethammer et al. La barra de escala 20 micras. La barra de escala, m. E00 Voltaje , 2,1 AmpGain , Modo: Una muestra no incubada con la sonda ROS-sensible se ha incluido como control negativo. FACS acervoajustes ition fueron los siguientes: Las mediciones se relacionan con muestras mostradas en C.

Una vez que la sonda fluorescente se oxida por ROS no pueda entrar de nuevo a su no fluorescente estado reducido. Los principales componentes de este grupo son: Really it's interesting paper, But please Can we Know the ROS-sensitive prob that used in this paper in case of generic and mitochondrial oxidative stress? You must be signed in to post a comment. Please sign in or create an account. Skip to content Biology. La barra de escala, m. E00 Voltaje , 2,1 AmpGain , Modo: Una muestra no incubada con la sonda ROS-sensible se ha incluido como control negativo. FACS acervoajustes ition fueron los siguientes: Las mediciones se relacionan con muestras mostradas en C.

Una vez que la sonda fluorescente se oxida por ROS no pueda entrar de nuevo a su no fluorescente estado reducido. Los principales componentes de este grupo son: Really it's interesting paper, But please Can we Know the ROS-sensitive prob that used in this paper in case of generic and mitochondrial oxidative stress?

You must be signed in to post a comment. Please sign in or create an account. Skip to content Biology. Your institution must subscribe to JoVE's Biology section to access this content. Fill out the form below to receive a free trial or learn more about access: Autoclave listo para el uso del agua de pescado antes de su uso.

Centrifugar la metilcelulosa a xg durante 5 min antes de usar. Use de acuerdo con las pautas de manejo apropiadas. El contacto con material combustible puede causar fuego. En la Figura 7. De nuevo, la cantidad de contenido de aceite crudo en la semilla depende de la calidad de la semilla. Por lo tanto, para garantizar la consistencia de los productos, es esencial disponer de materias primas consistentes. El metanol y el agua se reciclan, lo que da como resultado el producto: Also the gums and other impurities should be removed from the oil for better product yield and quality.

Therefore, it is essential that oil should be pre-treated to meet above limit. The pre-treatment of the oil is carried out in the preparatory section. Bleaching is carried out to lighten the colour of the oil. Following sections are used for production of biodiesel from the oil. Crude non-edible oil is the usual feed stock eg. The oils have impurities which interfere with the process of trans-esterification and must be removed before the process in a Preparatory step. Then soap stock formed due to chemical reaction is allowed to settle at the bottom of the neutralizer from where it is taken out into soap pan, as soap will complicate the separation process.

The free fatty acids are distilled out and condensed separately for further conversion to Methyl-ester. Bleaching Neutralized oil is drawn into the second vessel called bleacher where colour of oil is removed by bleaching process with aid of chemicals such as carbon black and bleaching earth. Stirring is also continued. Bleaching process is done under vacuum.

Bleached oil then goes to the filter press where bleaching earth and chemicals are separated. The refined oil if meeting the following standards shall be introduced to the transesterification section. Tranesterification Section The transesterification process flow chosen is similar to a cross flow. The feedstock and the intermediate product, respectively, flow successively through the two mixer-settler units, both units being supplied with methanol and catalyst in parallel.

The reactors are of multi-stage design, comprising various reaction chambers in order to achieve maximum conversion efficiency to methyl ester. The settlers allow phase separation to reach virtually the limit of solubility. This is of great benefit for the downstream processing of the phases rich in ester and glycerin.

This process configuration is supported by a defined temperature profile adjusted along the mixer-settler arrangement. The ester-rich phase still contains methanol, glycerin, small amounts of catalyst, soaps and high boiling components. In order to remove the water-soluble substances, the ester phase is washed. To avoid the formation of emulsions, any soap that may be present is previously split by adding acid.

