Jacob Glaspey Nathan Horter Increasing the Yield of Fat Extracted from Processed Foods With Petroleum Ether Jacob Glaspey Nathan Horter

Fat Extraction Methods Laursen and Mernitz (1) – petroleum ether Folch Method (2) – chloroform-methanol Super Critical Fluid (3) There are several methods available for analyzing the amount of fat in a sample. The Laursen and Mernitz method is the method that we used in class earlier this quarter. This method uses petroleum ether as a solvent. The Folch Method was developed in 1957 by Jordi Folch for extracting lipids from brain tissue. This method has been used extensively for extracting lipids from animal carcasses and food products. Super Critical Fluid extraction is a relatively new method that uses supercritical carbon dioxide to extract the lipids from samples. We are focusing our studies on improving the extraction from the petroleum ether method. We also look to the Folch method for inspiration for our experimental methods. We will not be investigating supercritical fluid extraction as it involves a complicated hazardous method.

Method Advantages Lower Toxicity Lower Cost Less Complicated Petroleum Ether Chloroform-Methanol Lower Toxicity Lower Cost Less Complicated Quantitative Well Tested Less Time Required for Extraction So, lets look at the advantages of the petroleum ether extraction verses the chloroform methanol extraction. Petroleum ether has the advantages of lower toxicity, lower cost, and a less complicated extraction method. Chloroform-methanol is a quantitative method that is well tested. The extraction time is also a little less than that of the petroleum ether extraction.

Method Disadvantages Low Yield Time Intensive More Toxic Higher Cost Petroleum Ether Chloroform-Methanol Low Yield Time Intensive More Toxic Higher Cost Now lets look at the disadvantages. Petroleum ether, without an extraction apparatus, has a somewhat low yield, and requires more time to extract. Chloroform-Methanol is a toxic solvent that can cause cancer and has a higher cost than petroleum ether.

Goals of Our Investigation We want to: Increase the yield of petroleum ether extraction. Keep the complexity, cost, and hazards close to those of the control method. Be able to use our new method as an improved educational method. When we started out experimentation we had several goals in mind: We wanted to increase the yield do of the petroleum ether extraction. But we did not want to increase the complexity, cost, or hazards of the method. We put these restrictions on our experimental method because we wanted to be able to use our new method as an improved educational method. If the experiment became too complex, expensive, or hazardous beginning chemistry students could not perform it.

Control The volume of the petroleum ether solvent is 10 mL. The sample was extracted in the solvent for 1 hour. The extract was then dried overnight and weighed. So, before we can talk about our experimental alterations we need to go over the control method. As you may recall from earlier this quarter, the extraction uses 10 mL of petroleum ether to dissolve the fat in the sample. The sample was extracted for one hour with heat and agitation and then the excess solvent was boiled off and the extract was dried overnight. Once the extract was dry it was weighed and the percent fat was determined.

Our Experiments Experiment A – We increased the length of time of extraction. Experiment B – We increased the volume of solvent used. Experiment C – We altered the Folch Method by using petroleum ether instead of chloroform-methanol So, now we’re going to talk about our experimentations. As we started out we wanted to test if increasing the length of time of extraction or increasing the volume of solvent would increase the amount of fat extracted form the sample. In our last experiment we wanted to adapt the Folch method to our petroleum ether extraction to see if it would significantly increase the yield of the extraction.

Experiment A The length of time of extraction was increased from 1 hour to 2 hours. This experiment was designed to determine if the rate of the fat being pulled into solution was the limiting factor of the control. In experiment A we increased the extraction time form

Experiment B The volume of the solvent was increased from 10 mL to 20 mL. This experiment was designed to determine if the saturation of the solvent was the limiting factor of the control.

Experiment C The volume of the solvent was increased from 10 mL to 20 mL. The solvent and sample were then washed after the extraction with 4 mL of de-ionized water. This experiment was designed to determine if the procedure of the Folch method could improve the yield of our extraction.

Hey, is this going anywhere???

I’m getting bored.

Percent Yield of Total Fat Content by Method Our Results Percent Yield of Total Fat Content by Method Method Mean % Yield  95% confidence level Control 38  8 Experiment A 15  20 Experiment B 49  10 Experiment C 59  4

Statisticawhatsit? We calculated whether our experiments produced significant increases over our control using a two sample t test. The t value from these tests gave us a value of P. P is the probability that the two results are the same due to random variation.

Significance of Results Experiment B increased the yield by 11%. (P<.005) Experiment C increased the yield by 21%. (P<.001)

Future Experiments Drying the sample before extraction We’ll try: Drying the sample before extraction Using an acid digest on the sample before extraction Testing other low toxicity solvents Incorporating an extraction apparatus

Soxhlet Extractor Known to increase yield to > 90%. Classroom availability? A little expensive. VERY time consuming. (Around 16hrs for pet ether.) camel.campbell.edu/~chemistry/ faculty/jung/

Sonicawhatsit? It’s SONOCHEMISTRY. It (Jon, that is) may be of use to us. Sonication of the sample to decrease particle size may increase yield. Emulsifying the pet ether with the sample by sonication may increase yield. It may cause chemical reactions that break down fat. Nonetheless… Please Jon, help us.

OK, WE’RE DONE.

AT LAST!

Resources Laursen, Sandra; Mernitz, Heather. “Would You Like Fries With That? The Fuss About Fats In Our Diet”. John Wiley & Sons, Inc. 2000. New York. Folch, J.; Lees, M.; Sloane-Stanley, G.H. A simple method for the isolation and purification of total lipids from animal tissue. Journal of Biological Chemistry. 1957, 226, 497-509. Eller, Fred J.; King, Jerry W. Supercritical Fluid Extraction of Fat from Ground Beef: Effects of Water on Gravimetric and GC-FAME Fat Determinations, Journal of Agricultural and Food Chemistry; 2001; 49(10); 4609-4614.

Any QUESTIONS?