Temperature Change Influences the Viscosity Flow Rate of Saturated Fatty Acids in Dietary Supplementation
The purpose of this analysis project is to identify essential fatty acids found in dietary supplementations in Utah. Marketed products that contain less than the equal advertised claim disables consumers, patients, clinical and medical researchers.
The first phase of this project obtained different products, varying in cost and brand availability. This research also created a replicable method of producing these types of results. The final phase tested the viscosity of these samples with the change of temperature and examined the data while comparing known melting temperatures of DHA, EPA, and ALA.
By identifying the percent of viscosity change, relative to each sample, 60% of the products tested failed to meet the label claim. The ratios used by manufacturers also associated with cost and ratios of DHA and EPA. This research will allow for more significant consideration for consumers, researchers and doctors on the change of the dietary supplementation market.
NOTICE: The “general information” section has been moved to better fit the styles of this scientific report. More can be found on his link.
Methods or Methods and Materials
- The test in this study consisted of seven samples. All samples contained a label claim and purchased through local shopping stores. All samples remain kept at room temperature with no sunlight and unpackaged until the day of experimentation.
- Sample #1 is produced using essential oils and filtration, with the claimed label of 600mg of omegas, 324mg EPA, 216mg DHA. The ingredients include Alaska Walleye Pollock, Pacific Whiting essential oils and Mixed Tocopherols. Costs per soft-gel are $0.07.
- Sample #2 is produced by molecular distillation, with the known label of 130mg of omegas, 430mg of EPA, 390mg of DHA and in triglyceride form. The ingredients include anchovy, tocopherols, rosemary extract and ascorbic palmitate. Average cost per soft-gel is $0.85.
- Sample #3 Production method is not labeled, with the claimed label of 720mg of omegas, 360mg of EPA, 240mg of DHA. The ingredients include sardine, mackerel, sodium alginate, ammonium hydroxide, medium chain triglycerides, tocopherols, stearic acid, sunflower oil. Costs per soft-gel are $0.13.
- Sample #4 Production method is not labeled, with the claimed label of 600mg of omegas, 400mg of EPA and 200mg of DHA. The ingredients include anchovy, sardine, and tocopherols. Costs per soft-gel are $0.10
- Sample #5 Production method is not labeled, with the claimed label of 90mg of omegas, 50mg of EPA and 24mg of DHA. The ingredients include Krill, gelatin, glycerin, water, sorbitol. Costs per soft-gel are $0.66.
- Sample #6 Production method is not labeled, with the claimed label of 300mg of omegas, 700mg of unknown oil concentrate. The ingredients include anchovy, gelatin, glycerin, tocopherols. Costs per soft-gel are $0.05.
- Sample #7 is produced by purification, with the known label of 50mg of GLA and 369mg of LA. The ingredients include evening primrose oil (Seed). The cost per soft-gel is $0.55.
The determination of the compensation in each sample is calculated using the lowest temperature reached by the sample. Each sample in each test interval is tested three times. The number of milliseconds of drips is averaged by the minimum (lowest) drip rate number for each sample and multiplied by 70%. If the sample drip rate is not measurable, then if the sample is soft, soft is equivalent to 80% viscosity; then else if wax, the wax is the equivalence to 90% viscosity, then if froze, froze is the equivalence to 100% viscosity, else sample is noted and skipped from data. Test set interval is -1C l unless the temperature is -15C. If the temperature is -15C, then Test set interval is -5C.
The temperature is lowered by -1C intervals until reaching 5C. The sample is cooled for at least 5 minutes. Temperature is checked with the second thermostat. If the temperature reaches -15C, then the intervals are changed to -5C intervals.
Viscosity is measured by minimum drip rate number in the experiment unless the sample reaches a soft, waxy or frozen consistency. The viscosity was determined by analyzation of video recordings of 30 fps using video editing software. An iPad Pro 9.7” is used to record using default settings. Final cut pro is used to edit the video and is overlaid with a time counter video. This video is freeze framed after each sample is analyzed. The video is cut after the 3 second mark. No additional manipulation of the video is used. Titles are placed on samples that are too viscous to stir. The freeze frame setting is set to 3 seconds in settings menu. All data is placed on charts using excel. A line chart for each sample.
- 22qt. Insulated Styrofoam cooler (for dry ice)
- Video camera (30 fps)
- Styrofoam board
- 4 feet by 1 foot
- Snap-off utility knife
- Two glass panes (11×8 inches)
- Aluminum tape
- General masking tape
- Electrical tape
- 12v digital thermostat temperature control switch sensor module with the minimum temperature ranges of -50 to 110c
- 10 feet of electrical wiring (use caution, follow switch guidelines)
- Eight pounds of dry ice (use caution)
- Clear vials with lids
- 80mm cooling fan (insignias worked fine, until reaching -30℃)
- Wire strippers
- Probe thermometer
- Digital thermometer
- Alcohol 90% isopropyl alcohol
- 110v to 12v dc 5a 60w regulated switching power supply
A 22qt. Styrofoam cooler box was used for this experiment. A square hole was cut on the long side of the box, the size of the hole is 10×7 inches or less (See figure 1). One glass pane was placed in outside of the container and taped in place using scotch tape. The second pane was placed in the inside of the box, covering the hole. This created an air pocket that acted as an insulator. Scotch tape was used to seal and hold the windows into place.