The washed ester still contains some water, being vacuum-dried in a dryer circuit to adjust the tolerable water content. The high-glycerin phase is collected in a pump tank together with the wash water because of its methanol and glycerin contents. In this process, the catalyst decomposes into caustic soda solution and methanol in the presence of water. The catalyst in liquid form is added to the methanol by means of metering pumps. As the catalyst is sensitive to water, the catalyst pump tank may be protected with a nitrogen layer.

Diluted hydrochloric acid is prepared automatically and can be added to the methyl ester to be washed. All gases vented off from the mixer-settlers, from the catalyst pump tank and the methanol pump tank are routed to a condenser where they are partly condensed and returned to the process in liquid form. The methanol containing phases e. By means of a pump, the dried methyl ester is continuously delivered to the methyl ester storage tank provided by customer.

The free fatty acids present in vegetable oils are eliminated in the preparatory section mentioned above. The high-free-fatty acid feedstock is mixed with alcohol and acid in a separate reactor, converting all free fatty acids into methyl esters. The evaporator system has two re-circulating evaporation circuits stages maintained under vacuum; each re-circulating evaporation circuit is composed of a heat exchanger and evaporator section.

The vapors from the last evaporator stage are precipitated in a condenser. The concentrated glycerol solution is transferred to a storage tank for storage and sale. This operation is only required if the project is intent on capturing this additional value at the expense of significantly higher capital and operating costs. Base catalyzed transesterification with refined oils 2. Base catalyzed transesterification with low fatty acid greases and fats 3.

Acid esterification followed by transesterification of low or high FFA free fatty acid fats and oils Many other processes are being developed, which include the following. Some companies offer pre-made turn-key plants, some will custom build a turn-key plant to suit specific customer needs, while others offer engineering services for a locally built, custom plant. All options have advantages and disadvantages. Most of the high-tech biodiesel plant manufacturing companies offer plants that are economically feasible for centralized, large-scale production using a continuous process.

In order to reach these economies of scale, these plants operate mostly on virgin oils. Decision Making Criteria and Parameters A variety of process options were investigated and evaluated with respect to the volumes and characteristics of feedstock, products and by-products, capital and operating costs, production flexibility, and the ability to meet required finished product specifications on a consistent and reliable basis.

This included an analysis of batch vs. The Company considered the following critical success factors prior to making the final choice for the technology platform: The selection of appropriate technology for production of bio-diesel requires careful selection of processing steps, catalysts, and downstream process integration. The quality of vegetable oils, particularly FFA and moisture content, plays an important role in identifying the technology suitable for a particular situation.

To reduce the cost of bio-diesel production, value-addition options for by-products such as oil cake and glycerine are also being researched. Important factors to be considered for selection of appropriate technology to produce bio-diesel include the following. The proposed Jatropha plantation shall be completed in and the first exploitable harvest shall be available in Therefore, following observations are very essential to be followed: With own Jatropha feed stock, the company shall start the production in first oil extraction and biodiesel plant of 10 tpd capacity in The plant either shall be run on reduced capacity utilization in since seed available for oil extraction shall not be sufficient to run for days in a year.

It is recommended that company may procure feedstock from the market to use both the plants on full capacity. After this the company feedstock shall be sufficient to run both the plants. Since the production of feed stock shall be increased every year it shall be appropriate to install two number of biodiesel plants in a phase manner of 5 tpd capacity. Equipment order will be possible after negotiations with producers of different parts and completing installation plan.

Quantity of agricultural tools needed was estimated for a farm with total arable area 1, ha. Site preparation includes mainly construction works: This will be done by subcontractor. We can start processing of oil right after site preparation, because this does not require complicated technology. So, installation of oil processing equipment oil presses, filters, cisterns and offering oil processing services can be started at the beginning of the year.

Experience in the rapidly growing biodiesel market in Europe shows that the legally demanded level of quality EN is often not achieved due to poor process technology and inadequate production management. As such the quality assurance for biodiesel is very essential since off specification biodiesel shall not only damage the engine parts but also shall be hard to be sold. Sulphated Ash ISO 0. Specific Gravity The specific gravity is a relative measure of the density of a substance. The specific gravity of conventional No. Figure given below shows a hydrometer for measuring specific gravity.

