Source: Agricultural Research Service, Southern Regional Research Ctr submitted to
DEVELOP OF PROCESSES TO IMPROVE OILSEED UTILIZATION
Sponsoring Institution
Agricultural Research Service/USDA
Project Status
TERMINATED
Funding Source
USDA INHOUSE
Reporting Frequency
Annual
Accession No.
0403622
Grant No.
(N/A)
Project No.
6435-41000-082-00D
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Nov 12, 2000
Project End Date
Sep 30, 2004
Grant Year
(N/A)
Project Director
WAN P J
Recipient Organization
Agricultural Research Service, Southern Regional Research Ctr
1100 Robert E. Lee Blvd.
New Orleans,LA 70124-4305
Performing Department
(N/A)
Non Technical Summary
(N/A)
Animal Health Component
(N/A)
Research Effort Categories
Basic
20%
Applied
80%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
5011810200010%
5011810202040%
7121810200010%
7121810202020%
7121899202020%
Knowledge Area
501 - New and Improved Food Processing Technologies; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
1810 - Cottonseed; 1899 - Oilseed and oil crops, general/other;

Field Of Science
2020 - Engineering; 2000 - Chemistry;
Goals / Objectives
To investigate the effect of processing techniques and conditions on the chemical and physical properties of oil, protein and their by-products and on the biologically available gossypol in cottonseed products. To develop innovative separation and purification processes to enhance the utilization of oilseed products with minimum undesirable environmental impact.
Project Methods
The ultimate goal of the project is to develop a process to produce cottonseed products which are consistently safe to lifestock. To achieve this objective, biologically available gossypol in cottonseed products need to be defined and determined using selected animal models and analytical procedures. At the same time, cost effective processes will be developed to extract and deactivate gossypol and aflatoxin, and to enhance value and utilization of its co-products.

