1. EXECUTIVE SUMMARY

The focus of the paper is the examination of technology to use the algae Chlorella vulgaris as a feedstock for algal oil for use in transesterification into biodiesel. The Overview summarizes the part biodiesel plays in the energy equation. It also touches on alternative technology which uses biomass to produce liquid biofuels. The Literature Survey reviews some of the emerging science relative to use of enzymes to catalyze bio-oils into biofuels.

Alternative Biofuels mentions the touches on methods used to extract biofuel from biomass using such technologies as direct enzymatic catalysis conversation, thermal depolymyrization. Evaluation of Chlorella as a source of algal oil ties the earlier paper on the commercial production of algae for food and fuel by James E. Miller to this study. Separation of parent and daughter cells examines this need and brings to bear filtration technology which will be deployed first at the rotating harvest cylinder and secondly using a vibratory membrane separator. Enzymatic catalysis is a forthcoming area of technology which must be closely watched since it offers the prospect of production of biodiesel directly from the algal oil, thus saving considerably on the costs of production.

A deeper look at the enzymatic catalysis process lays the foundation for further studies into this field, currently being led by the researchers at Rice University, Houston, Texas. Cell wall rupture is a critical step in the process of extracting lipids and then reusing the cell walls for fermentation into ethanol which is then introduced into the transesterification process. Filtration and polishing of the raw biodiesel is necessary to meet exacting standards of ASTM D 6751, the tests required by Federal and state regulations in order to sell B100 and collect the subsidies and enjoy the tax credits.

This paper examines the use of waste vegetable oil as one of the current sources of feedstock and as the alternative to algal oil. The design of the biodiesel refinery work well for both feedstocks. The design of the algal oil extractor is omitted from this paper because the choices of cell disruption have not been sufficiently research or sourced as an off-the-shelf product. Many investigations remain to be conducted, as suggested by the items listed in paragraph 12. Constructive comments are invited to the email address listed at the conclusion of the main text of the paper.

2. OVERVIEW

The heightened interest in renewable, alternative fuels has driven economic and scientific interest in developing biological oil sources other than oil from oil seeds. ii While the world supply of palm oil and other nut tree oils continue to increase, the focus in the United States and Canada has historically been on oil seeds such as rapeseed, canola, camellia, mustard, and soy. Ethanol has made substantial gains in its use as a fuel and hence, corn acreage has greatly increased as has the price of corn. Conversion efficiencies also play a part in the energy equation. In terms of efficient energy production and overall carbon dioxide emissions, biodiesel trumps not only conventional diesel, but also gasoline and gasoline’s bio-fuel substitute, ethanol. For example, according to a study by the Minnesota Department of Agriculture, biodiesel produces 3.2 units of energy for every unit of fossil-fuel-energy consumed in its production. Ethanol yields a lower 1.34 units of energy, while gasoline and conventional diesel represent a negative yield. (The influx of solar energy into the organic resources—corn, soy, etc.—that will eventually become ethanol or biodiesel accounts for their positive energy yield.) The efficiency of the four fuels breaks down as follows:

FUEL	ENERGY YIELD*	NET ENERGY (LOSS) OR GAIN
Biodiesel**	3.20	220 percent
Ethanol	1.34	34 percent
Petrodiesel	.843	(15.7 percent)
Gasoline	.805	(19.5 percent)

Oil seed and corn crops use of fossil fuel and thus have a large carbon footprint. The issue is how much “work” can the end product accomplish compared to the amount of fossil fuel use in the production of the fuel. Work is defined in terms of heat equivalents. The ratio for soy oil as a feedstock is 1 unit of heat from fossil fuel to 3.2 units of heat from biodiesel. In contrast, the estimated ratio for biodiesel produced from algal oil as a feedstock is in the range of 1: 4 to 5. Much of the science involving algal oil was performed by the National Renewable Energy Laboratory of the U. S. Department of Energy over 20 plus years, consuming over $25 million dollars. Much of the science dealt with the nature of algae and how they can be grown with a high level of lipids. Chlorella vulgaris was one of the leading candidates. iii

After the algal oil program was shut down by Congress in the mid-1990, the approximately 300 “successful” cultivars were delivered to the University of Hawaii, along with the histories of each cultivar. Little effort was made by NREL to design and test the commercial application of the technology. Of the two attempts, the one in Roswell, NM, was most notable as a “failure”. The uncovered pond was in the shape of a race track, with paddles to move the water. This design suffered substantial heat loss when the sun set and during the winter, thus impairing the yield. The authors of the Closing Report recommended that for biodiesel to be successfully produced, petro-diesel would have to rise above $2.00 per gallon.