Other methods are also in common us Hydrometer for measuring specific gravity Kinematic Viscosity To define kinematic viscosity it is useful to begin with the definition of viscosity. We intuitively understand viscosity as evidenced by the fact that we know, for example, honey is more viscous than water. There is a hydrodynamic definition of viscosity as well. Technically it is the ratio of the shear stress to the shear rate for a fluid. The test is usually conducted by measuring the time required for a volume of liquid to flow under gravity through a calibrated glass capillary tube.

The kinematic viscosity is then equal to the product of this time and a calibration constant for the tube. Biodiesel is more viscous than No. Flash Point A key property determining the flammability of a fuel is the flash point. The flash point is the lowest temperature at which an applied ignition source will cause the vapors of a sample to ignite. Therefore, it is a measure of the tendency of a sample to form flammable mixture with air.

The flash point is determined by heating a sample of the fuel in a stirred container and passing a flame over the surface of the liquid. If the temperature is at or above the flash point, the vapor will ignite and an easily detectable flash can be observed. Figure below shows the test cup for the apparatus. The cup is filled with fuel and heated with an external heater.

The agitator ensures that the fuel temperature is uniform. A small open flame is maintained from an external supply. Periodically, the stirrer is stopped and the flame is pivoted down to an opening in the top of the cup to see if the fuel vapors will ignite. The ignition source is shown Figure below. When the flash point has been reached, there will be a small flash that is sometimes accompanied by an audible popping sound.

Occasionally, the flash may actually extinguish the flame on the ignition source. For a pure substance the boiling point is a single temperature value. However, for a mixture of hydrocarbons as exists in diesel fuel there is a range of boiling points for the different constituent chemical species. The distillation test is an alternative procedure for determining the volatility characteristics of a petroleum product the distillation characteristics, i. Distillation limits are commonly included in petroleum product specifications due to the number of chemicals species that can be present.

The standard procedure for measuring the boiling point range for diesel fuels is the distillation test, which is ASTM D The simple distillation apparatus used for this test is shown in Figure below. To run the test, a simple batch distillation process is conducted on a sample of the fuel and boiling point data are collected. Higher Cetane numbers indicate shorter times between the injection of the fuel and its ignition.

Higher numbers have been associated with reduced engine roughness and with lower starting temperatures for engines. The cetane number is an engine-based test. It is based on a special engine produced by the Waukesha Engine Company that is similar to the Octane Test Engine used for rating gasolines. The engine is a single cylinder, indirect injection diesel engine.

The combustion chamber is shown in Figure below. Then the engine is switched to operate on a blend of two reference fuels. Different blends are tested until a formulation is found that restores the ignition delay to 13 degrees. The primary reference fuels are n-cetane n-hexadecane , which has a cetane number of and heptamethylnonane HMN , which has a cetane number of When the ignition delay is restored to 13 degrees, the cetane number is computed from the following relationship.

Two ASTM methods are available for computing the cetane index. ASTM standard D gives the following empirical equation for the cetane index: ASTM standard D gives the cetane index according to the following four-variable equation: The cloud point is the temperature at which a cloud of wax crystals first appears in a liquid upon cooling. Operating at temperatures below the cloud point for a diesel fuel can result in fuel filter clogging due to the wax crystals. The apparatus used for this test is shown in Figure below.

The cloud point is an important property for biodiesel since biodiesel fuels typically have higher cloud points, i. This feature has implications on the use of biodiesel in cold weather applications. A summary of the procedure steps is 1 the sample is cooled in a Peltier device at a constant rate, 2 the sample is continuously monitored by optical detectors, and 3 the temperature is recorded that corresponds to the first formation of a cloud in the fuel. The repeatability of the cloud point test is less than0. The pour point is the lowest temperature at which a fuel sample will flow.