Progress 11/12/00 to 09/30/04

Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Because of increasing competition and regulatory pressure, the cottonseed processing industry urgently needs to improve process productivity and product quality, and enhance by-product utilization. Furthermore, the cottonseed oil and meal profit margins have been depressed in recent years due to worldwide over supply of these materials. Cottonseed contains gossypol, a polyphenolic pigment, which limits use of cottonseed products in animal feeds. Over the years, changes in the commercial processing of cottonseed have resulted in different meal products and the safe use of cottonseed products as a feed ingredient is not adequately demonstrated. Determining how these meals can be used optimally should allow for expanded markets for these products. Because of regulatory pressure, alternative and safer extraction solvents are also needed by the industry. At present, the main solvent used for vegetable oil extraction is hexane, which has been classified as a Hazardous Air Pollutants (HAP) by US Environmental Protection Agency (EPA). To assist the oilseed industry, the research team at the Southern Regional Research Center (SRRC) works to improve the utilization of oilseeds through innovative process development. This goal is accomplished through investigating the effect of processing conditions on the chemical and physical properties of oil and co-products, through determining the biological availability of gossypol in cottonseed products and through developing innovative separation and purification processes to enhance process efficiency, improve product quality, and reduce environmental impact. This project is part of National Program 306 - Quality and Utilization of Agricultural Products [New Processes, New Uses, and Value-Added Foods and Biobased Products]. The mission of this program is to enhance the economic viability and competitiveness of U.S. agriculture by maintaining the quality of harvested agricultural commodities or otherwise enhancing their marketability, developing environmentally friendly and efficient processes, and expanding domestic and global market opportunities through the development of value-added food and nonfood products. The program has two specific aims: (1) quality characterization, preservation, and enhancement of agricultural commodities and products and (2) the development of new processes, new uses, and value-added foods and biobased products. 2. List the milestones (indicators of progress) from your Project Plan. The objectives are to: (1) define the effect of processing conditions on the total and free gossypol content in cottonseed products and identify the processing conditions that will produce a product containing consistently low available gossypol to livestock; (2) search for short- and long-term alternative solvents to replace commercial hexane for oilseed extraction; and (3) develop innovative and new processes to enhance the utilization of cottonseed and its co-products. 3. Milestones: A. List the milestones that were scheduled to be addressed in FY2004. How many milestones did you fully or substantially meet in FY2004 and indicate which ones were not fully or substantially met, briefly explain why not, and your plans to do so. Our main objective for FY 2004 was to study the extraction efficiency of a number of alternative hydrocarbon solvents and to determine the compositional properties of the oil and meal products produced with these solvents. In addition, we planned to study the efficiency of a number of oxygenated (oxygen containing) compounds as extraction solvents. Our results indicated that isohexane is a suitable hydrocarbon solvent for the extraction of vegetable oil. This solvent is effective at removing oil from seed material and is not currently considered as a HAP by the Environmental Protection Agency (EPA). Isohexane can be integrated into current oil extraction operations with minimal equipment changes. However, the compound is more expensive than hexane. Our work with oxygenated solvents indicates that acetone may be a suitable solvent. However, moisture is acetone soluble and the influence of moisture on the extraction process needs additional study. B. List the milestones that you expect to address over the next 3 years (FY 2005, 2006, & 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? The current CRIS project will terminate on 8/21/04, and will be replaced by a new project entitled, "New and Expanded Uses of Oilseed Products and By-Products." The objectives for the new project will include (1) developing new industrial uses for cottonseed products and co-products as alternative fuel additives, biobased lubricants, and concrete-form release agents; (2) developing cost-effective methods for producing gossypol from cottonseed and cottonseed by-products; (3) developing new methods and processes for the conversion of oleic and linoleic acids, the most abundant fatty acids of cottonseed, to branched-chain, conjugated, and hydroxylated fatty acids; and (4) developing cost-effective methods for producing new value-added fatty acid-based products from cottonseed processing by developing new process methods for removing gossypol and phospholipids (phosphorous containing lipids) from cottonseed oil miscella (mixture of solvent and oil). Year 1, FY 2005: We will establish protocols for testing oils as fuel additives, lubricants, and concrete-form release agents. As part of this work, we will conduct compositional analysis and preliminary evaluation on cottonseed oil fractions collected from commercial mills and obtained from the laboratory. Optimal conditions will be determined for milling cottonseed to free the pigment glands (containing gossypol) from the other seed tissues. Chemical means of converting oleic and linoleic acids (common fatty acids in vegetable oils) into new branched-chain fatty acids will be found, and gossypol adsorbent experiments will be conducted with cottonseed miscellas. Year 2, FY 2006: Oils with acceptable performance from the FY 2005 study will be tested in factory environments. These oils will be collected from a second crop year and their performance characteristics as fuel additives, lubricants, and concrete form release agents will be re- examined. Experiments will be conducted to identify the desirable and undesirable components in cottonseed oils in the targeted applications. The recovery of gossypol acetic acid (GAA) from isolated cottonseed glands will be studied. Gossypol solubility in acetone will be determined as a function of temperature and the solute crystal form. Studies on the formation of branched-chain fatty acids will be continued, with an emphasis on scale-up and evaluation as biodiesel fuel or alternative fuel additives. Methods will be applied to cottonseed oil and other oils rich in unsaturated fatty acids. Chemical assays for adsorbate (gossypol-rich fraction) and filtrate (gossypol-lean fraction) will be conducted. Adsorbents regeneration will also be studied. A model for adsorption/desorption process will be developed and the process economics will be studied. Phospholipids recovery experiments will also be conducted and the phospholipids will be characterized. Year 3, FY 2007: Experiments will be continued to identify the desirable and undesirable components in cottonseed oils for use as fuel additives, lubricants, and concrete form release agents. We will also conduct compositional analysis and performance evaluation of the various fractions of cottonseed oil to be used in these applications. The separation of organic and aqueous phases during gossypol isolation from cottonseed soapstock has been problematic and will be studied to determine what compositional characteristics lead to phase separation problems. The recovery of chiral gossypol (optically pure isomers of gossypol) will be improved by techniques to enhance simultaneous growth of both optically pure isomeric crystals from the same solution. Studies will continue on oils containing branched-chain fatty acids, with emphasis on scale-up and evaluation of the oils as biodiesel or fuel additives. Studies will also be continued on the conversion of linoleic acid into conjugated unsaturated and hydroxyl fatty acids. Development studies will be conducted on the byproducts from the fatty acids and glycerides (mono-, di- and triglycerides) from supercritical fluid extraction and membrane processes. Products will be characterized. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004 (one per Research Project): Our previous research including bench scale experiments and plant trials has demonstrated that isohexane is an acceptable short term solution in replacing hexane and requires minimum modification for the existing oilseed extraction operation. Long-term alternative to hexane, which is a hazardous air pollutant (HAP), is much more difficult to achieve. Since most of the environmentally safe solvents are generally polar and water-liking, it requires a different set of operation criteria and much more retrofit for the existing extraction facility to use these polar solvents. Our research results indicated that acetone is a viable option to serve as an environmentally safe solvent. Acetone is not volatile organic compound (VOC) nor HAP, yet extracts as effectively as the petroleum paraffins. Acetone does not leach much carbohydrates from the cottonseed flakes as ethanol and isopropanol. The only technical challenge for using acetone appears to be the control of water content in the oilseed material and the reclaimed acetone which is presently being investigated. B. Other Significant Accomplishment(s), if any. About 20 grams of cottonseed oil was produced from cottonseed containing an excess of (-)-gossypol. This sample will be tested for the cancer cell inhibition study by scientists at the Ohio State University. Results will be used to support a research proposal submitted to National Institute of Health (NIH) by the Ohio collaborator. A new processing method for producing a personal care product (Hair styling gel/ingredients) from an oilseed byproduct (soapstock) was developed. An ARS invention disclosure was filed for this product. Commercialization efforts are ongoing. Collaborative research with a scientist at Clemson University demonstrated that monoclonal immunoassay for free gossypol in several cottonseed meals has a good correlation with the official American Oil Chemists' Society (AOCS) Method. The gossypol detection limit for monoclonal assay is about 20 part per billion (ppb) as opposed to 10 parts per million (ppm) for the AOCS method. Approximately 10 grams of (+)- and (-)-gossypol were produced to support a number of internal and external research projects. The compounds were used to support research on the antinutritive effects of gossypol in poultry diets, the insecticidal activity of the compound against the cotton bollworm, the activity of the compounds against cancer cell lines, and the ability of the compound to bind to and inhibit some enzymes. In cooperation with scientists in the Fiber Biosciences Research Group (SRRC), we developed cotton hairy roots from two different species of cotton. Hairy root cultures are composed of root tissues that proliferate independently of other plant organs because they have been infected with a microbe called Agrobacterium rhizogenes. All of the cultures produce gossypol in large amounts (the compound is normally found in cotton plant tissues and seeds). The cultures will be useful for biochemical and genetic analysis of gossypol production and for producing isotope-labeled gossypol for research purposes. C. Significant Accomplishments/Activities that Support Special Target Populations. None. D. Progress Report opportunity to submit additional programmatic information to your Area Office and NPS (optional for all in-house ("D") projects and the projects listed in Appendix A; mandatory for all other subordinate projects). Under a Reimbursable Agreement with Cotton Incorporated (6435-41000-082- 05R), collaborative work was conducted on the specificity of gossypol isomers in the inhibition of cancer cell growth with scientists at the Ohio State University. Research using cottonseed products to control fire ants, termites and other insects is being continued with ARS scientists. The field test with cottonseed product baits showed promising results. We also assisted ARS scientists at Auburn to conduct research to establish the immune or disease-resistance effect of gossypol on catfish. Technical assistance was provided to scientists at the Clemson University in the development of immuno-assay of gossypol in cottonseed products. Under a Specific Cooperative Agreement (6435-41000-082-03S) with Texas A&M Research Center, a sampling plan was developed and communicated to the extrusion mini mills owners. The purpose of the study was to examine the product quality and consistency, gossypol availability in the meal and industrial uses of the oil produced from mills in Texas and South Carolina. Results showed that the oils are darker than usual after refining and bleaching. Finished meal and pellets generally contained higher amounts of free gossypol than meal from the conventional expeller mill. Under a Trust Agreement (6435-41000-082-04T) with Cotton Foundation, a series of hydrogenation tests was carried out for cottonseed oil. When all the linoleic acid (containing 2-double bonds) is converted into monoene fatty acid (with one double bond), the oil becomes significantly more stable than non-hydrogenated oil. From these results, two separate oils were recommended to a concrete roof tile manufacturer to use as a mold-releasing agent. A form of cottonseed oil was suggested to a small company as an ingredient for the dust control application. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Isohexane was developed as an alternative to hexane as an oilseed extraction solvent. The work has encouraged the oilseed extraction industry to replace hexane with isohexane, which is not currently considered a HAP by the EPA. In the last few years, it has been estimated that 20% of U.S. oilseed extraction capacity has converted to isohexane. A gas chromatographic method to measure free fatty acids in oils was developed. The method involves a derivatization of the acids to make them more volatile followed by separation by capillary gas chromatograph. The method correlates well with conventional titration methods. This procedure was adopted by AOCS as a Recommended Practice. Gossypol was proven effective in controlling insects such as, fire ants and termites and a patent application has been approved. Gram quantity of (+)- or (-)-gossypol were produced by a novel crystallization procedure. Many grams of sample have been provided to scientists to support research into the compound's toxicity and potentially useful biological effects. Inclusion complexes of gossypol were formed with several types of ketones. Complexes were made with cyclobutanone, cyclopentanone, cycloheptanone, cyclooctanone, cyclododecanone, 2-pentanone, 3-pentanone, 2-hexanone, 2-heptanone, 3-heptanone, and 4-heptanone were prepared and studied by x-ray diffraction technique. At least four new families of complex have been identified 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Much of our research results have been transferred to the private sectors or professionals in the research communities through technical presentations, annual reports, popular magazine articles and publications in technical journals. Several successful technology transfers during the last few years are described in the following. Many oilseed extraction facilities are motivated by the increasing regulatory pressure and are steadily adopting isohexane as an alternative solvent to replace hexane which is VOC and HAP. This effort will reduce the amount of hazardous air pollutant, n-hexane, to be released by the oilseed extraction operation. The largest cottonseed extraction plant is currently converting from hexane to isohexane. A special refining process was developed in our lab and adopted by Martek Biosciences in the mid 90's. The final product, docosahexaenoic acid (DHA) rich oil which is an omega-3 (with a double bond at the 3rd carbon from the end of a fatty acid) oil found in the mother's milk fat, was approved as a food ingredient by Food and Drug Administration in 2001. The infant formula containing this crucial omega-3 oil became available in the U.S. market in early 2002. Our research effort has demonstrated that Silylation (to make an organic compound more volatile by attaching a silicate functional group at the hydroxyl sites) followed with capillary gas chromatograph can be a good alternative method for the determination of free fatty acid in crude oil or fat. It will not only provide a quantitative information but also a detail distribution of the various fatty acid. This procedure is viewed as a powerful research tool and has been accepted by AOCS as a Recommended Practice. Gossypol, a polyphenolic pigment found in cottonseed and cotton plant tissues, has been known as a natural insecticide. At a low concentration in special tailored bait system, the gossypol can be an attractant to termite. Eventually, the entire termite colony will be weakened and destroyed by the biological agent. A patent on the termite control with gossypol containing bait system was approved by the U.S. Patent Office. A Japanese company is currently evaluating the technology. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. 1. "Natural Style," Soap, Perfumery and Cosmetics, 76:53, UK, 2003. 2. "Veggie Oil-based Hair Care Products," Agricultural Research, 51:19, 2003