We are well beyond that point. Earthrise, a highly successful operation producing Spirulina for the health food and supplement market uses open ponds in Calipatria, Imperial County, CA. See: http://earthrise.com/home.asp. Responding to the demand for biodiesel, some 165 plants have been built in the United States, all but two, use transesterification of vegetable oil from oil seeds or recovered waste vegetable oil. iv This process uses alcohol (typically methanol – a petroleum product). Two plants use thermal depolymyrization as a means of converting animal fats and other wastes to a fuel source. The science of algal oil extraction is taking center stage in the development of biofuel. The need for scientific innovation and engineering excellence will become evident. This paper is cast in that light – pushing the envelope to develop the science and engineering necessary for low cost production of algae oil and the refining of such oil into biodiesel. A good history of biodiesel is set forth in Wikipedia. v

3. LITERATURE SURVEY AND LINES OF INQUIRY

The seminal study of algal oil was performed under contract between the National Renewable Energy Laboratory of U. S. Department of Energy. The study was the entry point for serious study of the production of algae for rich content of lipids. vi Much of this paper has benefited from prior work by this author and others. Commercial production of Chlorella vulgaris for its lipid content was the subject of the research paper: Montana Synergy, LLP, Business Plan For Commercial Production Of Algae For Food And Fuel Using Photobioreactor Technology. Published at: http://montanasynergy.wetpaint.com/page/BUSINESS+PLAN+FOR+SPIRULINA+CULTURE+AND+PRODUCTION.


During my investigation of the pathogens of Chlorella vulgaris, background literature vii mentioned the synergistic relationship existed between the protozoa, Paramecium bursaria, and the algae, Chlorella vulgaris. P. bursaria “farms” the Chlorella, extracting nutrients and provides protection to the Chlorella. However, when P. busaria lacks sufficient nutrient, it digests some of the Chlorella. This process suggests the presence of an enzyme capable of digesting the cell walls of Chlorella. This line of inquiry could well lead to the identity of this digestion process which then can be adapted to the dissolution of the Chlorella cell when in the process of producing algal oil.

Another promising avenue of inquiry is the use of enzymes which can directly “snip” the long carbon chains of the polysaccharides from the glycerol molecule. Using this process, scientists at Tsinghua University, Beijing, Peoples Republic of China, have completed pilot tests of an enzyme process which acts on vegetable oil and directly produces biodiesel. viii

If Paramecium Bursaria catalyzes Chlorella by using enzymes, one can assume the enzyme(s) operates on the cell walls of Chlorella by using one enzyme, then by catalyzing the lipids with a different enzyme, it can change the carbohydrates and lipids in to nutrients. One can theorize that the second enzyme “snips” the long chain carbon esters from the glycerol molecule. If these approaches are true, then we will have found the enzymes which directly convert vegetable oil to biodiesel. We would not have to deal with the process discovered by the scientists at Tsinghua University. The approach could drive high net profits as opposed to other systems. Currently transesterification of bio-oils requires alcohol, a catalyst, titration, and heat to make the reaction take place. The raw biodiesel then has to be washed with water, then the water and alcohol extracted. Testing according to ASTM D6751, which follows, is time consuming and expensive.

