The Contamination of Phosphorus
in 4 Different Locations
Jess Withey
Research Advisor: Dr. Carl Binz
Secondary Advisor: Dr. Edward Maslowski
May 2013
Senior Biochemistry Thesis
Objectives
The main objectives for the presented research were:
To analyze total phosphorus concentration using an
absorbic acid method of spectrometry
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Reflects the level of phosphorus in water
Measure of both soluble and non-soluble organic phosphorus
To compare the concentrations of phosphorus in four
different locations that relate to the Mississippi River
including:
• Pike Lake Chain; Park Falls, Wisconsin
• Flambeau River; Park Falls, Wisconsin
• Mississippi River; Dubuque, Iowa
• Mississippi River; Clinton, Iowa
Background
• Form phosphates (PO4-3)
• Exist in 3 forms:
▫ Orthophosphate-produced by natural processes and
are found in sewage
▫ Metaphosphate-treat boiler waters and are in
detergents
▫ Organically bound phosphate-important in nature;
result from the breakdown of organic pesticides
 May exist as loose fragments in solution or in bodies of
aquatic organisms
• Occur in minerals and foods
• Found in oceans—very low at surface and increases
as depth increases due to organic debris
• Found in rivers and lakes—results in generating
excessive algae growth
Pollutant Sources of Phosphorus
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Wastewater and Septic System Effluent
Detergents
Fertilizers, Leaves, Grass Clippings
Animal Waste
Development/Paved Surfaces
Industrial Discharge
Forest Fires
Phosphorus Health Effects
• Phosphorus is a dietary requirement with the
recommended intake being 700mg/day (normal
diet provides 1000-2000mg/day)
• Phosphorus exists as phosphate.
• Considered as the second most profuse/abundant
mineral in the human body.
• Essential for activities for the brain, kidneys, heart,
and blood
Occur in Minerals & Foods
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Eggs
Meats
Poultry
Fish
Nuts
Dairy Products
Sunflower Seeds
Rice
White Bread
Potatoes
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Broccoli
Peas
Peanut Butter
Tuna
Pork
Functions
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Growth and Development
Bone Formation
Digestion
Excretion
Weakness
Brain Function
Protein Formation
Hormone Balance
Energy Extraction
Cell Repair
Chemical Reactions/Production of Energy
Excessive Intake in Humans
• Diarrhea
• Calcification in organs and soft tissue (kidneys)
• Prevent absorption of iron, calcium, magnesium,
and zinc
• Reduces Vitamin D production and blood calcium
• Kidney Damage
• Osteoporosis
Environmental Intake
• Needed by plants to produce energy and for their
reproductive systems
• Essential for the successful early establishment
of vegetables, flowers, and lawn grasses.
The Phosphorus Cycle
Phosphorus Environmental Effects
• Algae growth is limited by the available supply of
phosphorus, so large amounts can trigger large
quantity growth
• When algae die, they are decomposed by
bacteria, which use dissolved oxygen.
• Oxygen concentrations can drop too low leaving
fish and other aquatic organisms with the
inability to breath, resulting in death.
Eutrophilication
• Process that results from accumulation of
nutrients in lakes or other water bodies
• “The aging of lakes”
• Natural process that takes thousands of years
• Can be greatly accelerated by human activities
that increase the rate at which nutrients enter
the water
Reducing Phosphorus Contaminants
• Effective nutrient management
▫ Keep leaves, tree seeds and lawn clippings out of
gutters, streets, and ditches
▫ Pick up pet waste
▫ Control soil erosion
• Phosphorus information regarding:
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Fertilizer recommendations
Phosphorus index
Research projects
Water quality impact
My Interests
Pike Lake
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806 acres
Mean depth=11ft
Max depth=17ft
pH=6.2-6.7
Water brown in color
▫ Tamrick tree swamps
▫ Inflowing streams from
wetland drainage
• Formed approximately
10,000 years ago
during last glacial
retreat from WI.
• High Drainage areas to Pike Lake:
▫ Direct watershed: land area that drains to the lake
by runoff
▫ Contributing watershed: land areas that drain to
the lake by way of a defined channel/stream
▫ Total watershed: the direct drainage watershed
area plus the contributing watershed area
• Drainage areas dominated by forests and
wetlands, as well as 3 other lakes connected and
their creeks.
• All drainage areas contribute to the chain’s large
phosphate concentration.
• Bottom of lake contains
some of the highest
available phosphorous of
any soil in WI.