Impacts
(N/A)

Publications

  • Yildirim-Aksoy, M., Lim, C.E., Dowd, M., Wan, P., Klesius, P.H., Shoemaker, C.A. 2004. In vitro inhibitory effect of gossypol from gossypol-acetic acid, and (+)- and (-)-isomers of gossypol on the growth of Edwardsiella ictaluri. Journal of Applied Microbiology. 97:97-82.
  • Moss, S.C., Triplett, B.A., Dowd, M.K. 2004. Initiation and proliferation of cotton hairy roots [abstract]. Congress for In-Vitro Biology. p. 53A.
  • Dowd, M.K., Stevens, E.D. 2004. The inclusion complex of (-)-gossypol with 2,4-pentanedione (abstract}. American Oil Chemists' Society. p.123
  • Wan, P.J., Waggoner, A.E., Dowd, M.K. 2004. Effect of extraction solvents on composition of oil and meal obtained from cottonseed (abstract). American Oil Chemists' Society. p. 123-124.
  • Kuk, M.S., Bland, J.M. 2004. Molecular species of cottonseed phospholipids and gossypol from cottonseed processing (abstract). American Oil Chemists' Society. p. 103.
  • Wakelyn, P.J., Wan, P.J. 2004. Edible oil extraction solvents: fda regulatory considerations. Inform. 15(1):22-23