4. ALTERNATIVE BIOFUELS.

Several promising alternative biofuel sources have been developed. Thermal depolymerzation of animal parts took a leap forward by the construction of a thermal depolyermerization plant by ConAgra next to its Butterball turkey processing plant in Carthage, MO. ix In addition to the other products; the added biodiesel operation produces about 6000 barrels of biodiesel per day from the rendered turkey parts. x

Hydrogen fuel could replace fossil fuel, coal, natural gas, and biofuel. Local production which meets local needs would greatly reduce transportation costs. National Science Foundation teamed with Microbial Electrolysis Cell (MEC) to develop hydrogen production process which uses bacteria acting on biomass to extract the hydrogen. Yields of 91% were reported from vinegar and 68% from trash cellulose.

Bruce Logan of MEC sees a bright future for MEC which has lighted the way toward large-scale production of hydrogen. The papers were published in the November 12, 2007, issue of the online Proceedings of the National Academy of Science; Microbes Churn Out Hydrogen At Record Rate. xi Production of hydrogen fuel is a subject of intense interest by biotechnology engineers. Enzyme catalysis is currently under going rigorous investigation. xii

Other approaches to fermentation by use of enzymes are the use of succinic acid. xiii This approach is a promising line of inquiry. The most likely solution in terms of long-term goals is to use enzyme catalysis to directly convert vegetable oils to biodiesel. However, the time line may be greatly stretched while the scientists at Tsinghua University bring their technology to the market. There are also the legal, political and economic issues associated with these technologies. Enzymic fermentation of the cell walls of Chlorella appears to have the most beneficial result and promise of technology transfer.

5. EVALUATION OF CHLORELLA AS A SOURCE OF ALGAL OIL

Partial evaluation of Chlorella as a candidate for a source of algal oil was made as part of the Business Plan for Commercial Production of Algae for Food and Fuel cited above. The findings reported by that paper were largely taken from the research previously performed under NREL auspices. Subsequent investigation suggests that the choice of this small, spheroid, single cell algae could pose engineering problems. Chief among these problems is the means by which the cell walls can be split or dissolved and the cytoplasm spilled, along with the globules of lipids. In a discussion with Dr. Ganti Murthy, Professor, Oregon State University, it was suggested that enzyme fermentation could lead to the dissolving of the cell walls.

6. SEPARATION OF PARENT AND DAUGHTER CELLS

The algae are harvested by the rotating cylinder described in the above paper and then pumped to the algal oil processing station. The first process before cell wall disruption is to separate the small daughter cells (recently divided – 10 micrometers) from the larger parent cells (25 micometers). Some studies have indicated a size ranging from 10 to 25 micrometers. xiv One can assume that the smaller size represents the daughter cells and the larger size represents the mature cell. xv One of the problems with Chlorella while growing is that some of the cells are mature and thus full sized and some are juvenile and thus smaller. These differences can be seen in the microphoto under End note 1. We want to preserve the daughter cells which are returned to the head of the growing ponds. These cells then continue to propagate the Chlorella cells. To accomplish this process, we need to have a two step process. The first one is to screen the mature cells and allow the juvenile cells to pass through and be returned to the head of the pond. This pass is followed by a second pass which removes most of the water before the mature cells are ruptured. Membrane technology appears to be an appropriate technology to use in this process. \

7. THE ENZYMATIC PROCESS

Use of enzymes will become a major component in the commercial production of biodiesel. The goal of this use is to remove the use of alcohol and catalyst from the process, resulting in a major savings of cost and time. xvi The study of this process is an important part of the future development of algal oil technology. The promise is that it will greatly reduce the cost of degrading the algal cell walls and the refining of the algal oil. To “catalyze” is to accelerate a chemical or biological reaction. The reactants are “substrates” which is a molecule upon which the enzyme is about to operate, and the “product” which is also a substrate, which can be in its final form or in transition to another chemical. Enzymes reduce the energy of activation of chemicals in transition and thus greatly increases the speed of the transition of the chemical reactions.