The pour point also has implications for the handling of fuels during cold temperatures. Copper Strip Corrosion Detection of copper corrosion from petroleum products by the copper strip tarnish test measures the degree to which the fuel can have a corroding effect on various metals. The corrosivity of a fuel has implications on storage and use of the fuel. Then the strips are washed in a solvent and compared to the descriptions in Table given below. Light orange, almost the same as freshly polished strip b.

Dark orange 2 Moderate Tarnish a. Multicolored with lavender blue or silver, or both, overlaid on claret red d. Brassy or gold 3 Dark Tarnish a. Magenta overcast on brassy strip b. Multicolored with red and green showing peacock , but no gray 4 Corrosion a. Transparent black, dark gray or brown with peacock green barely showing b. Graphite or lusterless black c. Glossy or jet black Ramsbottom Carbon Residue Ramsbottom Carbon Residue of Petroleum Products determines the amount of carbon residue left after evaporation and pyrolysis of an oil, indicating its relative propensity to form coke. It is a potential indicator of the likelihood that a fuel would form deposits from carbon in an engine's combustion chamber.

Heating Value, Net and Gross There are actually two heating values in common use, the higher, or gross, heating value and the lower, or net, heating value. Both quantities are measured using a calorimeter where the heat transfer from the hot gases resulting from combustion of the fuel with air is measured as the gases are cooled to the initial temperature of the reactants. The higher heating value assumes that all of the water in the products is condensed liquid while the lower heating value assumes all of the water is present as vapor, even though the product temperature may be below the dew point temperature.

The lower heating value is the most common value used for engine applications. It is used as an indicator of the energy content of the fuel. In general, the higher the heating value of the fuel, the less fuel that will be required to do a given amount of work. Acid Number Acid number is the quantity of base, expressed as milligrams KOH per gram of sample, required to titrate the sample to a specified end point Biodiesel limit 0.

The acid number is also a measure of free fatty acids. The free fatty acids can lead to corrosion and are a symptom of water in the fuel or fuel oxidation. This is also an indication of the condition of the stability of the fuel, because the acid number increases as the fuel ages.

Test method - The sample is dissolved in a mixture of toluene and iso-propanol that contains a small amount of water. The sample is titrated potentiometrically with alcoholic potassium hydroxide. The meter readings are plotted against the respective volumes of titrating solution and the end points are taken at well defined inflections in the resulting curve. It is applicable to naphthas, distillates, oils, gasoline, diesel, and biodiesel and jet fuel.

Un ensayo para la línea lateral de regeneración en adultos de pez cebra

Some process catalysts used in petroleum and chemical refining can be poisoned when trace amounts of sulfur are present. Sample combustion gases are next exposed to UV light. The SO2 is excited, from which fluorescence is emitted as it returns to its stable state. The fluorescence is detected by a photomultiplier tube and the resulting signal is a measure of the amount of sulfur contained in the sample. Sulfated Ash This test method determines the amount of mineral ash remaining after a fuel is burned. For biodiesel, this test is an important indicator of the quantity of residual metals in the fuel that could come from the catalyst used in the esterification process.

The lower limit of this test is 0. Test Method — The sample is ignited and burned until only ash and carbon remain.

The iodine value is an indicator of the unsaturation of the fuel, which has been linked with formation of engine deposits and problems in storing the fuel. Water and Sediment This test method covers the determination of the volume of free water and sediment in middle distillate fuels.

This test is a measure of cleanliness of the fuel. For B biodiesel it is particularly important because water can react with the esters to make free fatty acids and can support microbial growth in storage tanks. This method used to measure water and sediment is only sensitive to free water. It uses a centrifuge to separate small water droplets and particles to be sure they do not exceed 0.

An earlier method D specified that a solvent should be added to allow the measurement to include dissolved water but since diesel fuel will only dissolve about 50 ppm of water, this has little impact on whether the fuel exceeds the specification value. This test is particularly important when working with biodiesel because biodiesel is usually water-washed to remove traces of soap and free glycerol.

