Bio Lab
Daphny Maldonado
Bio Lab 2107
Kiah Britton
W 10-12:30
Is H20 Bad for You?
Abstract:
In the village of Gopher Hollow there’s a cluster of Blue Baby Syndrome. There were four infants affected by this cluster. The families from the infants would collect their water from wells. We have to determine what’s the source of the high levels of nitrites in the water. The four sources that could be the point of contamination are a new subdivision, textile plant, an organic farm, and a mountain lake. We had to find the concentration of each known standard and unknown standard. We did this by using a spectrophotometer. The results were the following, the organic farm with a herd of 50 cows and a 10 acre field of zucchini had the highest levels …show more content…
of nitrites.
Introduction:
Blue Baby Syndrome is a condition that affects many infants. This condition makes the baby’s skin turn blue because of the lack of oxygen. This condition can exhibit lethargy, vomiting and not being able to breathe. It can even lead to death in rare cases.
This condition is caused by the excess amount of nitrate that is then converted into nitrite by the digestive system. The hemoglobin then reacts with the nitrites to form
Methemoglobin. Methemoglobin is not a problem in adults since they have an enzyme that converts methemoglobin back to hemoglobin. Infants don’t have many of the enzyme to convert methemoglobin to hemoglobin, resulting in Blue Baby Syndrome. For
example in Gopher Hollow there is a cluster of Blue Baby Syndrome. Four infants have been affected. The families of these infants all collect their water from a community well.
Nitrates and Nitrites can be found in many things. For example in vegetables, like carrots, lettuce and spinach. We do get many of our nitrogen from the food we eat but also through the water we drink.(1) Levels above 10 ppm of Nitrate/ Nitrites are not dangerous but accumulations of it in soil and groundwater can cause high levels of these compounds.(1) Causing high level of nitrites in drinking water and especially in wells. If a mountain lake that has become overgrown with algae then there could be an increase of Blue Baby Syndrome cases. The lake could be a main water source.
Methods:
The first step in determining the culprit location is to create a graph that represents the relationship between the amounts of nitrites in the solution and the optical density of standards of known concentrations. These standards are the control group. Then a spectrophotometer was turned on was allowed to warm up for 15 minutes. There were nine standards to be tested using the spectrophotometer. One ml of each standard was labeled and placed in 13 by 85mm glass test tubes. The concentrations were: distilled water, 0.5ppm, 1.0ppm, 2.0ppm, 4.0ppm, 6.0 ppm, 10.0 ppm, 20.0 ppm, and 40.0 ppm.
Using the P20 micropipettor, 20 microliters of the color indicator reagent ( 1naphthylethylenediamine sulfanilamide) was added to each standard sample and mixed throughly. A new micropipette tip was used to prevent mixing of chemicals. The samples were left out to sit for five minutes to insure full color development. Then the spectrophotometer’s wavelength was set to 550nm because the nitrite indicator absorbs light at 55onm(1). I ml of the distilled water sample was transferred into a clean cuvette
using a plastic transfer pipette. The cuvette was then placed inside the spectrophotometer . The button labeled “blank” was pressed and the OD reading was
0.000A . After getting the OD, the blank sample was placed back into its original test tube. Then 1 ml of the 0.5ppm standard was transferred into a clean cuvette and placed inside the spectrophotometer for OD measurement. Once the measurement appears on the screen record it. This step was repeated for the remaining standard samples. After measuring the OD each sample was transferred back into its original test tube. With the data collected, a graph was plotted with the Optical Densities on the vertical axis and the nitrite concentrations on the horizontal axis. Once the graph was plotted, a standard curve and the line of best fit were created and put on the graph. Whenver the OD reading was above 1.0 it’s omitted from the graph because it means that there very light being detected. To get the line of best fit the equation y=mx+b was used and m=slope and b= y intercept. Then the unknown samples of the sites thought to be the cause of the Blue baby syndrome were tested using the same techniques as to find the standard samples. The first step, 2 ml of each water sample were place into four 13 by 85 mm glass test tubes. Then using a P200 micropipettor, 40 microliters of sulfanilamide was added to each test tube. The test tubes were allowed five minutes to develop color.
