1. TAML Remediation of Eutrophic Water
Simon Sweeney
Central Catholic High School
9th Grade

2. Problem
Water pollution harms the ecosystem, a solution must be found to combat the pollution by degrading the pollutants in water to negate harmful effects on organisms.
Pollution is a huge danger to the Earth's ecosystem.
Harmful chemicals common in polluted water include
Gasoline
Pesticides
Chlorine
Fertilizers

3. TAMLs
GREEN catalyst engineered by Dr. Terry Collins at Carnegie Mellon University
Accelerates H2O2 oxidation reaction
Highly selective
Made of common elements
(C, H, O, N, Fe)
Economical
Easy to synthesize

4. TAMLs (cont.) Low working concentrations (nM-low μM)
Breaks itself down after reaction
Highly water soluble
Produces non-toxic byproducts
Breaks down stable chlorine compounds
Step towards a sustainable society
Reduce exotic elements
Think in terms of nature’s chemistry

5. Possible Applications of TAMLs
●Textiles
●Pulp and Paper
●Water Cleaning
●Petroleum Refining
●Biological Warfare

6. Isopropyl Alcohol
Alcohol- the hydroxyl functional group is bound to a saturated carbon atom
Isopropyl Alcohol
A common rubbing alcohol, toxic
Formula: C3H8O

7. Escherichia coli
E. coli – very common, found in intestinal tract of most mammals
There are many strains of E. coli, most are non-pathogenic
Pathogenic strains can cause illness and death in humans
Frequently studied in biology

8. Purpose
Assess the success of TAML at degrading Isopropyl Alcohol by means of a biological assay

9. Hypotheses
Null: TAML will have no significant effect on the lethality of the Isopropyl Alcohol.
Alt: TAML will degrade Isopropyl Alcohol and negate the decrease in E. coli survivorship

10. Materials Isopropyl Alcohol (70%) Fe-TAML B catalyst H2O2 (.88 M)
Lab-Grade Catalase (Ward’s)
E. coli DH5α
Sterile 250mL sidearm flask
NaHCO3 (Store bought baking soda)
Sterile Dilution Fluid (10mM KH2PO4, 10mM K2HPO4, 1 mM NaCl)
Sterile test tubes
Micropipette and sterile tips
20-200uL pipette and sterile tips
Sterile Luria broth agar plates (1% Tryptone, 0.5% Yeast Extract, 1% NaCl)
Tube racks
permanent marker
Ethanol
Vortex
Incubator
Bunsen burner

11. Procedure
A culture of E. coli DH5α was grown overnight in a sterile 250mL sidearm flask in LB media at 37°C.
The culture was left to grow until it reached an absorbance reading of about 50 Kletts, which represents a cell density of about 108 cells/mL.
The culture was serially diluted in SDF to a concentration of approximately 103 cells/mL.
Petri dishes, poured with LB agar were labeled and left to warm on the lab desk.
0.0353g of solid Fe-TAML B was added to 10mL of sterile water to create a 5*10-3M stock solution.
0.89g of solid NaHCO3 was added to 10mL of sterile water to create a 1M stock solution.
0.1mL of solid lab-grade catalase was added to 30mL of sterile water to create a working concentration of catalase.

12. Procedure (cont.)
8. Concentrations of Isopropyl alcohol and H2O2 were added to 6 sterile test tubes containing SDF.
9. TAML activators were added to 3 of the tubes
10. Tubes were let sit for 20 minutes while the TAML oxidation reaction occurred
11. Catalase was added to all 6 of the tubes
12. Tubes were let sit for 15 minutes while catalase quenched the reaction
13. E. coli were added to all 6 of the tubes
14. Tubes were let sit for 20 minutes while the E. coli reacted to the solution
mL of each concentration was plated onto LB agar plates
16. Plates were incubated at 37°C overnight, and the resulting colonies counted

13. Concentrations (in mL)
0% Alcohol, No TAML
1% Alcohol, No TAML
0% Alcohol, with TAML
1% Alcohol, with TAML
Isopropyl Alcohol
0.1
SDF
9.1 9
8.9
E. coli
TAML
H2O2
0.5
Catalase
0.2
Sodium Bicarbonate
Total 10

14. Average Colony Survivorship without Quenched Reaction Mix
P-value=

15. Average Colony Survivorship with Quenched Reaction Mix
P-value=

16. Stats
Isopropyl Alcohol Effect on Survivorship Significant Quenched Reaction Mix and Alcohol Effect on Survivorship Not Significant

17. Interpretations
The solution of TAML Activated H2O2 with Isopropyl Alcohol did not have a significant effect on the survivorship of E. coli.
The Isopropyl Alcohol solution that did not include TAMLs did have a significant effect on the survivorship of E. coli.
The TAMLs appeared to successfully degrade the Isopropyl Alcohol.

18. Conclusions REJECTED ACCEPTED
Null: The Isopropyl Alcohol will have no significant effect on E. coli survivorship.
REJECTED
Null: The TAMLs will have no significant effect on the lethality of the Isopropyl Alcohol.
Alt: The TAML activated H2O2 will have a significant effect on the degradation of the alcohol.
ACCEPTED

19. Limitations Spread plating was inconsistent in timing and technique
Low range of Alcohol Concentrations, TAML and Peroxide
Only one exposure time
Only one type of exposure (liquid pulse)
Isopropyl alcohol not a common pollutant
Only one microbial species
Limited to one model
Limited to one toxic substance

20. Extensions Greater range in exposures and time of exposure
More microbial models
Different type of exposure
Chemically define the degradation products from the TAML reaction

21. References
Carnegie Mellon University Green Science
contamination/water-contaminants-pollutants-list.htm
ysts-remove-estrogenic-compounds-from-wastewater.html

22. Results (colony counts)
Replicant
0% Alcohol, No TAML
1% Alcohol, No TAML
0% Alcohol, with TAML
1% Alcohol, with TAML 1
141
109 86 60 2
152
101 87 71 3
175 96 83 72 4
126
111 79 5
121 56 70

23. ANOVA (No Quenched Reaction Mix)

24. ANOVA (Quenched Reaction Mix)