Enzymes act specifically on substrates. When they act, they do not change the products. The speed of operations is affected by the environment: temperature, pH, concentrations of substrates, and other co-enzymes. Also, inhibitors and accelerators can affect whether or not the enzyme operates and the relative speed of the activation. Enzymes can catalyze millions of reactions per second. For instance, orotidine 5’-phosphate will consume half of its substrate in 78 million years. By adding decarboxylase, the same process takes 25 milliseconds. Enzymes are proteins and range in size from monomers such as 4-oxalocrotonate tautomerase to over 2500 residues in the animal fatty acid synthbase. Enzymes are much larger than the substrates upon which they operate. The region in which the reaction takes places is the “active site”. These proteins are long, linear chains of amino acids which fold to produce three dimensional products, which produce unique structures. The size and shape thus determine the type of binding offered the substrate. The matching of a substrate is not rigid. The substrate molecule is molded as it enters the active site until it is completely bound and its final shape fixed.

Enzymes can act in several ways:

• Lowering the activation energy, thus reducing the amount of energy required for transition and promoting the stability of the substrate/product molecules.
• Lower the energy of the transition site by creating an environment with the opposite charge distribution to that of the transition state.
• Providing an alternative pathway by reacting with a substrate to form an intermediate EX complex which would be impossible in the absence of the enzyme.
• Reducing the reaction entropy change by bringing substrates together in the correct atomic/molecular orientation. Apparently the orientation is stabilized by electrostatic effect by having relatively fixed polar environments.

The dynamics and functions of enzymic reactions provide for regulation of the transition state. Enzymic reactions have different speeds. Initially, the reactions are driven by an abundance of substrates and many enzymes. In interesting animation of the enzymic reactions is shown by a U.C. San Diego reference. xvii Other enzymes such as DNA polymerase produce reactions in a first step and then in a second step, “proof-read” the product for errors. This process has an average error rate of 1 in 100 million reactions in high-fidelity mammalian polymerases.

Enzymes catalyze the forward and backward reactions equally, thus affecting only the speed of the reaction, not the equilibrium. Different enzymes play parts in the processing of foods. Amylases and proteases break down large starches and proteins into smaller ones so that they can be absorbed by the intestines. In herbivores, digestive bacteria in the gut produce cellulase to break down cellulose walls of plant fibers. Enzymes often work together in series, such that the product from one transition reaction becomes the input substrate of the next transition. Network of metabolic pathways within cells depend on functional enzymes which are present. Enzyme activity is controlled (enhanced or diminished) in a cell by a response to the cell’s environment. This effect is called enzyme induction and inhibition. Enzymes can also be “compartmentalized” with different metabolic pathways occurring and used by different parts of a cell such as cytosol, endoplasmic reticulum and the Golgi apparatus. The transitions are looped in the sense that transaction inhibitor is issued as part of the final product, which inhibits the ignition of the process. As the concentration of the product increases, the inhibitor slows the speed of the reaction until the catalytic procedure stops. Post-transitional modification also helps regulate the reactions, such that the reaction is shut down to prevent too high a count of the new product being formed. Enzymes can also be relocated to a different environment. For example, “Chymotrypsin, a digestive protease, is produced in inactive form as a chymotrypsinogen in the pancreas and transported in this form to the stomach where it is activated. This stops the enzyme from disgusting the pancreas and other tissues before it enters the gut. This type of inactive precursor to an enzyme is known as a zymogen.” xviii Enzymes have a vital role in industrial applications. For instance in the baking industry, fungal alpha-amylase enzymes catalyze and breakdown starch in flour to sugar. The yeast action produces carbon dioxide, useful in white bread, buns and rolls. High baking temperatures destroy the yeast. In the biofuel industry, amylases, xylanases, cellulases and lingninases are used. Cellulases are used to breakdown cellulose into sugar which can be fermented into ethanol. xix

8. ENZYME CATALYSIS

“Bonding strain” affects the reaction rate. The induced fit by action of the enzyme at the active site as the substrate enters the active site, induces a structural arrangement which strain the substrate-enzyme bonds so that the energy difference between the unenzymic substrate state and the transition state are reduced, thus lowering the energy difference and helping the catalyzation of the reaction. An animation ably demon-striates this process: http://www.hillstrath.on.ca/moffatt/bio3a/digestive/enzanim.htm Acids and bases, as proton donors and acceptors may donate protons in order to stabilize changes in the transition state. Stabilization can also be affected by the residues in the active site forming ionic bonds with intermediaries. Covalent catalysis forming a covalent bond with the residues helps reduce the energy of activation. This bond is broken to regenerate the enzyme. The rate of reaction is also influenced by the proximity and orientation. The reactions align reactive chemical groups and hold them closer together. These orientations make reactions such as ligations and additions more favorable thus giving a massive rate increase. xx