Progress 10/01/02 to 09/30/03

Outputs
1. What major problem or issue is being resolved and how are you resolving it? Product quality, processing efficiency and by-product utilization of oilseeds continue to be the most challenging issues of the oilseed industry and the major areas of our research focus. Both internal and outside resources are applied to the problems solving effort. Outside resources include oilseed industry, professional associations such as, National Cottonseed Products Association (NCPA), Cotton Inc., Cotton Foundation, etc., and university professors. More specifically, a lot of effort was placed in improving the safe and value-added uses of cottonseed products and its by-products. To accomplish this, cottonseed products are produced under defined conditions and these products are analyzed for total and free gossypol(natural pigment present in cottonseed products) content as described in the Methods of the American Oil Chemists' Society. At the same time, these products are fed to selected livestock by scientists at the Texas AM University to determine the gossypol availability in the processed cottonseed products to ruminant (multiple stomach) and non-ruminant (single stomach) animals. Other process improvements and value added uses of cottonseed products and its by-products are being studied in the ARS lab and pilot plant. 2. How serious is the problem? Why does it matter? Oilseed industry in the United States experiences increasing competition and regulatory pressure. Therefore, there is always an urgent need to improve the productivity of their processes and product quality or to enhance by-product utilization. The profit margin of this industry is relatively low and further depressed in recent years due to worldwide over supply. It is especially true for the cottonseed processors who can not afford to have any in-house research activity. Collectively, through NCPA, Cotton Inc. and Cotton Foundation, the cottonseed processors have expressed their technical needs and challenges to us which in turn became our research priorities. One example, major changes in the commercial processing of cottonseed through the years have resulted in questions regarding the safe use of cottonseed products as a feed ingredient. Increasing the feed safety of the resulting meals is expected to expand the use of cottonseed meal in non-ruminant feeds. Another example, due to increasing regulatory pressure, an alternative and safer solvent than hexane, its main component has been classified as one of the 189 Hazardous Air Pollutants by US Environmental Protection Agency (EPA), has always been an urgent long-term objective for the oilseed extraction industry. New value-added uses of oilseed co-products should also increase the marketability and economical viability of the corresponding crop. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? The research effort is to improve the quality of cottonseed products through process innovation and to enhance their marketability via value- added new uses of the various co-products of cottonseed and other oilseeds. These CRIS goals relate to National Program 306: New Uses, Quality, and Marketability of Plant Products. 4. What were the most significant accomplishments this past year? A better understanding and value-added use of co-product of cottonseed has always been regarded as a means to bring improved marketability to the industry. Gram quantities of optically pure gossypol were produced in the ARS lab and provided to scientists at the University of Georgia. Results indicated that (-) gossypol has much greater impact to the reduction of growth performance in chicken than (+) gossypol. These findings should lead to a clearer guidance in the feeding practice for the poultry industry who are using cottonseed meal as a protein ingredient. B. Other Significant Accomplishment(s), if any. (1) The industrial interest in using isohexane (isomers of hexane) to replace hexane in order to reduce the Environmental Protection Agency regulatory pressure has gained further momentum. A short technical session on this subject was organized at the annual convention of American Oil Chemists' Society in Kansas City, MO, May 5-7, 2003, with the speakers from ARS, supplier and users of commercial isohexane. This is the first time to have the commercial users of isohexane share their experience. The users acknowledged that there is minor energy savings by changing from hexane to isohexane. The greatest advantages of using isohexane appear to be in the reduction of environmental pressure and lowered emission loss of solvent. Cargill, being one of the largest oilseed processors in the world, indicated that they have converted all their extraction facilities to isohexane which include soy, corn and sunflower. (2) Refining cottonseed oil to a consistently light color has been a challenging issue for the industry. A unique cottonseed oil refining technique was developed in the ARS lab and the same refining technique has assisted a bioscience firm to successfully commercialize docosahexaenoic acid (DHA) rich oil obtained from algae. This same DHA oil, along with arachidonic acid (AA), has been approved by the Food and Drug Administration in 2001 to be added to the infant formula. Worldwide more than 60 nations already used DHA and AA in their baby formula. The first commercially available infant formula containing DHA and AA with a trade name of Lipil was made available to consumers in the U.S. in the spring of 2002 by Mead Johnson. Eighty cans of Lipil were supplied as one of the products for the Congressional Budget Hearing for the summer of 2003. (3) ARS scientists at Southern Regional Research Center continued to work collaboratively with (a) Food Protein RD Center, Texas AM University to determine the feasibility of using acetone, a more polar solvent than hexane, as an alternative solvent to extract oilseed and (b) an ARS colleague at the Auburn University to determine the potential benefit of immune or disease-resistant improvement by gossypol in cottonseed meal which was used in the catfish feed. (4) Further modifications of the patent application document on cottonseed products used for termite control were suggested by the patent examiner and revised documents were submitted to the U.S. Trade Mark and Patent Office. A Japanese company has expressed an interest to license the technology. (5) Large-scale application tests for utilizing the biodegradable films produced from oilseed byproducts (cottonseed, soybean and safflower) was conducted. The statistical analyses indicated that the oilseed-derived- film can extend the postharvest shelf life of freshly harvested cucumbers and bell peppers. (6) About 100 grams of cottonseed oils from three different varieties of cottonseed containing various ratio of (-) gossypol and (+) gossypol were produced in the ARS lab. These samples were tested for the cancer cell inhibition study by scientists at the Ohio State University. Results indicated that oil containing higher amount of (-) gossypol have much better inhibition effect to cancer cell growth. Three different research proposals were submitted by the collaborator during 2003. (7) A new processing method for producing a personal care product (Hair styling gel/ingredients) from an oilseed byproduct (soapstock) was developed. An ARS invention disclosure for this new products/ingredients was filed. Commercialization efforts are underway. (8) Inclusion complexes of gossypol with several types of ketone were prepared. Complexes with cyclobutanone, cyclopentanone, cycloheptanone, cyclooctanone, cyclododecanone, 2-pentanone, 3-pentanone, 2-hexanone, 2- heptanone, 3-heptanone, and 4-heptanone were prepared and studied by x- ray diffraction technique. At least two new families of complex have been identified. (9) Cotton Foundation provided a research grant of $12,000 (covering July 1, 2002 to June 30, 2003) and Cotton Inc. provided a research grant of $25,000 (January 1 -December 31, 2003) to support ARS efforts in searching for industrial and value-added uses of cottonseed products. C. Significant Accomplishments/Activities that Support Special Target Populations. None. D. Progress Report (OPTIONAL) (1) This part of the progress report is done under a Reimburseable Agreement with Cotton Incorporated. Collaborative work on the specificity of gossypol isomers in the inhibition of cancer cell growth is being continued with the scientists at the Ohio State University. Research using cottonseed products to control fire ants, termites and other insects is being continued with ARS scientists. The field test with the cottonseed products containing baits for termites in the Mississippi region showed promising result. The second round analysis confirmed the original observation that cottonseed products indeed contain high level of melatonin which is a hormone essential to sleep pattern. This work is done in cooperation with scientists at the Health Science Center of the University of Texas in San Antonio, Texas. We also assisted ARS scientists at Auburn to conduct research to establish the toxicity level (when the growth performance was affected) of gossypol in the feed to catfish and to study the immune or disease-resistance effect of gossyol on catfish. Technical advices and assistance were provided to scientists at the Clemson University in the development of immuno-assay of gossypol in cottonseed products. (2) This part of the progress report is done under a Reimburseable Agreement with Texas AM Research Center at San Angelo, Texas. A sampling plan was developed and communicated to the extrusion mini mills owners. The purpose of the study was to examine the product quality and consistency, gossypol availability in the meal and industrial uses of the oil produced from these mills. Seven different mini mills were visited. Duplicate samples were collected from each of these mills. Results showed that the oils are darker than usual after refining and bleaching. Finished meal and pellets generally contained higher amounts of free gossypol, which is considered more toxic and is measured by the method of American Oil Chemists' Society (AOCS), than meal from the conventional expeller mill. One third of the book chapters was collected for the monograph on "Gossypol and Cottonseed - Chemistry, Processing and Utilization" which will be published by AOCS Press in the spring of 2004. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. (1) Oil Color: (a) An international collaborative study organized by ARS scientists led to the establishment of an official method published by AOCS in 1998. (b) A refining process condition was developed to deal with dark oils and has been used successfully by a specialty oil producer. (2) Isohexane as an Alternative to Hexane: Isohexane was demonstrated to be a cost efficient solvent. In addition, it is not toxic to nerve systems. The work at the Center led to the publication of a monograph "Technology and Solvents for Extracting Oilseeds and Nonpetroleum Oils" by AOCS Press in 1997, and it encouraged the oilseed extraction industry to replace hexane with isohexane. (3) Gossypol Analysis: High Performance Liquid Chromatograph method was developed to determine gossypol and its optical isomers. It is quick and environmentally friendly. (4) Free Fatty Acid (FFA) Analysis: The existing official method of AOCS (Aa 6-38) has been proven to underestimate the amount of FFA in cottonseed by as much as 10%. Recommendation of improved approach for the determination of FFA in cottonseed was approved and adopted by the Method Committee of AOCS. (5) Knowledge about the biological availability gossypol of cottonseed products to livestock has been generated and periodically presented to the public through professional meetings during the last few years. Some of the findings are summarized in the following. Processing condition can drastically affect the amount of free gossypol, which is the more toxic form to livestock, present in the final products. Screw pressed meal generally contains lower free gossypol due to high shear pressure and temperature and therefore low gossypol availability to lamb than expander-solvent extracted meals. Ferrous sulfate can effectively reduce the gossypol availability of cottonseed products and soapstock to lamb. Gossypol in soapstock is not readily available to lamb. High free fatty acid in cottonseed did not increase the gossypol availability to lamb. (6) Gossypol was proven effective in controlling insects such as, fire ants and termites and a patent application has been filed. (7) Gossypol in cottonseed oil can inhibit cancer cell growth. The left-handed isomer of gossypol, (-) gossypol, was demonstrated to have the greatest effect on cancer cells than (+)-gossypol. 6. What do you expect to accomplish, year by year, over the next 3 years? FY2004: (1) Define process conditions to produce consistently low free gossypol cottonseed meal. (2) Complete the collaborative study on gossypol analysis by HPLC. (3) Explore value-added uses of cottonseed products and its derivatives. (4) Complete the monograph on "Gossypol and Cottonseed - Chemistry, Processing and Utilization." (5) Improve analytical procedure for gossypol in refined-bleached- deodorized oil. (6) Evaluate product quality from extrusion mills for the second year. (7) Define the safety level of gossypol in the diet of catfish through a collaborative effort with an ARS scientist in Auburn, AL. (8) Study the processing effect on melatonin in cottonseed and its co- products under a cooperative study with Professor Reiter at the Health Science Center, Univ. of Texas at San Antonio, TX. FY2005: Investigate innovative processing conditions and formulation to reduce gossypol availability in cottonseed products. Value-added uses of oilseed products and components. Study the most effective form and concentration of gossypol in nutraceutical applications and insect remediations. FY2006: Investigate the optimum dosage of gossypol and gossypol containing products in nutraceutical applications and insect control applications. Elucidate the mechanism of gossypol reactions in selected nutraceutical applications and insect control applications. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? (1) A quick, simple test for free gossypol method has been adopted by the AOCS to be used for screening of gossypol content by cottonseed breeders. (2) Isohexane as an alternative solvent to replace hexane has been slowly accepted by the oilseed crushing industry. It will reduce the amount of hazardous air pollutant, n-hexane, to be released by the oilseed extraction operation. (3) A special refining process was adopted by Martek Biosciences. The final product, DHA rich oil, was approved by FDA in 2001 and is now added into the infant formula. (4) Automated oil color measurement has been adopted by AOCS as an official method. (5) The existing FFA determination for cottonseed which has been used for nearly a century will be discarded and the Method Committee of AOCS has approved and adopted the recommendations based on our recent collaborative study with labs from the cottonseed industry. The same modified method will be included in the Trading Rules of NCPA. (6) A quick and simple hydraulic press method was developed for the estimation of FFA in white cottonseed. It has been routinely used by a cottonseed mill in Georgia. (7) Once-refined cottonseed oil has been used in the formulation of concrete roof- tile mold-release agent. Further study is underway to minimize the polymerization problem of cottonseed oil. (8) A patent on the termite control with gossypol containing bait system is pending. A Japanese company expressed interest to license the technology. 8. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: This does not replace your peer-reviewed publications listed below). Wan, P.J. A Link in the Chain from Oil Refinement to Baby Formula. Agricultural Research. 2002. v.50(12):10-11. Kuk, M.S. Scientist develops Eco-Friendly and Inexpensive Hair Gel From Soybean and Safflower Seeds. ARS News Information, ARS, USDA, 02/25/2003. Kuk, M.S. Oilseed Byproduct Yield Low-cost Lipids for Hair-Gel. Chemical Engineering. 2003. v.110(3):15-21.