A bottom Styrofoam plate was placed 2 inches above the bottom, supported by thin strips of Styrofoam. This play fit snuggled next to the two long walls of the box. After experimentation, three layers of scotch tape were added to the plate with one final layer of aluminum. The bottom plate has gaps on the sides to allow air circulation. A wall was taped to the inside of the box, 2 inches away from the side. The wall contains a fan that allows the movement of circulation. The digital was set to turn on and off at when the temperature increased by – / + 0.2 degrees C.
A second inside lid was created on top of the cooler with holes for tube access. The original lid can cover and seal if needed (see figures 4 and 5).Different types of Styrofoam boxes were used for this experiment. I found that the 2” thick wall sided Styrofoam hold temperate better than other types of Styrofoam. Aluminum tape transfers heat quickly and is not an ideal product for sealing purposes, for this project.
- A large dry ice piece is broken by a hammer and placed, under the Styrofoam plate. The inside lid is on, and tubes are placed through the holes created. The new scotch tape is used to cover the inside lid. Thermometers are inside a control specimen of omega 3, and both the thermostat and digital thermometer are placed in a solution of alcohol 90% isopropyl alcohol inside the box. The power was turned on.
- After a couple of minutes, the thermostat began switching on and off, repeating fast, indicating that temperatures are stabilized for the alcohol solution. Using the probe thermometers, the samples were checked. Then all samples were recorded using a video camera and tested. A plastic straw is used to carry the sample from the vial into the straw.
- The straw is plugged by one finger and is placed a distance away from the bottom of the container. The straw is unplugged, and the sample drops back into the container. This process is repeated and recorded until the samples no longer can drip from the straw. The viscosity as described in the scoring section were used to describe viscosity.
Each sample is repeated three times to obtain an average. If the sample is unable to drip, then the sample is stirred to check for one of three consistencies. Soft consistency is resembled cold honey and is unable to drip. Wax consistency resembles shortening or butter; it is soft enough to mold. Froze is the consistency of ice; a toothpick will be unable to penetrate the surface.
The time range for each sample begins one millisecond before the drop and one millisecond after the first drop. A plastic straw is used to carry the sample from the vial into the straw. The straw is plugged by one finger and is placed a distance away from the bottom of the container. The straw is unplugged, and the sample drops back into the container.
In a study containing 7 different omega marketed samples, containing fish oil, krill oil, flaxseed oil and evening primrose oil are tested for EPA, DHA, ALA or GLA. In this test, 57% of these samples failed to meet the label claim. 5 of 7 samples, 71% contained EPA and 3 of those samples, 43% contained DHA. 4 of the 7 samples, 57 % of the samples contained ALA. 4 of the 6, 66% of the samples contained GLA. 2 of the 5 samples, 40% of the samples failed to meet or contain DHA. 3 of the 5, 60% samples failed to meet or contain EPA. 1 additional sample failed to contain DHA, this sample claimed of containing varied Omegas. Sample #1 failed to contain either Linoleic Acid and Linolenic Acid. This determines that 40% of the products tested.
In further analysis, using negative and positive chart line flow rates indicate that all of the samples may contain Palmitoleic acid 100% of all samples, Nonmelodic acid at 50% of the samples, 40% Docosahexaenoic acid of all samples and 20% of Docosahexaenoic acid of all samples.
In further prospective analysis of costs and ratios of DHA and EPA, the cost average correlate with the ratio of claimed EPA and DHA. 2 samples that are less than $0.10 contain EPA and typically consists of a ½ or 2/3 ratio of EPA and DHA respectively. 2 samples cost greater than $0.10 per capsule and claim greater levels of EPA and DHA of 13/43 and 12/25 respectively. The samples show that greater ratio of DHA are greater cost and contain less than 75% of DHA. While 2/3 ratios of costs lower than $0.10 per capsule contain more EPA and DHA overall.
As a review. 60% of the samples contained a form of Linolenic acid, in either gamma or alpha form. 75% of the products contained Linolenic acid in GLA form. 20% of the products contained Linolenic acid in GLA form. 50 % of those samples contained Linoleic or Nonmelodic acid. 100% of these samples contained Palmitoleic acid. 40% of these samples contained Docosahexaenoic acid. 20% of these samples contained Arachidonic acid.
