Jill Fisher Science Seminar Dr. Heyen, Advisor Determining the Chemical Make-up of the Flavoring of Coca-Cola and the Degradation of Plastic Bottles Jill Fisher Science Seminar Dr. Heyen, Advisor

Purpose of Experiment, Part 1 To compare chemicals of proposed Coca-Cola recipe with chemicals found from Coca-Cola product using gas chromatography/mass spectroscopy and ultraviolet spectrophotometry.

Methods Extract organic compounds with ethyl ether, compare using gas chromatography/mass spectroscopy (GCMS), run spectrum using ultraviolet (UV) spectrophotometer Dilute with methanol, compare using GCMS Dilute with water, run spectrum with UV spectrophotometer

GCMS Coca-Cola samples dissolved in methanol or extracted with ethyl ether were heated and detected for comparison with chemicals in a database.

UV Spectrophotometry Extracted and diluted Coca-Cola samples were ran in a spectrum of 190 nm – 320 nm Diluted sample was tested with certain wavelengths for absorbency/%transmittance

Target Chemicals Ingredient Chemical Structure Wavelength (nm) Citric Acid --- Vanilla Vanillin 279, 309 Orange Oil Limonene 220, 250 Cinnamon Oil Eugenol Cinnamaldehyde 291 Caryophyllene 280 Linalool 243 Estragole 230, 280 Lemon Oil Coriander Oil See above Pinene 210 Nutmeg Oil Myristicin Elemicin Sabinene Terpinene 265 Safrole 236, 285 Neroli Oil Nerol 189-190

Results from GCMS The GCMS analysis shows the presence of several peaks, thus several chemicals No exact matches, but possible derivatives of target chemicals Example: Vanillin Vanilglycolic acid

Myristicin/Safrole derivative Vanillin derivative Myristicin/Safrole derivative

Results from Spectrophotometer Wavelength (nm) ABS/%T Target 309 -0.03/136 vanillin 291 0.40/41.3 ☺cinnamaldehyde 285 0.82/15.2 safrole 280 1.40/4.0 estragole/caryophyllene 279 1.48/3.2 265 2.33/0.5 terpinene 250 3.01/0.1 limonene 243 3.10/0.1 ☺linalool 236 2.60/0.3 230 2.30/0.5 estragole 220 1.80/1.5 210 0.92/15.0 pinene 194 0.40/34.0 nerol 192 0.33/42.0 190 0.10/100 Spectrum gave peaks for cinnamaldehyde and linalool Several targeted wavelengths formed one peak Photometer gave individual absorbencies for entered wavelengths

Target Chemicals Ingredient Chemical Structure Wavelength (nm) Citric Acid --- Vanilla Vanillin 279, 309 Orange Oil Limonene 220, 250 Cinnamon Oil Eugenol Cinnamaldehyde 291 Caryophyllene 280 Linalool 243 Estragole 230, 280 Lemon Oil Coriander Oil See above Pinene 210 Nutmeg Oil Myristicin Elemicin Sabinene Terpinene 265 Safrole 236, 285 Neroli Oil Nerol 189-190

Conclusions The secret flavoring of Coca-Cola does include citric acid, vanilla, and cinnamon oil. Coriander and nutmeg oil have a high possibility of being included. Recipe may have changed over the decades.

Purpose of Experiment, Part 2 To test for the presence of acetaldehyde in water contained in plastic bottles under different environments for an extended period of time. heat Polyethylene terephthalate (PETE) Acetaldehyde

2,4-dinitrophenylhydrazine Methods Extract organic compounds with ethyl ether React aldehyde with 2,4-dinitrophenylhydrazine Extract organic compounds with Solid Phase Extraction tubes Identify with GCMS 2,4-dinitrophenylhydrazine

2,4-dinitrophenylhydrazine (DNPH) 2,4-DNPH reagent is made with phosphoric acid, ethyl ether, and 2,4-DNPH The DNPH will react with aldehydes to give 2,4-dinitrophenylhydrazone

Solid Phase Extraction (SPE) Water is pulled through column with suction filtration. Chemicals on media are eluted with methanol

Sample Environments Water stored for 36 days in Greenhouse, Organic Lab, and Microbiology Refrigerator Temperatures of water at collection: Greenhouse: 29.5°C Organic Lab: 20.5°C Refrigerator: 6.0°C

Solid Phase Extraction Target Chemicals DNPH Reaction Solid Phase Extraction 2,4-dinitrolphenylhydrazone 2,4-dinitrolphenylhydrazine Ethyl ether Acetaldehyde Methanol

Results from GC Samples mixed with DNPH reagent did not give any noticeable results Samples from SPE only showed excess chemicals from greenhouse container

Greenhouse sample with DNPH Organic lab sample with DNPH Fridge sample with DNPH

Greenhouse sample from SPE Organic lab sample from SPE Fridge sample from SPE

Conclusion The DNPH did not show the presence of acetaldehyde. The SPE did show a presence of a larger compound, perhaps from the PETE or from the silica gel matrix.

Acknowledgements and Thanks Thanks to Dr. Heyen for being my advisor and helping with the equipment and procedure. Thanks to Diane Soldan and Caleb Mason for chugging Coca-Cola.

Sources 2,4-dinitrophenylhydrazine. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/2%2C4-dinitrophenylhydrazine Acetaldehyde. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Acetaldehyde Cinnamon. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Cinnamon Coca-Cola. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Coca-Cola_formula Coriander. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Coriander Neroli. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Neroli Nutmeg. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Nutmeg Orange Oil. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Orange_oil Polyethylene Terephthalate. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Polyethylene_terephthalate The Coca-Cola Recipe. Soda Museum. http://www.sodamuseum.bigstep.com/generic.jhtml?pid=10 The Safety of Convenience-Size Plastic Beverage Bottles. Plastics Info. http://www.plasticsinfo.org/beveragebottles/apc_faqs.html Van Aardt, Marleen. “Effect of Shelf-Life and Light Exposure on Acetaldehyde Concentration in Milk Packaged in HDPE and PETE Bottles.” Virginia Polytechnic Institute and State University, 2000. Vanilla. Wikipedia Free Encyclopedia. http://en.wikipedia.org/wiki/Vanilla Weast, Robert C, ed. CRC Handbook of Chemistry and Physics. 57th Ed. Cleveland: CRC Press, 1976. Williamson, Kenneth. Macroscale and Microscale Organic Experiments. 3rd Ed. Boston: Houghton Mifflin, 1999. Zubrick, James W. The Organic Chem Lab Survival Manual. New Jersey: John Wiley & Sons, Inc. 2004.