Preventing Nicotine Toxicity

By: Kayleigh Makoid

Nicotine Epinephrine Daphnia Previous testing

Background

Are there inhibitory effects of epinephrine on nicotine toxicity?

Question

My hypothesis is if the nicotine concentration increases, then the daphnia

will die for both molar amounts of epinephrine.

Hypothesis

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

Materials

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= 10mM3. 1mL of above solution + 19mL of H20= .1mM4. 1mL of above solution + 9mL of H20= .01mM5. 1mL of above solution + 9mL of H20= .001mM6. 1mL of above solution + 9mL of H20= .0001mM7. Insert each solution into five separate 1.5 mL centrifuge

tubes

1. 4.5 mL of H20 in 3 flasks2. Add .5mL of epinephrine to first flask3. Add .5mL of the above solution to the second flask4. 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

Experimentation

heart

.1 Epinephrine solution12 Daphnia

2 in 1000mM

2 in 10mM

2 in .1mM

2 in .01mM

2 in .001mM

.001 Epinephrine solution12 Daphnia

2 in 1000 mM

2 in 10mM

2 in .1mM

2 in 1mM

2 in .001mM

Nicotine (mM) Heartbeats per min

1000mM 78, 8410mM 101, 98.1mM 107, 102.01mM 106, 112.001mM 104, 110

Tables .001M Epinephrine

Nicotine (mM) Heartbeats per min

1000mM 103, 10610mM 111, 109.1mM 114, 112.01mM 123, 131.001mM 160, 143

.1M Epinephrine

Heartbeats with H20

142, 151

.1M Epinephrine .001M Epinephrine

158, 154 157, 164

Nicotine (mM) Heartbeats per min

1000mM 81, 8710mM 94, 97.1mM 109, 98.01mM 110, 107.001mM 108, 104

Controls

Effect of Nicotine & Epinephrine on Heartbeat

IC50 for Nicotine=4.9990 Standard Deviation for Nicotine= 2.3604IC50 for Epinephrine=10.9100 Standard Deviation for Epinephrine=1.6994

Effect of Different Epinephrine Concentrations

IC50 for .001M epinephrine competition 27.9300IC50 for .1 M epinephrine competition 6.3520 x 10-

3

Standard deviation for .001 M is 0.2635Standard deviation for .1 M is 0.2187

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

Additional Information

Epinephrine is effective at .1M concentration to treat low dosages of Nicotine poisoning

Hypothesis partially supported

Conclusion

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

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“Epinephrine Injection.” MedlinePlus. N.p., 1 Sept. 2008. Web. 25 Jan. 2011. <http://www.nlm.nih.gov/ medlineplus/ druginfo/ meds/ a603002.html>.

Meeker-O’Connell, Ann. “How Nicotine Works.” Discovery Health. N.p., 2011. Web. 25 Jan. 2011. <http://health.howstuffworks.com/ wellness/ drugs-alcohol/ nicotine.htm>.

“Nicotine Information.” Home Drug Testing Kit. N.p., 10 Dec. 2002. Web. 25 Jan. 2011. <http://www.homedrugtestingkit.com/ nicotine.html>.

Scott, Elizabeth. “The Definition Of Epinephrine.” About. com. N.p., 22 Dec. 2007. Web. 25 Jan. 2011. <http://stress.about.com/ od/ stressmanagementglossary/ g/ Epinephrine.htm>.

Bibliography