Results of All Samples
|Sample||Linoleic Acid (MP of -5℃)||Linolenic (MP of -11℃)||Arachidonic (MP of -50℃)||Other 1||Other 2||Other 3|
|#1||–||+||–||-1, -5||-17, -30|
|#2||+||+||–||-7, -9||-18, -30|
|#3||+||–||–||1, -1||-7, -8||-18, -30|
|#4||–||–||+||1, -1||-50 < X|
|#5||+||+||–||0, -2||-50 < X|
|#6||+||+||–||0, -1||-50 < X|
|#7||+||+||–||1, -1||-50 < X|
Changes based on other oils
|Lipid Numbers||TEMP °C||Common Name||#1||#2||#3||#4||#5||#6||#7|
In the dietary supplementations studied, A total of 60% of the products samples failed to meet the dietary claim of contain both EPA and DHA. 3 of the 5 products, 60%, failed to meet the dietary label claims of containing EPA, while 2 out of the 5, 40% failed to meet the label claim of docosahexaenoic acid. 1 product only EPA as marketed for containing “various Omegas”. 1 additional product failed to contain DHA, while meeting the label claim of containing GLA.
Costs and advertising are also briefly analyzed, determining that most of these products may contribute to marketing schemes. In two of the highest costing samples, the ratio of EPA to DHA mismatched the 2:3 ratios, while the costs of 2:3 are $0.7-$0.10 per capsule, these samples contained EPA and DHA.
This finding is concerning and calls for additional follow-up research and analysis review to determine the effectiveness of product label claims found in most common supper markets. It is also essential to educate the public and medical communities of the product label claims.
The application of these findings affects many ongoing claims, research studies, the FDA and the general public. Many studies report of little of effectiveness of omega-3 supplementation when physicians, cross links of those patients taking low or no dosage of the studied supplementation may play an essential outcome of the true nature and effectiveness other studies have found. This may also reduce the constraints the FDA has placed in the market for dietary oils, known as common food source dietary supplements. This overall will have a larger impact on patients.
- Albert, B. B., Derraik, J. G., Cameron-Smith, D., Hofman, P. L., Tumanov, S., Villas-Boas, S. G., . . . Cutfield, W. S. (2015). Fish oil supplements in New Zealand are highly oxidised and do not meet label content of n-3 PUFA. Sci Rep, 5, 7928. doi:10.1038/srep07928
- Anonymous (Producer). (2018). Anonymous.
- . AP 42, Fifth Edition, Volume I Chapter 9: Food and Agricultural Industries. In (Vol. AP 42, Fifth Edition, Volume I pp. 1-7). 1995: U.S. Environmental Protection Agency. Retrieved from https://www3.epa.gov/ttn/chief/ap42/ch09/final/c9s13-1.pdf.
- Eilat-Adar, S., Mete, M., Nobmann, E. D., Xu, J., Fabsitz, R. R., Ebbesson, S. O., & Howard, B. V. (2009). Dietary patterns are linked to cardiovascular risk factors but not to inflammatory markers in Alaska Eskimos. J Nutr, 139(12), 2322-2328. doi:10.3945/jn.109.110387
- Kleiner, A. C., Cladis, D. P., & Santerre, C. R. (2015). A comparison of actual versus stated label amounts of EPA and DHA in commercial omega-3 dietary supplements in the United States. J Sci Food Agric, 95(6), 1260-1267. doi:10.1002/jsfa.6816
- Nichols, P. D., Dogan, L., & Sinclair, A. (2016). Australian and New Zealand Fish Oil Products in 2016 Meet Label Omega-3 Claims and Are Not Oxidized. Nutrients, 8(11), 703. doi:10.3390/nu8110703
- Opperman, M., Marais de, W., & Spinnler Benade, A. J. (2011). Analysis of omega-3 fatty acid content of South African fish oil supplements. Cardiovasc J Afr, 22(6), 324-329. doi:10.5830/CVJA-2010-080
- Pigott, G. M., & Fisheries, U. S. B. o. C. (1967).Production of fish oil: U.S. Dept. of the Interior, Fish and Wildlife Service, Bureau of Commercial Fisheries.
- Singh, S., Arora, R. R., Singh, M., & Khosla, S. (2016). Eicosapentaenoic Acid Versus Docosahexaenoic Acid as Options for Vascular Risk Prevention: A Fish Story. Am J Ther, 23(3), e905-910. doi:10.1097/MJT.0000000000000165
- Spector, A. A., & Kim, H. Y. (2015). Discovery of essential fatty acids. J Lipid Res, 56(1), 11-21. doi:10.1194/jlr.R055095
- Supplements, O. o. D. (2018). Omega-3 Fatty Acids Fact Sheet for Health Professionals. Retrieved from https://ods.od.nih.gov/factsheets/Omega3FattyAcids-HealthProfessional/