▫ Stream inputs
▫ Boat traffic
▫ Sediment resuspension
• Phosphorous generates
algae blooms and weeds.
• Nitrates present due to
septic systems.
• Iron and sulfur present from
rocks.
Hypothesis
• The water samples will decrease in molarity
starting from Northern Wisconsin (Pike Lake) to
the southern part of the Mississippi River in
Clinton, Iowa.
Methods/Procedure
• Testing strips can detect phosphate ions in
water.
▫ Didn’t go low enough
in concentration
Methods/Procedure
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Water samples containing phosphorus were collected
once a month from December to April 2013.
The standard stock solution was prepared from sodium
phosphate (1.00x10-3M) to prepare a calibration curve to
help analyze the collected samples.
The stock solutions were then diluted to the predicted
range of the sample solutions:
(1.00x10-4M, 5.5x10-5M,1.00x10-6M)
1mL of ammonium molybdate was added to 5mL of the
water sample.
0.7mL of aminonaphtholsulfonic acid reagent was added.
Samples were left to sit for 30 minutes with continuous
swirling.
Absorbance values were measured at 690nm using an
OceanOptics UV Vis Spectrometer.
• The three standards were plotted to determine
the experiment’s calibration curve.
• Beers Lamberts Law was applied to analyze and
determine the unknown concentrations.
Beers Law
A=εbc
“ε " is the molar absorptivity (in units of per
molar per cm, M-1 cm-1)
"b" is the path length (in units of centimeters,
cm),
"c" is the concentration (in units of morality , M).
Figure 1. represents an example of a water sample test from the standard stock solution and the
standards that were prepared to determine the experiment’s calibration curve. The unknown
concentrations of the water samples were then derived using the absorbance and concentration curve,
as shown above.
Molarity of Samples
Trial
Sample (10-5M)
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Pike Lake
2.93
2.85
3.03
2.88
2.87
Flambeau River
1.63
1.88
2.03
1.93
1.90
MI River-Dubuque
1.25
1.24
1.21
1.16
1.25
MI River-Clinton
.733
.851
.912
.766
.854
The exact concentration of total phosphorus in
the samples were determined and shown
below:
Area of Samples Concentration Ranges:
• Pike Lake, Park Falls, WI
2.85x10-5-3.03x10-5M
• Flambeau River, Park Falls, WI
1.63x10-5-2.03x10-5M
• Mississippi River, Dubuque, IA
1.16x10-5-1.25x10-5M
• Mississippi River, Clinton, IA
7.33x10-6-9.12x10-6M
Phosphorus Concentration Over Time
3.5
Concentration (10-5M)
3
2.5
Pike Lake
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Flambeau
Dubuque
1.5
Clinton
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0.5
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Sample Trial
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Conclusions
• Pike Lake had the highest phosphorus
concentration of all the sites.
• Based on this study, it can be concluded that
the amount of phosphorus in the water
decreases as it goes further south.
Experimental Outcomes
• An understanding of how to determine the
concentration of phosphorous using UV Vis
spectroscopy
• The technique to analyzing samples
• The concentrations of phosphorus in the
samples were calculated using Beers Law
Future Outlooks
• Test the water during different seasons
▫ Difference in algae blooms
• Do testing at different depths in the water
▫ Surface vs. Bottom
• Test soil in water sources for phosphorous
Acknowledgements
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Loras College
Dr. Carl Binz
Dr. Edward Maslowsky
Professors
Family and Friends
Works Cited
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http://www.lenntech.com/periodic/elements/p.htm
http://www.lenntech.com/periodic/elements/n.htm
http://water.epa.gov/type/rsl/monitoring/vms56.cfm
http://sd.lake-link.com/Wisconsin-LakeFinder/lake.cfm/3955/Pike-Lake-Price-County-Wisconsin
http://dnr.wi.gov/lakes/grants/Project.aspx?project=10100227
http://linus.chem.ku.edu/hewlett/Chem188/PhosphateLaboratory
/procedures%20for%20phosphate%20determination.htm
http://bcn.boulder.co.us/basin/data/NEW/info/TP.html
http://edis.ifas.ufl.edu/ss490
http://www.water-research.net/Watershed/phosphates.htm
http://www.indigo.com/test_strips/water_testing/33817Pphosphate-levels-test-strips-0-100ppm.html
http://www.mda.state.mn.us/news/publications/chemfert/reports
/phosphorusguide.pdf
http://www.organicfacts.net/health-benefits/minerals/healthbenefits-of-phosphorus.html
http://www.livestrong.com/phosphorus/