By: Kayleigh Makoid

 Nicotine  Epinephrine  Daphnia  Previous testing

 Are there inhibitory effects of epinephrine on nicotine toxicity?

 My hypothesis is if the nicotine concentration increases, then the daphnia will die for both molar amounts of epinephrine.

 Microscope  100–1000 μL micropipette  Pipettes  Distilled Water  Nicotine solution  Epinephrine  15 mL centrifuge tubes  1.5 mL centrifuge tubes  Stand for centrifuge tubes  Goggles  Gloves

Preparation of the Nicotine Solution Preparation of the Epinephrine 1. 2mL of Nicotine + 8mL of H20= 1M (1000mM) 2. 1mL of above solution + 19mL of H20= 10mM 3. 1mL of above solution + 19mL of H20=.1mM 4. 1mL of above solution + 9mL of H20=.01mM 5. 1mL of above solution + 9mL of H20=.001mM 6. 1mL of above solution + 9mL of H20=.0001mM 7. Insert each solution into five separate 1.5 mL centrifuge tubes mL of H20 in 3 flasks 2. Add.5mL of epinephrine to first flask 3. Add.5mL of the above solution to the second flask 4. Add.5mL of the above solution to the third flask

 Allow 12 daphnia to soak in each solution of epinephrine prepared for 10 minutes  Put daphnia under microscope  Add drop of each concentration of nicotine to different daphnia (two for every dilution of nicotine)  Count heartbeats for 1 min per Daphnia  Record results heart

.1 Epinephrine solution 12 Daphnia 2 in 1000mM2 in 10mM2 in.1mM2 in.01mM 2 in.001mM.001 Epinephrine solution 12 Daphnia 2 in 1000 mM2 in 10mM2 in.1mM2 in 1mM 2 in.001mM

Nicotine (mM) Heartbeats per min 1000mM78, 84 10mM101, 98.1mM107, mM106, mM104, M Epinephrine Nicotine (mM) Heartbeats per min 1000mM103, mM111, 109.1mM114, mM123, mM160, 143.1M Epinephrine Heartbeats with H20142, 151.1M Epinephrine.001M Epinephrine 158, , 164 Nicotine (mM) Heartbeats per min 1000mM81, 87 10mM94, 97.1mM109, 98.01mM110, mM108, 104 Controls

IC50 for Nicotine= Standard Deviation for Nicotine= IC50 for Epinephrine= Standard Deviation for Epinephrine= IC50 for Nicotine= Standard Deviation for Nicotine= IC50 for Epinephrine= Standard Deviation for Epinephrine=1.6994

IC50 for.001M epinephrine competition IC50 for.1 M epinephrine competition x Standard deviation for.001 M is Standard deviation for.1 M is IC50 for.001M epinephrine competition IC50 for.1 M epinephrine competition x Standard deviation for.001 M is Standard deviation for.1 M is

Independent variable  Concentration of epinephrine Dependent variable  Heartbeats of the Daphnia Controls  Water, epinephrine, and nicotine Constants  Room temperature, fume hood, microscope, and daphnia species

 Epinephrine is effective at.1M concentration to treat low dosages of Nicotine poisoning  Hypothesis partially supported

Factors the Affected Accuracy Future Experimentation  Death of Daphnia  Aging solutions .1M Epinephrine’s potency on Daphnia  Inability to be precise with counting  Test other molar amounts of epinephrine on nicotine

 Clare, John, P.. D, “Daphnia.” Caudata. N.p., July Web. 25 Jan  “Epinephrine Injection.” MedlinePlus. N.p., 1 Sept Web. 25 Jan  Meeker-O’Connell, Ann. “How Nicotine Works.” Discovery Health. N.p., Web. 25 Jan  “Nicotine Information.” Home Drug Testing Kit. N.p., 10 Dec Web. 25 Jan  Scott, Elizabeth. “The Definition Of Epinephrine.” About. com. N.p., 22 Dec Web. 25 Jan