By: Patrick Leech
Pittsburgh Central Catholic
PJAS 2010
Grade 10
Entry into the water system
 Many pharmaceuticals enter the water through human
excretion.
 Direct disposal through household plumbing
 Water treatment plants only remove about 60% of the
drugs.
Benadryl
 Used as an antihistamine
 Works by blocking the effect of histamine receptor
sites
 Active Ingredient:
 Diphenhydramine hydrochloride
 Does not have FDA safe to flush label on it
 Trace amounts have been found in drinking water in
parts of the country
 Over 25 common over the counter drugs contain the same
active ingredient of Benadryl (Diphenhydramine
hydrochloride)
Ibuprofen
 A non-steroidal anti-inflammatory drug
 Used for relief of symptoms of arthritis, fever, and joint





pains
3rd most consumed drug in the world
About 30% of the amount of Ibuprofen bought enters
the water
Studies have shown that even low concentrations have
been found to affect the way fish spawn.
As Ibuprofen decomposes, it becomes more toxic.
Does not have FDA approved “safe to flush” label on it
Synergy?
 The two medicines Benadryl and Ibuprofen mixed
together
Algae
 Large diverse group of simple and usually
autotrophic organisms
 Base of aquatic food chain
 Used as a bio-indicator for
aquatic environments
Euglena
 Generally cylindrical in shape with many flagella
 Common algal like protist
 A partial heterotroph can attain food through
endocytosis and photosynthesis
 Nutrient-rich freshwater or in sewage systems
 Capable of survival in both salt and water
environments
 Frequently employed as an experimental
model
Purpose
 To discern the effect(s) of various concentrations of
Ibuprofen, Benadryl, and a combination of both on the
growth/survivorship of Euglena
Hypothesis
 Null: The presence of Benadryl, Ibuprofen, and a
combination of both will NOT have a significant effect
on algal growth.
 Alternative: The presence of Benadryl, Ibuprofen, and
a combination of both WILL have a significant on algal
growth.
Materials
 10% Benadryl solution (10 grams Benadryl per 100mL









Spring water)
10% Ibuprofen solution (10 grams Benadryl per 100mL
Spring water)
Euglena gracilis
Soil water (sterile)
Spring water
90 test tubes (13 x 100 mm borosilicate culture tubes)
Pipettes (macro + micro)
Pipette tips
Science Kit Educator spectrophotometer
Test tube rack
Procedure
A desk lamp was placed in experimental site
a)
approximately 45 centimeters away from the test tube racks
b) Kept on a 12 hour on 12 hour off cycle
c)
Kept at 22 degrees celsius throughout the duration of the study
2. Created a 10% stock solution of Ibuprofen, Benadryl, and a
combination of both
3. Created the concentrations of 0%, 0.01%, and 0.1% for Ibuprofen,
Benadryl, and the combination
1.
a)
4.
There were ten replicates per concentration
Readings were taken using a spectrophotometer set with a
wavelength of 430 nm once a day on days 1-10
Test Tube Ingredients
Concentration
0%
0.01%
0.1%
Algae
2mL
2mL
2mL
Distilled Water
3mL
2.995mL
2.95mL
10% stock solution
0mL
0.005mL
0.05mL
Total
5mL
5mL
5mL
Benadryl Influence on Euglena Population Growth
0.26
Absorbance at 430 nm
0.255
0.25
0.245
0.24
0% concentration
0.235
.01% concentration
0.23
1
2
3
4
5
6
Day
7
8
9
10
.1% concentration
Ibuprofen Influence on Euglena Population
0.26
Absorbance at 430 nm
0.255
0.25
0.245
0.24
0% concentration
.01% concentration
0.235
.1% concentration
0.23
1
2
3
4
5
6
Days
7
8
9
10
Synergy Influence on Euglena Population
0.26
Absorbance at 430 nm
0.255
0.25
0.245
0% concentration
0.24
.01% concentration
.1% concentration
0.235
1
2
3
4
5
6
Day
7
8
9
10
Results
 Is algal growth affected by the presence of Benadryl or
Ibuprofen in its environment?
 Ibuprofen?

P value-0.167411
NOT SIGNIFICANT
 Benadryl?

P value-0.098718 NOT SIGNIFICANT
 Synergy

P value- 0.08566 NOT SIGNIFCANT
Alpha = .05
Conclusions
 The null hypothesis was supported because any
variation that occurred was insignificant according to
the ANOVA statistical analysis
 After reviewing the statistical analysis the P value for
all three variables were greater than the standard cut
off of .05
 Because the P-value was greater than .05 on all
concentrations a Pair-wise test (Dunnett’s Test) was not
needed
Limitations and Extensions
 Limitations
 Euglena not fresh (Used 4 days after arrival)
 Extensions
 Use more diverse groups of algae
 Use a higher concentration of the drugs
 Use different kinds of drugs both over the counter and
prescription
 Use hemocytometer to obtain more accurate results
References
 www.medicinenet.com
 http://www.physorg.com/news120927003.html
 http://www.usa.siemens.com/entry/en/water.htm?stc
=usccc020156
 http://www.examiner.com/x-13868-Health-andScience-Examiner~y2010m1d19-Benadryl-Motrin-andRolaids-added-to-recall-of-Tylenolmakers-products