Impacts
(N/A)

Publications

  • Wakelyn, P. J., Wan, P.J. Extraction solvents: safety, health and environmental issues. Tzia, C., Liadakis, G., editors. Marcel Dekker, Inc. New York, NY. Extraction Optimization in Food Engineering. 2003. p. 391- 427.
  • Wan, P.J., Zarins, Z. Specific heat of cottonseed and its co-products. Journal of American Oil Chemists' Society. 2003. v. 80(2) p. 123-126.
  • Yildirim, M., Lim, C., Wan, P., Klesius, P.H. Growth performance and immune response of channel catfish fed diets containing graded levels of gossypol acetate. Journal of World Aquaculture Society. 2003. v. 34(1). p. 1-18.
  • Dowd, M.K. Preparation of enantiomeric gossypol by crystallization. Chirality. 2003. v. 15. p. 486-493.
  • Dowd, M.K., Stevens, E.D. The gossypol-cyclododecanone (1/2) inclusion complex. Acta Crystallography. Section C. 2003. v. 59. p. 397-399.
  • Dao, V.-T., Dowd, M.K., Gaspard, C., Martin, M.-T., Hemez, J., Laprevote, O., Mayer, M., Michelot, R.J. New thioderivatives of gossypol and gossypolone, as prodrugs of cytotoxic agents. Bioorganic Medical Chemistry. 2003. v. 11. p. 001-2006.
  • Kuk, M.S. Cottonseed phospholipid. Book of Abstracts. 94th American Oil Chemists' Society Annual Meeting. Kansas City, MO. 2003. Abstract p. 103.
  • Dowd, M.K. Stevens, E.D. Gossypol inclusion complexes with 2-pentanone, 3- pentanone, and 2-hexanone. Book of Abstracts. 94th American Oil Chemists' Society Annual Meeting. Kansas City, MO. 2003. Abstract p. 117.


