TAML Process Effluent Effect on Aquatic Productivity

Joe DiPietro Central Catholic High School

11th grade

Developed and engineered by a research group led by Dr. Terry Collins of Carnegie Mellon University

Accelerates Hydrogen Peroxide(H2O2) reaction

Made of common elements found in nature (C, H, O, N, Fe)

◦ Environmentally biodegradable and

safe

◦ Economical

◦ Easy to make

Requires low concentrations

to activate

Tetra Amido Macrocyclic Ligand(TAML)

Central iron atom, where hydrogen peroxide bonds

Attached to a ringed carbon structure (Macrocylic) by 4 nitrogen atoms(TetraAmido)

Structure of TAML

Water purification

Catalyze the breakdown of pesticides

Pulp and paper ◦ Purify the effluent from paper mills

Petroleum refining ◦ Eliminates corrosive dibenzothiopes

Applications of TAML

Local attraction during all times of the year by joggers and families

Large duck population and other organisms

Lots of pollution and trash

Unhealthy environment

Primary location for algal growth

Panther hollow pond

Algae is an essential part of the ecosystem as it produces around 20% of atmospheric oxygen

Produces the majority of the oxygen utilized by aquatic ecosystems

Main source of food for many fish and other aquatic organisms

Importance of Algae

Determine if the effluent of the TAML process has an effect on algal growth and productivity in panther hollow pond

Determine if there is pollution in Panther

hollow pond

Purpose

Null-TAML will not have a significant effect on the productivity of algae in panther hollow pond, quenched or unquenched

Alternative-TAML will significantly improve algal productivity by reducing pollution levels in panther hollow pond without harming algae

Hypotheses

La motte dissolved oxygen titration kit ◦ 6 dissolved oxygen glass sample bottle

◦ 30 mL Manganous sulfate solution

◦ 30 mL Alkaline Potassium iodide

◦ 30 mL Sulfuric acid

◦ 30 mL Starch indicator solution

◦ 60 mL Sodium Thiosulfate

◦ Direct reading titrator

Pipettes

Sterile tips

Pond water from panther hollow pond

Light source (13 watt fluorescent bulb)

TAML 5 x 10^-3 stock solution

Hydrogen Peroxide

10 mg/mL of catalase

Materials

Productivity is the amount of biomass produced in an ecosystem

Measured by the dissolved oxygen produced ◦ Directly indicates if more biomass has been created

◦ Commonly accepted method for measuring dissolved oxygen (Winkler method)

◦ mL of Oxygen produced x 0.536=mg of carbon fixed

Measuring productivity

Added Manganous sulfate and Alkaline Potassium Iodide Azide solution to form a precipitate of Manganous hydroxide

◦ MnSO4 + 2KOH --> MN(OH)2 + K2SO4

Oxygen in the water oxidized the Manganous Hydroxide to form Manganic Hydroxide which was a brown precipitate

◦ 4MN(OH)2 + O2 + H2O --> 4MN(OH)3

Testing dissolved oxygen

A strong acid was added to the sample to “fix” it.

◦ 2MN(OH)3 + 3H2SO4 --> Mn2(SO4)3 + 6H2O

Iodine from the Potassium Iodide solution was oxidized by the Manganous Sulfate which released free iodine

◦ Mn2(SO4)3 + 2KI --> 2MnSO4 + K2SO4 + I2

Added Sodium Thiosulfate which reacts with the iodine to produce Sodium Iodide which changed the color of the solution from yellow to clear ◦ 2Na2S2O3 + I2 --> NaS4O6 + 2NaI

Testing dissolved oxygen cont.

Gathered 6 water samples in bottles without excess oxygen ◦ Tightly caped bottle underwater ◦ Unscrewed underwater ◦ Tapped bottle to release bubbles on the side of the bottle

and re-cap

Added activated TAML to second sample and allowed reaction to occur for 2 minutes

After 2 minutes added 0.01 mL of catalase to quench the reaction

Added TAML to third sample and allowed reaction to run its course

Procedure

Exposed samples to light source for 12 hours at a distance of 18 inches

Performed dissolved oxygen test on control group

Performed dissolved oxygen test on quenched TAML group

Performed dissolved oxygen test on unquenched TAML group

Analyzed results

Procedure cont.

Effects of TAML effluent on aquatic productivity

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

16.0

18.0

Control Unquenched TAML Quenched TAML

Oxygen p

roduced (

ppm

)

Trial 1

Trial 2

ANOVA or analysis of variance compares between and within the means of the groups to determine significance

ANOVA results

P-value=1.64 x 10^-4

Implies a confidence level of 99.99% that the results varied significantly

ANOVA Test

Dunnett’s test

• Compares experimental groups back to the control group. • If T value > T-Crit value than the results were

significant. T-Crit =3.03

T value T-Crit Conclusion

Quenched TAML 6.241 3.03 Significant

Unquenched TAML

6.645 3.03 Significant

Interpretation-Both quenched and un quenched TAML had a significant effect

P-value-0.212274

P-value cutoff-0.05

Conclusion-not significant

Interpretation- Quenched TAML did not have a significantly different effect than unquenched TAML

Variance between experimental groups

Reject the null hypothesis and accept the alternative that TAML improved algal productivity by reducing pollution levels

The results indicate that there may be high amounts of pollution present in panther hollow pond

TAML does not appear to harm algal growth, quenched or unquenched

Conclusions

Test was performed during winter when Algal levels were low to below freezing temperatures ◦ May have caused inaccurate results by having

strangely low amounts of algae

Water samples were taken from the mouth of the stream that feeds the pond where the water level allowed for the bottle to be filled. ◦ May not be a correct indication of algae levels in the

pond

Limitations

Mr. Mark Krotec, PTEI

Dr. Terry Collins, Thomas Lord Professor of Chemistry at Carnegie Mellon University

http://www.chem.cmu.edu/groups/Collins/about/about.html

“Little Green Molecules” by Terrence Collins and Chip Walter

References

Control Unquenched

TAML

Quenched

TAML

Trial 1 7.8 ppm 16.7 ppm 15.9 ppm

Trial 2 8.2 ppm 16.1 ppm 15.8 ppm

Average 8.0 ppm 16.4 ppm 15.85 ppm

Data

First ANOVA

SUMMARY

Groups Count Sum Average Variance

Column 1 2 16 8 0.08

Column 2 2 32.8 16.4 0.18

Column 3 2 31.7 15.85 0.005

ANOVA

Source of Variation SS df MS F P-value F crit

Between Groups 88.32333 2 44.16167 499.9434 0.000164 9.552094

Within Groups 0.265 3 0.088333

Total 88.58833 5

Variance between two experimental

groups

Anova: Single Factor

SUMMARY

Groups Count Sum Average Variance

Column 1 2 32.8 16.4 0.18

Column 2 2 31.7 15.85 0.005

ANOVA

Source of Variation SS df MS F P-value F crit

Between Groups 0.3025 1 0.3025 3.27027 0.212274 18.51282

Within Groups 0.185 2 0.0925

Total 0.4875 3