Then 1 ml of each sample was transferred into a clean cuvette and the optical density was measured using the spectrophotometer. By using the previously made standard curve , the concentration of the nitrites in the unknown sites can be found.
Results:
In the first part of the experiment the concentrations for the known standards were:
0 for distilled water, .071M for .5ppm, .134M for 1ppm, .251M for 2ppm, .040M for
4ppm, .597M for 6ppm, .940M for 10ppm, 1.107M for 20ppm, and 2.060M for 40ppm.
(Figure 1) The concentrations for the four samples were as follows: .293M for A, .555M for B, .954M for C and .617M for D. (Figure 2) To find which sample had the highest levels of nitrites use the line of best fit to solve for concentration. The slope was found to be 0.1349. To find the concentrations levels of nitrites use the optical densities found already and divide by the slope which was 0.1349. For sample A the nitrite concentration was 2.17ppm and sample b was 4.11ppm. Then sample c the concentration levels for nitrite was 7.07ppm and sample d was 4.57 ppm. (Figure 3)
Figure 1:
The concentration for the known standards.
Concentration
OD
Distiiled water
0.000
0.5 ppm
0.071
1.0 ppm
0.134
2.0 ppm
0.251
4.0 ppm
0.040
6.0 ppm
0.597
10.0 ppm
0.940
20.0 ppm
1.107
40.0 ppm
2.060
Figure 2:
The concentration for the unknown standards.
Unknown samples
OD
Distilled Water
0.000
A
0.293
B
0.555
C
0.954
D
0.617
Figure 3: The final concentration levels of nitrites for the sample sites.
Site
Concentrations
A
2.17
B
4.11
C
7.07
D
4.57
Discussion:
Based off the results that we got from the experiment the main source for high levels of nitrites in the water was sample C. Which was the organic farm with a herd of
50 cows and a 10 acre field of zucchini. The hypothesis did not support the final result.
The highest level of nitrites was 7.07 ppm which was accurately found using the best fit line.(Figure 4) From this result it was disproven that the mountain lake would be the cause of high levels of nitrites in the water. The results supported the cause of Blue baby syndrome and how the organic farm was the main source of the contamination of the water. Which caused the high levels of nitrites and causing the cluster of blue baby syndrome in the village of Gopher Hollow.
Citations: gutzler, stephanie , and mathhew brewer . principles of biology . plymouth : hayden mcneil , 2013.
Print.
Bio Lab 2107
Kiah Britton
W 10-12:30
Is H20 Bad for You?
Abstract:
In the village of Gopher Hollow there’s a cluster of Blue Baby Syndrome. There were four infants affected by this cluster. The families from the infants would collect their water from wells. We have to determine what’s the source of the high levels of nitrites in the water. The four sources that could be the point of contamination are a new subdivision, textile plant, an organic farm, and a mountain lake. We had to find the concentration of each known standard and unknown standard. We did this by using a spectrophotometer. The results were the following, the organic farm with a herd of 50 cows and a 10 acre field of zucchini had the highest levels …show more content…
of nitrites.
Introduction:
Blue Baby Syndrome is a condition that affects many infants. This condition makes the baby’s skin turn blue because of the lack of oxygen. This condition can exhibit lethargy, vomiting and not being able to breathe. It can even lead to death in rare cases.
This condition is caused by the excess amount of nitrate that is then converted into nitrite by the digestive system. The hemoglobin then reacts with the nitrites to form
Methemoglobin. Methemoglobin is not a problem in adults since they have an enzyme that converts methemoglobin back to hemoglobin. Infants don’t have many of the enzyme to convert methemoglobin to hemoglobin, resulting in Blue Baby Syndrome. For
example in Gopher Hollow there is a cluster of Blue Baby Syndrome. Four infants have been affected. The families of these infants all collect their water from a community well.