9. CELL WALL RUPTURE

While technology advances our understanding of the use of enzymes to ferment Chlorella cell walls, xxi here are more traditional means of breaching the cell wall. Use of low pressure flash expansion has the positive attributes of low heat and fast reaction (3 seconds). This process heats the mature cells to the point where the water inside the cells begins to form steam. Quickly reducing the pressure causes the steam to break through the cell walls, turning the cell walls in to a “cup” shape. xxii A general primer on cell wall disruption is covered by Wikipedia. This source advises: “For cells that are more difficult to disrupt, such as bacteria, yeast, and algae, hypotonic shock alone generally is insufficient to open the cell and stronger methods must be used, due to the presence of cell walls that must be broken to allow access to intracellular components” xxiii These stronger methods are discussed below. The summary list for different methods listed by the website is: Laboratory-scale methods, Enzymatic method, Bead method, Sonication, Detergent methods, The 'cell bomb', High-shear mechanical methods, Rotor-stator Processors, Valve-type processors, and Fixed-geometry fluid processors xxiv The commercially sized units are generally those of the high-shear processor. High shear units are commonly used in the dairy industry to homogenize milk. xxv Ultrasonic cavitation is also a useful process to breach the cell walls of algae. That process is generally suitable only for small samples such as use in a laboratory. xxvi Specially designed pumps can also deliver high shear values which could create the vortexes and cavitation necessary to rupture the cell walls. The issue before us is that the cell wall of Chlorella vulgaris is a tough, fibrous material, not easily broken. Until bench tests prove otherwise, the most likely process for mass processing is that of the "cell bomb" indicated above, which process is specific to Chlorella. However, since that process is not "off the shelf" the high shear processor is my choice since it is off-the-shelf. Further investigation into the structure and function of enzymes and their operations is published on the Web. xxvii

10. FILTRATION

After the cell walls are broken using either fermentation or high shear processing, the cell walls are immersed in the mix of water, contents of the cells and cell walls. In order to separate the particulate matter from the liquids, a filtration system must process the “soup”. Several methods are suggested: mechanical (poly filter meshes), centrifugal separation xxviii and membrane technology. The microsize presents problems with the first two methods. Membrane separation seems to hold the most promise for large scale for both batch and continuous processing. The best technology is vibratory membrane separation found thus far is from New Logic Technologies. xxix After the cells are ruptured, we then process this “soup” by using membrane technology adjusted to separate the liquids from the broken cells. The broken cell walls are retained on the input (proximal) side of the membrane while the liquid passes to the outlet (distal) side of the membrane. The membrane is periodically “wiped” so that the micro holes in the membrane do not become plugged. If the fermentation method is used, that process goes from the point where enzymic catalysis has broken 99% of the mature cell walls and next uses the vibratory membrane filter to separate out the cell walls. These walls are then pumped to a fermentation process (which is outside the scope of this paper). After ethanol is produced by that process it can be directed to the transesterification of the algal oil into biodiesel. Any waste products from the fermentation process can be used as animal feed (if found suitable) or added to composting operations.