Progress 10/01/01 to 09/30/02

Outputs
1. What major problem or issue is being resolved and how are you resolving it? Product quality, processing efficiency and by-product utilization of oilseeds continue to be the most challenging issues of the oilseed industry and the major areas of our research focus. Both internal and outside resources are applied to the problems solving effort. Outside resources include oilseed industry, professional associations such as, National Cottonseed Products Association (NCPA), Cotton Inc., Cotton Foundation, etc., and university professors. More specifically, a lot of effort was placed in improving the safe and value-added uses of cottonseed products and its by-products. To accomplish this, cottonseed products are produced under defined conditions and these products are analyzed for total and free gossypol. At the same time, these products are fed to selected livestock by scientists at the Texas A&M University to determine the gossypol availability in the processed cottonseed products to ruminant and non-ruminant animals. Other process improvements and value added uses of cottonseed products and its by- products are being studied in the lab and pilot plant. 2. How serious is the problem? Why does it matter? The Oilseed industry in the United States experiences increasing competition and regulatory pressure. Therefore, there is always an urgent need to improve the productivity of their processes and product quality or to enhance by-product utilization. The profit margin of this industry is generally on the low side and extremely depressed in recent years. It is especially true for the cottonseed processors who can not afford to have any in-house research activity. Collectively, through NCPA, Cotton Inc. and Cotton Foundation, the cottonseed processors have expressed their technical needs and challenges to us which in turn became our research priorities. One example, major changes in the commercial processing cottonseed through the years have resulted in questions regarding the safe use of cottonseed products as a feed ingredient. Increasing the feed safety of the resulting meals is expected to expand the use of cottonseed meal in non-ruminant feeds. Another example, due to increasing regulatory pressure, an alternative and safer solvent than hexane has always been an urgent long term objective for the oilseed extraction industry. New value-added uses of oilseed co-products should also increase the marketability and economical viability of the corresponding crop. 3. How does it relate to the national Program(s) and National Program Component(s) to which it has been assigned? The research effort is to improve the quality of cottonseed products through process innovation and to enhance their marketability via value- added new uses of the various co-products of cottonseed and other oilseeds. These CRIS goals relate to National Program 306: New Uses, Quality, and Marketability of Plant Products. 4. What was your most significant accomplishment this past year? A. Single Most Significant Accomplishment during FY 2001 year: The Environmental Protection Agency (EPA) already published a Maximum Achievable Control Technology (MACT) standards for oilseed extraction industry early 2001. The oilseed industry is expected to comply with the hexane loss standards by 2005. This new regulation further prompts the industry to consider an alternative solvent to replace the commercial hexane used in their facility. Our research results suggested that commercial isohexane can be a good alternative solvent for hexane and we collaborated with two different cottonseed oil mills to test the commercial isohexane. The plant trials proved that isohexane can be used effectively and offers the opportunity to save energy, increase productivity and reduce the emission of n-hexane. These research results have encouraged the oilseed extraction industry willingly to examine commercial isohexane in their oil mills or to convert to isohexane from hexane. A private company has requested our assistance to convert one of their soybean extraction facilities from hexane to isohexane in April, 2001. The results showed some energy savings however not as much as those reported by the smaller cottonseed processing facilities. But the trials with isohexane at an old soy extraction plant were encouraging and the experience will be incorporated in their design and construction of a new soy extraction facility. B. Other Significant Accomplishments(s), if any. (1) A unique cottonseed oil refining technique has assisted a bioscience firm to successfully commercialize docosahexaenoic acid (DHA) rich algal oil. This same DHA oil along with arachidonic acid (AA) have been approved by FDA in 2001 to be added to the infant formula. Worldwide more than 60 nations already used DHA and AA in their baby formula. (2) The Method Committee of American Oil Chemists' Society (AOCS) has adopted the silylation and capillary GC procedure for the determination of free fatty acid (FFA) and FFA profile in crude vegetable oils as a Recommended Practice. (3) Worked collaboratively with Food Protein R&D Center, Texas A&M University to determine the feasibility of using acetone as an alternative solvent to extract oilseed. (4) Continued collaborative research with an ARS colleague at Auburn University to determine the toxicity level of gossypol when cottonseed meal was used in the cat fish feed. (5) Application of cottonseed products for termite control has been successful and one patent application was submitted to the Trade Mark and Patent Office. (6) Gram quantity of pure isomeric gossypol [8 g of (-)-gossypol and 8 g of (+)-gossypol by crystallization] have been produced and supplied to other labs for various biological studies. 3 g of each have been sent to Stipanovic, 1 g of each to Michelot, 100 mg of each to Mellon. (7) Cotton Foundation provided a research grant of $10,000 (covering from July 1, 2001 to June 30, 2002) to support our effort in searching for industrial and value-added uses of cottonseed products. Our technical support has lead to the use of cottonseed oil in the concrete roof-tile mold-releasing agent. When fully implemented, a single company would demand 7,000 lbs of cottonseed oil. In a separate effort, it was demonstrated that the detection limit of gossypol in refined-bleached- deodorized cottonseed oil was 0.5 ppm with the best known available procedure. (8) Corrected the structure of gossypol-acetone (1:3), which we initially reported as gossypol-acetic acid (1:3). Diffraction work at lower temperatures better resolved the acetone molecules. (9) Studied gossypol inclusion phenomena with a number of solvents. Purpose was to try to find better preparative methods. Identified several new gossypol inclusion complexes. Structures solved by diffraction for gossypol-MEK (1:1), gossypol-MEK (1:2), (-)-gossypol-MEK (1:3), gossypol-3-pentanone (2:1), gossypol-2-pentanone (2:1), gossypol-2- hexanone (2:1). (10) A variety of Sea Island barbadense cotton was propagated in the greenhouse over the winter. Purpose was to produce sufficient seed to plant a field. The field crop is planted. This material will be used to try to isolated methyl gossypol derivatives. (11) A chemical reaction condition which governs the conversion of gossypol from the free to the bound form was investigated. This finding could be applied to the cottonseed processing. (12) Cottonseed soapstock is one of the most underutilized by-products from oilseed processing. Applying a few chemical and physical treatments to various soapstocks obtained from oilseed processing, SRRC scientists demonstrated that cottonseed, safflower and soybean soapstocks can be utilized to make new products for extending shelf-life of fresh produce. This progress report is done under a Reimburseable Agreement with Cotton Incorporated. Collaborative work on the specificity of isomeric gossypol in the inhibition of cancer cell growth is being continued with the scientists at the Ohio State University. Research using cottonseed products to control fire ants, termites and other insects is being continued. The field test with the cottonseed products containing baits for termites in the Mississippi region showed promising results. The second round analysis confirmed the original observation that cottonseed products indeed contain high level of melatonin. We assisted ARS scientists at Auburn to conduct research to establish the toxicity level of gossypol in the feed to catfish and to study the immune effect of gossyol on catfish. This progress report is done under a Reimburseable Agreement with Texas A&M Research Center at San Angelo, Texas. A sampling plan was developed and communicated to the extrusion mini mills owners. The purpose of the study is to examine the product quality and consistency, gossypol availability in the meal and industrial uses of the oil produced from these mills. Seven different mini mills were visited. Duplicate samples were collected from each of these mills. Results showed that the oils are darker than usual after refining and bleaching. Finished meal and pellets generally contained higher amounts of free gossypol than meal from conventional expeller mill. The gossypol availability of these meals and pellets in lamb feeding is being investigated. C. Significant Accomplishments/Activities that Support Special Target Populations. None 5. Describe your major accomplishments over the life of the project, including their predicted or actual impact? (1) Oil Color: (a) An international collaborative study organized by us and led to the establishment of an official method published by AOCS in 1998. (b) A refining process condition was developed to deal with dark oils which has been used successfully by a specialty oil producer. (2) Isohexane as an Alternative to Hexane: Isohexane was demonstrated to be a cost efficient solvent besides it is not a neural toxicant. The work at the Center led to the publication of a monograph "Technology and Solvents for Extracting Oilseeds and Nonpetroleum Oils" by AOCS Press in 1997 and encouraged the oilseed extraction industry to replace hexane with isohexane. (3) Gossypol Analysis: HPLC method was developed to determine gossypol and its isomers. It is quick and environmentally friendly. (4) Free Fatty Acid Analysis: The existing official method of AOCS (Aa 6-38) has been proven to underestimate the amount of FFA in cottonseed by as much as 10%. Recommendation of improved approach for the determination of FFA in cottonseed was approved and adopted by the Method Committee of AOCS. (5) Knowledge about the gossypol availability in cottonseed products to livestock has been generated during the last few years. Screw pressed meal contains on an average of lower free gossypol and therefore low gossypol availability to lamb than expander-solvent extracted meals. Ferrous sulfate can effectively reduce the gossypol availability of cottonseed products and soapstock to lamb. Gossypol in soapstock is not readily available to lamb. High free fatty acid in cottonseed did not increase the gossypol availability to lamb. (6) Gossypol was proven effective in controlling insects such as, fire ants and termites and a patent application has been filed. 6. What do you expect to accomplish, year by year, over the next 3 years? FY2003: Define process conditions to produce consistently low free gossypol cottonseed meal. Complete the collaborative study on gossypol analysis by HPLC. Explore value-added uses of cottonseed products and its derivatives. Complete the monograph on "Gossypol and Cottonseed - Chemistry, Processing and Utilization". Improve analytical procedure for gossypol in refined-bleached-deodorized oil. Evaluate product quality from extrusion mills for the second year. Define the safety level of gossypol in the diet of catfish through a collaborative effort with an ARS scientist in Auburn, AL. Study the processing effect on melatonin in cottonseed and its co- products under a cooperative study with Professor Reiter at the Health Science Center, Univ. of Texas at San Antonio, TX. FY2004: Investigate innovative processing conditions and formulation to reduce gossypol availability in cottonseed products. Value-added uses of oilseed products and components. Study the most effective form and concentration of gossypol in nutraceutical applications and insect remediations. FY2005: Investigate the optimum dosage of gossypol and gossypol containing products in nutraceutical applications and insect control applications. Elucidate the mechanism of gossypol reactions in selected nutraceutical applications and insect control applications. 7. What technologies have been transferred and to whom? When is the technology likely to become available to the end user (industry, farmer other scientist)? What are the constraints, if known, to the adoption durability of the technology? Quick, simple test for free gossypol method has been adopted by the AOCS to be used for screening of gossypol content by cottonseed breeders. Isohexane as an alternative solvent to replace hexane which has been slowly accepted by the oilseed crushing industry. A special refining process was adopted by Martek Biosciences. Automated oil color measurement has been adopted by AOCS as an official method. Existing FFA determination for cottonseed which has been used for nearly a century will be discarded and the Method Committee of AOCS has approved and adopted the recommendations based on our recent collaborative study. The same modified method will be included in the Trading Rules of NCPA. A quick and simple hydraulic press method was developed for the estimation of FFA in white cottonseed. It has been routinely used by a cottonseed mill in Georgia.

