1. The Effects of Capsaicin on Microbes
Jacob Warden
Freshman
Central Catholic High School

2. Capsaicin
Capsaicin is a chemical irritant to all mammals that is found naturally in chili peppers.
Capsaicin is a naturally occurring chemical found in chili peppers, especially the flesh around the seeds.
Used as a medicine in diluted forms.

3. Chemistry of Capsaicin
Molecular FormulaC18H27NO3

4. Escherichia coli
Rod shaped bacteria found in the lower intestines of mammals
Some strains beneficial while others are pathogenic
Serves as a common prokaryotic cell model

5. Previous Studies
Journal of Ethnopharmacology 1999: Antimicrobial properties of alkamides present in flavoring plants traditionally used in Mesoamerica: affinin and capsaicin.

6. To determine what effect capsaicin extract has on E. coli survivorship
Purpose
To determine what effect capsaicin extract has on E. coli survivorship

7. Hypotheses
Null hypothesis: Capsaicin extract will not significantly affect the survivorship of E. coli.
Alternative hypothesis: Capsaicin extract will significantly affect the survivorship of E. coli.

8. Materials LB agar plates Escherichia coli
Capsaicin Extract (liquid form)
Micro and Macro pipettes + Sterile tips
Spreader bars
LB agar plates (1% Tryptone, 0.5% Yeast extract, 1% NaCl)
Escherichia coli DH5-alpha (E.Coli) (Obtained from Doonan Lab, CMU)
Bunsen Burner
Sidearm Flask
Vortex
Incubator
Gloves/goggles
Klett Spectrophotometer
SDF (Sterile Dilution Fluid)(100mM KH2PO, 100mM K2HPO4, 10mM MgSO4, 1m NaCl)
Sterile Test Tubes
Ethanol

9. Procedure Escherichia coli was grown overnight in sterile LB media.
A sample of the overnight culture was added to fresh media in a sterile sidearm flask.
The culture was incubated at 37°C until a density of 50 Klett spectrophotometer units was reached. (Cell density approximately 10^8 cells/mL).
The culture was diluted in a sterile dilution fluid to a concentration of approximately 10^5 cells/mL.
The selected experimental variables were diluted with sterile dilution fluid to the chosen concentrations to a total of 9.9 mL.

10. Procedure continued
mL. of cell culture was then added to the test tubes, yielding a final volume of 10 mL.
7. The solution was mixed by vortexing.
8. After vortexing to evenly suspend cells, 0.1 mL aliquots were removed from the tubes and spread on LB agar plates.
9. The plates were incubated at 37°C for 24 hours.
10. The resulting colonies were counted. Each colony is assumed to have risen from one cell.

11. Table of Concentrations
Column1
Tube 1
Tube 2
Tube 3
Tube 4
Tube5
Capsaicin Variable X
0ml
0.001ml
0.01ml
0.1ml
1ml
Sterile Fluid
9.9ml
9.899ml
9.89ml
9.8ml
8.9ml
Microbe
Final Volume
10ml
Final Concentration X 0%
0.01%
0.10% 1%
10%

12. The Effects of Capsaicin on Microbes
P-value:

13. T-Critical = 3.90 (significant)
Dunnett’s Test
T-Critical = 3.90 (significant)
Alpha = .05
Concentration of Capsaicin Extract
T-Value
Significant?
0.01%
 1.91 No
0.1%
 0.58 1%
 2.36
 No
10%
4.73 
 Yes

14. Conclusion
The null was rejected for the 0%, 0.01%, 0.1%, and 1% concentrations.
The null failed to be rejected for the 10% concentration.
The results and the statistical analyses indicate that 10% capsaicin extract did adversely affect E. coli survivorship.

15. Limitations The spread plating was not perfectly synchronized
One exposure time
Only one type of bacteria was tested
Limited number of concentrations tested

16. Extensions Different types of bacteria would be tested
More concentrations of the capsaicin extract would be tested
Synergistic effects would be tested
Multiple exposure times would be used

17. References
CSID: , Structure html (accessed 20:45, Jan 27, 2016) COPY
"Capsaicin." - Wikipedia, the Free Encyclopedia. N.p., n.d. Web. 22 Jan
"Escherichia Coli." - Wikipedia, the Free Encyclopedia. N.p., n.d. Web Jan
"Antimicrobial Properties of Alkamides Present in Flavoring Plants Traditionally Used in Mesoamerica: Affinin and Capsaicin." Antimicrobial Properties of Alkamides Present in Flavoring Plants Traditionally Used in Mesoamerica: Affinin and Capsaicin. N.p., n.d. Web. 27 Jan

18. Data Replicants #1 #2 #3 #4 Tube 1: 0% 248 300 244 212 Tube 2: 0.01%
352
392
180
408
Tube 3: 0.1%
166
280
174
284
Tube 4: 1%
172
112
132
Tube 5: 10% 17 65 28 81
Concentrations
Averages 0%
251
0.01%
333
0.10%
226 1%
149.5
10%
47.75

19. ANOVA One-Way ANOVA SUMMARY Group Count Sum Average Variance C1 4 1004
251 C2
1332
333 C3
904
226 C4
598
149.5 C5
191
47.75
ANOVA
Source SS df MS
Ftest
P-value
F-Critical
Between
186743
Within 15
Total
242197 19
Conclusions
Ho:
μ1=μ2=μ3=... Ha
At least 2 μ's differ
F-Test
Alpha
0.05
P-Value
Decision:
Reject Ho