Vacuum drying is usually needed to remove residual water following the washing process. Test Method - A ml sample of undiluted fuel is centrifuged in a tube readable to 0.

La importancia de los peces fluorescentes by Almudena Solana

After centrifugation, the volume of water and sediment which has settled into the tip of the centrifuge tube is read to the nearest 0. Lubricity Lubricity can be defined as: The lower the friction the higher the lubricity. Pure biodiesel and high level blends have excellent lubricity.

Gas Chromatography Chromatographic methods are used to separate a mixture of compounds based on their physical properties. It determines the amount of glycerol in derivatized form , mono- and diacylglycerols both also in derivatized form , triacylglycerols, and methyl esters in a biodiesel sample.

In GC, the mixture is separated mainly by the boiling point and the structure imparting a feature termed polarity of the individual compounds. To carry out a GC analysis, the sample is usually dissolved in low concentrations in an organic solvent and then injected into the gas chromatograph. In some cases, a sample needs to be derivatized with a specific reagent in order to obtain a useful gas chromatogram. This is the case for biodiesel.

Glycerol and the mono- and di-acylglycerols contain free hydroxyl groups, causing these materials not to perform well in GC. Derivatization in the case of biodiesel with a silylating reagent improves their performance considerably. Derivatization can provide better resolution between compounds with similar properties. After injection into the gas chromatograph, the sample is separated on a column. A column is a long, thin path capillary tube that contains a material with which the sample components interact more or less strongly depending on their structure polarity!

Usually, the solvent used and, if applicable, residual derivatizing agent used for preparing the sample are the first materials to elute i. When the detector detects a material eluting from the column at a certain retention time i. Generally, the integrated value of the peak an amplitude over time is proportional to the amount of material causing them.

This constitutes the usefulness of GC in quantifying the amounts of components in a mixture Fuel Stability A fuel is considered unstable when it undergoes chemical changes that produce undesirable consequences such as deposits, acidity, or a bad smell. There are three different types of stability commonly described in the technical literature. Thermal stability addresses fuel changes that occur due to elevated temperature.

These changes may occur at conditions encountered in modern fuel injection systems as fuel is recirculated through the engine cylinder head and back to the fuel tank. Oxidative stability refers to the tendency of fuels to react with oxygen at temperatures near ambient. These reactions are much slower than those that would occur at combustion temperatures, and they produce varnish deposits and sediments.

Storage stability is also a frequently used term and refers to the stability of the fuel while it is in long-term storage. These terms are not necessarily exclusive terms. For example, oxidative attack is probably one of the primary concerns of storage stability but storage stability might also involve issues of water contamination and microbial growth. El aceite crudo no comestible es la materia prima habitual, p. Materia insaponificable, aquella que no se puede saponificar.

La fase alta en glicerina se recolecta en un tanque de la bomba junto con el agua del lavado debido a su contenido de metanol y glicerina. El metanol que contiene fases p. Todas las opciones tienen ventajas y desventajas. Por lo tanto, es sumamente esencial que se sigan las siguientes observaciones: Cenizas sulfatadas ISO 0. Por lo tanto, es una medida de la tendencia de una muestra para formar con el aire una mezcla inflamable. La siguiente figura muestra el recipiente de prueba para el aparato.

El recipiente se llena con combustible y se calienta con un calentador externo.

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El agitador garantiza que la temperatura del combustible sea uniforme. Entonces se hace un cambio para que el motor funcione con una mezcla de dos combustibles de referencia. El aparato usado para esta prueba se muestra en la siguiente figura. El resumen de los pasos del procedimiento es: La corrosividad de un combustible tiene implicaciones en el almacenamiento y uso del combustible.