Impacts
(N/A)

Publications

  • 1. Wan, P.J., Dowd, M.K. American Oil Chemists' Society Recommended Practice Ca 5d-01 "Free fatty acids in crude vegetable oils by capillary gas chromatograph". The 7th Edition of the Official Methods and Recommended Practices of the AOCS. Campaign, IL. 2001.
  • 2. Wakelyn, P.J., Hron, R.J., Flider, F.J., Wan, P.J. Acetone: An Environmentally Preferable Choice for Oilseed Extraction? INFORM. 200l. v. 12. p. 887-893.
  • 3. Dowd, M.K., Thomas, L.M., Calhoun, M.C. Correction to the structure of enantiomeric gossypol. Journal of American Oil Chemists' Society. 2001. v. 78. p. 1171.
  • 4. Wan, P. J., Zarins, Z.M. Specific heats of cottonseed and its co- products. A Special Supplement to INFORM. 2002. v. 12. p. 154.


Progress 10/01/00 to 09/30/01

Outputs
1. What major problem or issue is being resolved and how are you resolving it? Product quality, processing efficiency and by-product utilization of oilseeds continue to be the most challenging issues of the oilseed industry and the major areas of our research focus. Both internal and outside resources are applied to the problem solving effort. Outside resources include the oilseed industry, professional associations such as, National Cottonseed Products Association (NCPA), Cotton Inc., Cotton Foundation, etc., and University professors. More specifically, a lot of effort was placed in improving the safe and value-added uses of cottonseed products and its by-products. To accomplish this, cottonseed products are produced under defined conditions and then analyzed for total and free gossypol. At the same time, these products are fed to selected livestock by scientists at the Texas A&M University to determine the gossypol availability in the processed cottonseed products to ruminant and non-ruminant animals. Other process improvements and value-added uses of cottonseed products and its by-products are being studied in the lab and pilot plant. 2. How serious is the problem? Why does it matter? The oilseed industry in the United States experiences increasing competition and regulatory pressure. Therefore, there is always an urgent need to improve their processes and product quality and/or to enhance by-product utilization. The profit margin of this industry is generally on the low side and has been extremely depressed in recent years. It is especially true for the cottonseed processors who can not afford to have any in-house research activity. Collectively, through NCPA, Cotton Inc. and Cotton Foundation, the cottonseed processors have expressed their technical needs and challenges to Agricultural Research Service (ARS) scientists at the Southern Regional Research Center (SRRC) which in turn became our research priorities. One example, major changes in the commercial processing of cottonseed through the years has resulted in questions regarding the safe use of cottonseed products as a feed ingredient. Increasing the safety of the resulting meals is expected to expand the use of cottonseed meal in non-ruminant feeds. Another example, due to increasing regulatory pressure, an alternative and safer solvent than hexane has always been an urgent long-term objective for the oilseed extraction industry. New value-added uses of oilseed co-products should also increase the marketability and economical viability of the corresponding crop. 3. How does it relate to the National Program(s) and National Component(s)? The research effort is to improve the quality of cottonseed products through process innovation and to enhance their marketability via value-added new uses of the various co-products of cottonseed and other oilseeds. This work is a component of National Program 306: Quality and Utilization of Agricultural Products [New Processes, New Uses, and Value-Added Biobased Products] (100%). 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2001 year: The Environmental Protection Agency (EPA) has already published a Maximum Achievable Control Technology (MACT) standard for the oilseed extraction industry in early 2001. The oilseed industry is expected to comply with the amount of commercial volatile hydrocarbons (hexane) loss standards (amount that can be released into the atmosphere) by 2005. This new regulation further prompts the industry to consider an alternative solvent to replace the hexane used in their facility. Our research results suggested that commercial isomers of hexane (isohexane) can be a good alternative solvent for hexane and we collaborated with two different cottonseed oil mills to test the commercial isohexane. The plant trials proved that isohexane can be used effectively and offers the opportunity to save energy and increase productivity. These research results have encouraged the oilseed extraction industry to willingly examine commercial isohexane in their oil mills or to convert to isohexane from hexane. A private company has requested our assistance to convert one of their soybean extraction facility from hexane to isohexane in April, 2001. So far the results are encouraging. B. Other Significant Accomplishment(s), if any. (1) Free fatty acid (FFA) in cottonseed: (a) The report and recommendation based on the findings of a collaborative study were accepted and adopted by the Method Committee of American Oil Chemists' Society (AOCS). The same recommendation has also been adopted by the NCPA Trading Rules. This effort will minimize the erroneous determination of cottonseed grade and provide a more accurate assessment of the refining loss of cottonseed oil. (b) Quick FFA method using a hydraulic press has been proven reproducible and ready to be demonstrated to the cottonseed industry and in the field. (c) Silylation(making volatile derivatives of FFA with a silicon compound) and capillary Gas Chromatography (GC) to determine FFA and FFA profile in crude oils has been established. A Recommended Practice has been proposed to the AOCS Method Committee. (2) Extraction efficiency of pure components of hexane and isohexane and other common polar solvents such as, acetone, ethanol, isopropanol, has been evaluated with a lab scale extractor. (3) Both glandless and glanded cottonseed meals were prepared in the pilot plant of Texas A&M University under a cooperative research agreement and are being fed to catfish by an ARS colleague at Auburn University to determine the toxicity level for cottonseed meal. (4) Ideas of by-products utilization have been assessed including isolation and purification of gossypol from soapstock. Research in value-added uses of meal, oil, gossypol and soapstock are being explored at the SRRC and in cooperation with several university professors. Application of cottonseed products for termite control has been successful and resulted in one patent application. (5) Separation of pure (+) and (-) isomers of gossypol by a crystallization process has been accomplished. Milligram to gram quantities of pure isomeric gossypol have been produced and used for medicinal and biological research. A (-) gossypol was proven much more active in cancer cell inhibition than (+) gossypol in a model system by a professor in the Ohio State University. (6) Cotton Foundation provided a research grant of $10,000 to support our effort in searching for industrial and value-added uses of cottonseed products. The funding period covers from July 1, 2001 to June 30, 2002. C. Significant Accomplishments/Activities that Support Special Target Populations. None. D. Progress Report. (1) This part of the progress report is done under a Reimbursable Agreement with Cotton Incorporated. A (-) isomer of gossypol proven to be much more potent than (+) isomer of gossypol in the inhibition of cancer cell growth under a cooperative research effort between ARS and scientists at Ohio State University. Other value-added uses of cottonseed and its derivatives led to the use of cottonseed products to control fire ants and termites. A patent has been drafted and submitted. A field test with the cottonseed products containing baits has been initiated in Mississippi. Preliminary results were encouraging. Since the preliminary analysis of cottonseed products showed that it is uniquely high in melatonin (a hormone produced by the pineal gland), a second round analysis has been initiated to confirm the original observation. This work is done through a cooperative effort with scientists at the Health Science Center of the University of Texas in San Antonio, Texas. Assisted the scientists at ARS Auburn station in conducting research to establish the toxicity level of gossypol in the feed to catfish and to study the immune effect of gossypol on catfish. (2) Under a Specific Cooperative Agreement with Texas A&M Research Center at San Angelo, Texas, a sampling plan was developed and communicated to the extrusion mini mills owners. The purpose of the study is to examine the product quality and consistency, gossypol availability in the meal and industrial uses of the oil produced from these mills. A collaborative study using High Performance Liquid Chromatography (HPLC) to determine gossypol content and (+) and (-) isomeric gossypol composition was initiated. Six labs participated in the first round effort. Further collaborative tests are needed in order to establish the HPLC method as an accepted official method by AOCS. The effort to collect and write book chapters was continued for the monograph on "Gossypol and Cottonseed - Chemistry, Processing and Utilization" which will be published by AOCS Press. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. (1) Oil Color: (a) An international collaborative study organized by ARS scientists at SRRC led to the establishment of an official method published by AOCS in 1998. (b) A refining process condition was developed to deal with dark oils which has been used successfully by a specialty oil producer. (2) Isohexane as an Alternative to Hexane: Isohexane was demonstrated to be a cost-efficient solvent besides it is not a neural toxicant. The work at the Center led to the publication of a monograph "Technology and Solvents for Extracting Oilseeds and Nonpetroleum Oils" by AOCS Press in 1997 and encouraged the oilseed extraction industry to replace hexane with isohexane. (3) Gossypol Analysis: HPLC method was developed to determine gossypol and its isomers. It is quick and environmentally friendly. (4) Free Fatty Acid (FFA) Analysis: The existing official method of AOCS (Aa 6-38) has been proven to underestimate the amount of FFA in cottonseed by as much as 10%. Recommendation of improved approach for the determination of FFA in cottonseed was approved and adopted by the Method Committee of AOCS. (5) Knowledge about the gossypol availability in cottonseed products to livestock has been generated during the last few years. Screw-pressed meal contains on an average lower free gossypol and, therefore, lower gossypol availability to lambs than expander-solvent extracted meals. Iron salts (Ferrous sulfate) can effectively reduce the gossypol availability of cottonseed products and soapstock to lamb. Gossypol in soapstock is not readily available to lamb. High free fatty acid in cottonseed did not increase the gossypol availability to lamb. (6) Gossypol was proven effective in controlling insects, such as fire ants and termites, and a patent application has been filed. 6. What do you expect to accomplish, year by year, over the next 3 years? FY 2002 - Define process conditions to produce consistently low free gossypol cottonseed meal. Complete the collaborative study on gossypol analysis by HPLC. Explore value-added uses of cottonseed products and its derivatives. Complete the monograph on "Gossypol and Cottonseed - Chemistry, Processing and Utilization". Explore a confirmational analytical procedure for gossypol in refined-bleached-deodorized oil. Evaluate product quality from extrusion mills. Define the safety level of gossypol in the diet of catfish through a collaborative effort with an ARS scientist in Auburn, AL. Confirm the presence of melatonin in cottonseed and its co-products under a cooperative study with Professor Reiter at the Health Science Center, University of Texas at San Antonio, TX. FY 2003 - Investigate innovative processing conditions and formulation to reduce gossypol availability in cottonseed products. Study the most effective form and concentration of gossypol in nutritional and medicinal (nutraceutical) applications and insect remediations. FY 2004 - Investigate the optimum dosage of gossypol and gossypol containing products in nutraceutical applications and insect control applications. Elucidate the mechanism of gossypol reactions in selected nutraceutical applications and insect control applications. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end user (industry, farmer, other scientists)? What are the constraints if known, to the adoption & durability of the technology product? (1) Quick, simple test for free gossypol method has been adopted by the AOCS to be used for screening of gossypol content by cottonseed breeders. (2) Isohexane as an alternative solvent to replace hexane which has been slowly accepted by the oilseed crushing industry. (3) A special refining process was adopted by Martek Biosciences. (4) Automated oil color measurement has been adopted by AOCS as an official method. (5) Existing FFA determination for cottonseed which has been used for nearly a century will be discarded and the Method Committee of AOCS has approved and adopted the recommendations based on our recent collaborative study. The same modified method will be included in the Trading Rules of NCPA. (6) A quick and simple hydraulic press method was developed for the estimation of FFA in white cottonseed. It has been routinely used by a cottonseed mill in Georgia. 8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below) Wan, P.J., Wakelyn, P.J., Jividen, G.W. New Orleans researcher meets Chinese peers in Times Picayune. Downtown Picayune p. 4A2. February 15, 2001. Wakelyn, P.J., Wan, P.J. An overview of oils extraction and refining. Oil Mill Gazetteer. 2001. v. 106(8). p. 14-22.

