1. Activated TAML Effects on Microbial Survivorship Ryan Kramer Grade 10 Pittsburgh Central Catholic

2. Problem Water systems are polluted with: Antibiotics Pharmaceuticals (Estrogen, etc.) Mercury Sulfur Sewage Fertilizers Organics Cleaning projects Dyes and other colored agents

3. Green Chemistry Chemistry designed to perform the same jobs as regular chemistry without the environmental hazards. Goals for Green Chemistry include: Eliminating or reducing atmospheric emissions. Finding an alternative to chlorine based water cleaners. Finding an alternative energy sources.

4. TAML GREEN catalyst engineered by Dr. Terry Collins at CMU Accelerates H 2 O 2 oxidation reaction Highly selective Made of common elements (C, H, O, N, Fe) GREEN Economical Easy to synthesize

5. TAML Continued Low working concentrations Degrades quickly Highly water-soluble Produces non-toxic byproducts Breaks down stable chlorine compounds among many others Practical uses include: Textile dye bleaching Pulp and paper bleaching Water cleaning Laundry stain remover Petroleum refining Rapid destruction of bio-chem warfare

6. Hydrogen Peroxide Pale blue liquid Used as disinfectant, antiseptic, and oxidizer Works with TAML in oxidation chemistry processes Weak acid Is the ideal green reagent: Nontoxic Inexpensive Disposable Fast acting

7. TAML and Hydrogen Peroxide Work together to clean water. TAML catalyzes (activates) peroxide reaction. Has been extensively tested and has effectively degraded numerous chemical pollutants. Is a safe, easy-to-use, inexpensive water disinfection technology.

8. Food Coloring Studies have shown colors affect taste For this reason companies add colors to enhance dining experience It is a non-toxic dye. Chosen because Its non-toxic Easy to obtain Serves as simple model of environmental color effluent problems

9. Saccharomyces Cerevisiae Used in baking and fermenting of alcoholic beverages. Most researched cell in the world. Serves as a simple model of microorganisms in aquatic ecosystems systems.

10. Catalase Common enzyme found in almost every living thing. Contains Porphyrin Heme, an iron group, that allows it to react with hydrogen peroxide. Functions as a catalyst for the decomposition of peroxide into water and hydrogen.

11. Purpose To determine if a quenched TAML/Peroxide reaction produces byproducts that are lethal to yeast cells.

12. Hypothesis Null Hypothesis: Quenched TAML/peroxide will have no significant effect on yeast survivorship. Alternative Hypothesis: Quenched TAML/peroxide will have a significant effect yeast survivorship.

13. Materials TAML Hydrogen Peroxide Red Food Coloring Sterile Pipettes 20mL Culture Tubes 58 YEPD agar plates (1% yeast extract, 2% peptone, 2% glucose (dextrose), 1.5% agar) YEPD Media (1% yeast extract, 2% peptone, 2% glucose (dextrose)) Saccharomyces Cerevisiae Sterile Filter Stock Solution Tubes Ethanol Bunsen Burner Spreader Bars Vortex Catalase Sterile Filter Distilled Water

14. Procedure A stock solution of red dye was generated (.015 uL dye per 10mL distilled water) in a stock solution tube. A 0.88M stock solution of hydrogen peroxide was generated in a stock solution tube. Food dye and water was sterilized with a 0.22 micron syringe and sterile filter.

15. Procedure Continued Water and Yeast Control Red Dye Control 2 Minute Quenched 20 Minute Quenched 40 Minute Quenched Yeast.1mL TAML0mL.5mL H202H202 0mL.5mL Red Dye0mL5mL Water9.9mL4.9mL3.8mL Catalase0mL.1mL

16. Liquid Pulse Exposure Tubes were vortexed to mix the ingredients. 100uL of the test subjects were spread on 8 agar plates per time period (4 per tube). All plates were incubated for two days. All colonies were counted.

17. Prolonged (agar) Exposure 0.5mL of the stock from the culture tube was mixed with 100uL of just water and yeast on 6 plates (3 per tube). All plates were incubated for two days. All colonies were counted. Water and Yeast Control Red DyeContinuous Reaction Yeast.1mL0mL TAML0mL.5mL H202H202 0mL.5mL Red Dye0mL5mL Water9.9mL5mL3.9mL Catalase0mL

18. Prolonged Agar Exposure Effect P Value=.11

19. Liquid Pulse Exposure W+D P Value= 4.6E-12 W+Y P Value=1.69E-10

20. Dunett’s Test for Water and Dye T Crit= 4.00 Extent of ExposureT ValueInterpretation 2 min.4.87Significant 20 min.7.75Significant 40 min.12.80Significant

21. Dunett’s Test for Water and Yeast T Crit= 4.00 Extent of ExposureT ValueInterpretation 2 min.10.73Significant 20 min.7.87Significant 40 min.2.86Insignificant

22. Conclusion All the exposure times varied significantly from the water and dye control group. Null Rejected At 2 minutes and 20 minutes the growth was significantly less than the water and yeast control group’s growth. Null Rejected The 40 minute group did not vary significantly from the water and yeast control. Null Accepted

23. Conclusion Continued Very little growth happened during the Prolonged Exposure Test. Null Accepted No significant variance from the control. Show that over a long period of time and increased concentration the byproducts of a quenched TAML/Peroxide reaction harms microbial life.

24. Extensions and Limitations Limitations: TAML is not specifically designed for food coloring. Yeast was just a model used for aquatic microbial life. Extensions: Use chlorine or other chemicals that are problems in aquatic ecosystems. Use a form of microbial life present in an aquatic ecosystem.

25. References http://www.chem.cmu.edu/groups/collins/ab out/about.html http://www.chem.cmu.edu/groups/collins/ab out/about.html http://en.wikipedia.org/wiki/Food_coloring http://en.wikipedia.org/wiki/Hydrogen_peroxi de http://en.wikipedia.org/wiki/Hydrogen_peroxi de http://en.wikipedia.org/wiki/Spectrophotome ter http://en.wikipedia.org/wiki/Spectrophotome ter http://en.wikipedia.org/wiki/Saccharomyces_ Cerevisiae