Color bronce u oro. Magenta nublado en la tira bronceada. Multicolor mostrando rojo y verde como pavo real , pero no gris. Grafito o negro sin brillo. Negro brillante o negro azabache. Esta prueba es una medida de la limpieza del combustible. Lubricidad La lubricidad se puede definir como: Generalmente, el valor integrado del pico y la amplitud en el tiempo, es proporcional a la cantidad del material que los causa. The biodiesel industry has expanded far beyond a niche market to become a significant industrial player. Biofuels are taking on renewed global importance as countries seek to substitute the soaring price of conventional oil and cut hazardous emissions.

Global output of biodiesel is between and million gallons annually. Although Biodiesel can be marketed as an ordinary fuel and our calculations in Financial Planning section are based on market price of conventional diesel, we believe, that most perspective customers are farms, in-city transport companies and those who need lowest possible pollution levels in technological processes: Although with some limitations, this was true for years and seems to be accelerating due to political and economical reasons.

It is necessary to assume, however that because of future increasing of Biodiesel production over the world, prices of Biodiesel can go up only to a sustainable extent. We are part of rapidly growing biofuel business, which includes also producers of heating oils and oleochems. Unless sells of petrodiesel remain subject to excise and other taxes, our product would be uncompetitive on market.

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Biggest problem may appear if state imposes the above-mentioned taxes for Biodiesel, which however, is highly unlikely, because of strategic importance of alternative fuels for country. Strategy and implementation summary Our company believes that qualitative approach will substantially expand the available market for us by creating a new supply route into the mainstream fuels infrastructure that does not exist today: Emphasize inherent qualities of Biodiesel and focus on target markets. We need to establish our business offering as a clear and viable alternative for our target markets.

Strategy pyramid For placing emphasis on quality, our main tactics are technology expertise and developing our own supply network. Our specific programs for technology are continuos learning and research. Specific programs for supply include cultivation of Jatropha seed and cooperative agreements with local farms. This shall be achieved by cost effective scientific farming of Jatropha with proper management. Value proposition We think our value proposition is quite clear and quite easily distinguished from most others in the market.

We offer ecology-friendly and renewable type of fuel at a reasonable market price. Competitive edge Our most important competitive edge is entering Jatropha cultivation and fuel supply industry with newest technologies and gaining experience and market share before competitors came. Unlike mineral fuel, Biodiesel is renewable energy source, with competitive cost and performance. Promotion We depend on direct contacts as our main way to reach new buyers. As we grow, however, we need to change the way we promote ourselves: We'll be developing our core positioning message: We will be using local newspaper advertising, radio, and cable TV to launch the initial campaign.

Distribution We focus on the market need for environment friendly fuel distributed through the authorized gas filling stations. We will also maintain close relationships with transport companies and agricultural enterprises. Sales strategy Our major intention is to keep levels of monthly production, sales, and collections close one to another. Because Biodiesel is highly liquid product, this goal can be implemented with right choice of distributors and suppliers m.

Direct —cost of Biodiesel production is projected under Table: Biodiesel, glycerol and Jatropha cake We must charge appropriately for this unique type of feed stock, but we want also to predict revenues as close as possible and to eliminate market risk, so we plan to it as below Table: Sales programs There are three sales programs in this business plan: Biodiesel is planned to sell using direct mail, seminars, promotion programs and advertising in business newspapers.

Because Biodiesel is a valuable fuel for in-city transportation companies, direct mail to these and other companies would be effective tool in promotion. Ecology, energy, technology related seminars are effective in finding regional representatives, partners, suppliers and buyers of our product. Total budget for all these programs is shown in Profit and Loss Statement, Advertising and Miscellaneous expenses. The seed cake is an excellent fertilizer and has a waste market Glycerin be marketed to costematic, soap industries. Milestones The following table lists 1-st year milestones, with dates and budgets for each.

The milestone schedule indicates our emphasis on planning for implementation. The most important programs are Jatropha cultivation Stages 1 to Harvesting and equipment installation.