11. POLISHING OF RAW ALGAL OIL

The liquid passing through the wiped membrane separator will contain micro-particles, alga oil, and water. This “crude” algal oil will need further processing which will remove all but the algal oil. The process which offers the best solution is to use a membrane with pores of about 5 microns. After this process, the liquid will be free of particles, but will still contain water. The main problem with all membrane separators is fouling. Wiped surfaces attempt to solve this problem by removing the boundary layer from the input face of the membrane by using a squeegee. A new system using vibrations offers a better solution to filter out particulate matter. The water can be removed by passing it through a large quality of Amberlite medium. The media is about 1/8th inch in diameter and can absorb about 20% of its weight in water. The system is simple enough. A tube containing the small spheroids is built with an inlet and outlet and a screen at the bottom to retain the Amberlite. When the Amberlite has absorbed water to near capacity, it is removed and dried in an oven, then reused. This system should reduce the water content of the algal oil to about .05 percent. xxx Unless the feedstock algal oil is denatured, a remaining water molecule will render four molecules of KOH ineffective. Both the oil and the ethoxide need to have very low water content.

12. DESIGN OF ALGAL OIL EXPELLER/CONCENTRATOR

Rendering Chlorella into algal oil is a multi-step process. Paragraph 7 above lays the foundation for rupture of the cell wall using the most available method of high shear processing.

1. Tank B and heater. The heater in Tank B will bring the temperature of the load to 120o F and will allow for great separation of particulate matter and free fatty acids. The heat is supplied to an internal heat exchange (plate type) inside Tank B by #5D, which is a waste oil burner which uses waste oil as fuel. xxxi
2. Initial WVO separation by filter. A three stage filter (#4) receives the hot oil and successively passes it through the poly filters in decreasing size meshes. Filter A is 100 microns, filter B is 50 microns and filter C is 25 micros. xxxii

General separation technology: Chemical Processing – Digital Edition, http://www.chemicalprocessing.com/, accessed 11-18-2007; Filtration: http://chemicalprocessing.com/resource_centers/separations_technology/filtration.html; Drying: http://chemicalprocessing.com/resource_centers/separations_technology/drying.html

3. Reduction of Fry Oil Contaminants.

Industrial fry oils produce contaminants by heat acting on the oil and products being fried. These impurities reduce the yield of finished biodiesel.. Heat, air, and moisture lead to polymerization, oxidation and hydrolysis. Researchers in Malaysia, using waste palm fry oil studied four absorbants: silica gel, activated carbon, aluminium oxide and acid-activated spent bleaching earth. xxxiii The advantage of algal oil is that such oil will not contain these impurities The chief contaminants are Free Fatty Acids (FFA). Left in the feedstock, FFAAs cause low yield and lead to the formation of soaps using NaOH as the catalyst. Pretreatment of WVO using silica gel decreases the FFAs by 33%, reduction of peroxide by 65%, changing the color from dark brown to light yellow, and lowered the viscosity from 46.5 cST at 40 oC to 29.2 cSt at 40 oC. The treated oil was less stable from the pre-treated oil due to the loss of almost all of the oil’s antioxidants; this effect changed the induction period from 24 hours to 3.9 hours. The composite effect of the silica gel treatment was to yield 80% methyl esters with only 1% monoglycerides.

4. Degumming. [Future project]
5. Titration. [Future project]
6. Ethoxide. [Future project]
7. Wash water. [Future project]
8. Ultrasonic mixing.

Ultrasonic mixers are employed to better mix the ethoxide, catalyst and oil. Heat is applied to raise the molecular activity and thereby cause more exposure of the triglcerides to the catalysis by KOH. The activity of the ultrasonic reactors is to greatly reduce the aggregation of oil and thereby increase the availability of bonding (active) sites of the substrates to the KOH. The net effect of this proven procedure is to reduce the time for the glycerol to settle out from eight hours to two hours. xxxiv

9. Aeration mixing.

“A unique method of aeration mixing was discovered. If air is introduced deep into the water layer through a sandstone, glass or stainless steel gas diffusion disk numerous air bubbles are formed in the water phase. These numerous water coated bubbles rise through the liquid interface into the ester, carrying large amounts of water in the film, and accomplishing washing as they rise up through the ester. Upon reaching the surface, the bubbles burst and form droplets of water which fall back down through the ester, further washing it. These bubbles and droplets seem to be of such size and nature that the droplets formed do not remain emulsified when they reach the aqueous phase, but quickly coalesce and disappear into the aqueous layer. This method greatly magnifies the interface area, and at the proper aeration rate, half or more of the ester phase volume seems to be filled with quite rapidly settling droplets of aqueous phase.” xxxv Vendors are listed in the end notes. xxxvi