Impacts
(N/A)

Publications

  • Wan, P.J. Properties of Fats and Oils. O'Brien, R., Farr, W., Wan, P., editors. Champaign, IL: AOCS Press; Chapter 2. 2000. p. 20-48. Introduction to Fats and Oils Technology.
  • Wan, P.J., O'Brien, R., Farr, W. Co-edited "Introduction to Fats and Oils Technology", 2nd edition. AOCS Press. 2000. p. 618.
  • Wakelyn, P.J., Wan, P.J. Food Industry - Solvents for extracting vegetable oils. Wypych, G. editor. Toronto, Canada and William Andrew Inc., Norwich, NY: ChemTec Publishing; Chapter 14. Handbook of Solvents. 2001. p. 923-949.
  • Wan, P.J., Britton, D. Effect of acid treatment of fuzzy cottonseed free fatty acid content determination. Proceedings of the 50th Oilseed Conference. 2001. p. W1-14.
  • Wan, P.J., Wakelyn, P.J., Jividen, G.W. China's cottonseed industries - advances and surprises. INFORM. 2001. v. 12(3). p. 236-243.
  • Wan, P.J., O'Brien, R.D. Cottonseed oil production and utilization. Lipid Technology. 2001. v. 13(3). p. 61-66.
  • Wan, P.J. Free Fatty Acids. Revised AOCS Official Method Aa 6-38. The 7th Edition of the Official Methods and Recommended Practices of the AOCS. Campaign, IL. 2001. p. 1.
  • Hron, R.J., Sr., Dowd, M.K. Oilseed and grain processing at the Southern Regional Research Center. Proceedings of the 50th Oilseed Conference. 2001. p. Q1-4.
  • Moreau, R.A., Singh, V., Hron, R.J., Sr., Hicks, K. A comparison of corn oils: corn germ oil, corn kernel oil, and corn fiber oil. Proceedings of the 50th Oilseed Conference. 2001. p. T1.
  • Wan, P.J., Pakarinen, D.R. Method to determine free fatty acid content in cottonseed: concerns and recommendations. INFORM. 2001. v. 11(5). Abstract p. S7.
  • Kuk, M.S., Hron R.J. Sr., Bland J.B. Gossypol complex with oilseed phospholipids. INFORM. 2001. v. 11(5). Abstract p. S110.
  • Dowd, M.K., Pelitire, S.M. Recovery of gossypol from cottonseed soapstock. Industrial Crops and Products. 2001. v. 14. p. 113-123.