We omitted secondary actions like office preparation, stationery purchase or calls to perspective customers. Management Our management philosophy is based on responsibility, mutual respect and entrepreneurial creativity. Our main management divisions are farming, processing, and sales. Farming handles cultivation of Jatropha; processing- pressing of oil seeds and production of Biodiesel. We believe we have a good team for covering the main points of the business plan: Personnel plan The Personnel Plan reflects the need to bolster our capabilities to match positioning in the fuel and agricultural market.

The need for labor force was estimated accordingly to industry experience and technological needs. Project Financing Summary of finance required presented on the table below. We expect outside investor to finance the entire project and we offer him Direct costs were calculated in accordance to market prices of chemicals, all other expenses were also properly allocated in the income statement 6.

This assumption and other financial indicators shown in the table below 8. There are three stages - plantation, extraction, and transesterification - are considered. These stages are considered to be separate entities in this calculation. Cost calculated for the seed is loaded to the extraction stage, assuming that the seed is bought by an extraction unit, and then the corresponding raw oil cost is calculated.

Finally, the raw oil cost is loaded to the transesterification stage, again assuming that a transesterification unit is using this raw oil and transesterifying it to bio-diesel. Unforeseen factors and developments our ability to implement its business strategy may be adversely affected by factors that the we Cannot currently foresee, such as unanticipated costs and expenses, interruptions to or delays in production, reduced demand for the Our product, technological change, loss of political support for Biodiesel or severe economic downturn.

All of these factors may necessitate changes to the business strategy described in this document. Competition There can be no assurance those potential competitors of the company, which may have greater financial, research and development, sales and marketing and personnel resources than ours. It is anticipated that the market will become increasingly competitive over the coming decade. Additionally, several other biofuels and renewable fuels could be introduced to the market as alternatives to mineral diesel. Should these alternatives be selected in the market the demand for Biodiesel could be diluted.

Dependence on key personnel We believe that its future success will greatly depend upon the expertise and continued services of certain key executives and technical personnel, including the executive Directors We cannot guarantee the retention of such key executives and technical personnel. As a result, our business, its results of operations and financial condition may be adversely affected. General risks Our business may be affected by the general risks associated with all companies in the energy industry.

The prices received for our goods and products depend on numerous factors, many of which are beyond its control and the exact effect of which cannot be accurately predicted. Such factors include general economic and political activities, including the extent of governmental regulation and taxation. An investment could be affected adversely by changes in economic, political, administrative, taxation or other regulatory factors The company faces a number of industrial risks such as a dispute with its workforce and dependence on key suppliers, both of which may lead to a deterioration in financial performance.

Risks associated with international sales. Operational risks The financial performance of the Company is at all times subject to operational risks.

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For example, the technology could face unforeseen refining problems, in which case production and financial performance would be delayed. Unanticipated additional maintenance of the machine would also impact the production capacity and revenue projections. Storage tanks should be fully sealed at all times in order to ensure quality Biodiesel and to minimize moisture being added to the fuel. The risk of contamination of the pure vegetable oil or Biodiesel exists from transportation, expelling, storage and refining should the proper quality procedures and handling guidelines not be adhered.

The Company is also at risk of the contamination of its raw materials supplies at source which may lead to stock write downs and an inability to supply customers. The Company attempts to safeguard against this risk by checking materials prior to purchase and dispatch and on receipt at the relevant processing facility. Agricultural risk Our business may be affected by all general risks associated with agricultural production. Theft is also a concern in certain regions, should trees be deemed valuable as another commodity in those regions.

The use of petro-diesel in engines creates a significant amount of greenhouse gas emissions that can be greatly reduced by substituting a blend of Bio diesel and petro-diesel or using percent Bio diesel. Greenhouse gas emissions can be reduced in a number of ways through the Bio diesel production process including: The Bio diesel production process.

Carbon sequestration, seed cake and Bio diesel are potential CER sources while glycerine is a potential additional revenue source. Bio diesel can be produced from plants such as rapeseed, palm, coconut, or soybean or from used cooking oil from restaurants.

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