10. Waste oil collection.

Waste fry oil from local restaurants and commercial frying operations will be selected and collected on a regular basis. The small restaurants produce about 40 gallons a week with larger institutions producing 100 to 120 gallons a week. There is generally no fee paid for such collected oil since most smaller restaurants pay to have the waste oil hauled away. The vegetable oil from the fryers typically are dumped into 5 gallon bucket and then carried to the oil dumpster, lifted and poured into the hatch. Our collection system will be two, 55 gallons drums, laid on their sides. Oil will be dumped into a funnel which is plumbed into both drums. Generally, this oil will be very cold, especially in the winter. The system is designed so that hot oil from the tank on the truck will be pumped into each drum, then stirred with a portable electric mixer, then removed by a vacuum pump mounted on the truck and deposited in the tank on the truck. This system allows for collection at times convenient to our schedule and allows oil to accumulate so that our trips achieve the maximum amount of oil from the services to the restaurants. Generally, the pickup will be on a weekly basis for most smaller and medium restaurants. Larger operations may required two or more waste platforms or more frequent pickup. The objective is to maintain a sustainable level of production at the refinery on a predictable cycle. The intent is to allow the employee of a restaurant to deliver the warm oil directly to the platform at a level of lower than the normal waste oil hauler. We will probably need a waste oil hauler permit issued by Oregon DEQ, unless we can fit within an exemption. Attached are the drawings of the WVO Collection Platform. Plan for WVO Collection Platform xxxvii Perspective view of WVO Colleciton Platform xxxviii Funnel xxxix

13. TESTING.

Testing of biodiesel is mandated by U.S. EPA, IRS, DOT and DOE for compliance with ASTM D6751. This test protocol is a panel of tests designed to test petrodiesel, some of which are made applicable to testing biodiesel. Although these tests are normal for large petrodiesel plants, they pose an impossible problem for the small producer. The tests must be performed on each “batch”. The cost of the entire panel costs between $500 and $850, which puts testing outside of all possible profits. There are other equivalent testing, namely use of near-infra red spectrometry. Kits are offered by some vendors to test diesel such as by Herguth Laboraories, Inc. http://www.herguth.com/?gclid=CNa00e-r548CFSW-YAodHgMCjg, accessed 11-25-2007. California Air Quality regulations permit the sale of “development fuels” which do not meet ASTM D6751 to be sold under a variance. xl

14. CONCLUSION

This paper covered the study of the algae, Chlorella vulgaris, as a primary candidate for use in the commercial production of algal oil as a feedstock for biodiesel production. There are many emerging levels of technology which will change the choice of feedstock as well as the manner in which the lipids are extracted. Not covered is the considerable scientific interest in genetic engineering of selected species of lipid rich algae. There is current interest is the selection of a enzymic catalysis process which has the promise of reducing or eliminating the use of alcohol and catalyst for transesterification and the use of water to wash the impurities from the raw biodiesel. This study also brought the science of algal oil production into the design stage whereby a refinery was designed using off-the-shelf equipment for the transesterification process. Also, a system to degrade the cell walls of Chlorella and extract the algal oil was designed such at the daughter cells were saved and routed back to the growing ponds and the mature cells processed for their oil content. The system also separates the disrupted cell walls for fermentation to produce ethanol which is required for the biodiesel refinery. There remain several areas of future study, noted in the paper. Also, this paper is a prelude to a business plan which will be based on the science and engineering presented by this paper.

Respectfully submitted,

James E. Miller, BA, B.S., J.D.
Email: jimmiller5417@yahoo.com [preferred] Cell: 541-971-0403


END NOTES
i
Yusuf, Christi, Biodiesel from Microalgae, Biotechnology Advances, 25, (2007), 294-306, Institute of Technology and Engineering, Massey University, Private Bag 11 222, Palmerston North, New Zeeland, 12 February, 2007, www.elsevir.com/locate/biotechadv

Abstract