Nitrates and Nitrites can be found in many things. For example in vegetables, like carrots, lettuce and spinach. We do get many of our nitrogen from the food we eat but also through the water we drink.(1) Levels above 10 ppm of Nitrate/ Nitrites are not dangerous but accumulations of it in soil and groundwater can cause high levels of these compounds.(1) Causing high level of nitrites in drinking water and especially in wells. If a mountain lake that has become overgrown with algae then there could be an increase of Blue Baby Syndrome cases. The lake could be a main water source.
Methods:
The first step in determining the culprit location is to create a graph that represents the relationship between the amounts of nitrites in the solution and the optical density of standards of known concentrations. These standards are the control group. Then a spectrophotometer was turned on was allowed to warm up for 15 minutes. There were nine standards to be tested using the spectrophotometer. One ml of each standard was labeled and placed in 13 by 85mm glass test tubes. The concentrations were: distilled water, 0.5ppm, 1.0ppm, 2.0ppm, 4.0ppm, 6.0 ppm, 10.0 ppm, 20.0 ppm, and 40.0 ppm.
Using the P20 micropipettor, 20 microliters of the color indicator reagent ( 1naphthylethylenediamine sulfanilamide) was added to each standard sample and mixed throughly. A new micropipette tip was used to prevent mixing of chemicals. The samples were left out to sit for five minutes to insure full color development. Then the spectrophotometer’s wavelength was set to 550nm because the nitrite indicator absorbs light at 55onm(1). I ml of the distilled water sample was transferred into a clean cuvette
using a plastic transfer pipette. The cuvette was then placed inside the spectrophotometer . The button labeled “blank” was pressed and the OD reading was
0.000A . After getting the OD, the blank sample was placed back into its original test tube. Then 1 ml of the 0.5ppm standard was transferred into a clean cuvette and placed inside the spectrophotometer for OD measurement. Once the measurement appears on the screen record it. This step was repeated for the remaining standard samples. After measuring the OD each sample was transferred back into its original test tube. With the data collected, a graph was plotted with the Optical Densities on the vertical axis and the nitrite concentrations on the horizontal axis. Once the graph was plotted, a standard curve and the line of best fit were created and put on the graph. Whenver the OD reading was above 1.0 it’s omitted from the graph because it means that there very light being detected. To get the line of best fit the equation y=mx+b was used and m=slope and b= y intercept. Then the unknown samples of the sites thought to be the cause of the Blue baby syndrome were tested using the same techniques as to find the standard samples. The first step, 2 ml of each water sample were place into four 13 by 85 mm glass test tubes. Then using a P200 micropipettor, 40 microliters of sulfanilamide was added to each test tube. The test tubes were allowed five minutes to develop color.
Then 1 ml of each sample was transferred into a clean cuvette and the optical density was measured using the spectrophotometer. By using the previously made standard curve , the concentration of the nitrites in the unknown sites can be found.
Results:
In the first part of the experiment the concentrations for the known standards were:
0 for distilled water, .071M for .5ppm, .134M for 1ppm, .251M for 2ppm, .040M for
4ppm, .597M for 6ppm, .940M for 10ppm, 1.107M for 20ppm, and 2.060M for 40ppm.
(Figure 1) The concentrations for the four samples were as follows: .293M for A, .555M for B, .954M for C and .617M for D. (Figure 2) To find which sample had the highest levels of nitrites use the line of best fit to solve for concentration. The slope was found to be 0.1349. To find the concentrations levels of nitrites use the optical densities found already and divide by the slope which was 0.1349. For sample A the nitrite concentration was 2.17ppm and sample b was 4.11ppm. Then sample c the concentration levels for nitrite was 7.07ppm and sample d was 4.57 ppm. (Figure 3)
Figure 1:
The concentration for the known standards.
Concentration
OD
Distiiled water
0.000
0.5 ppm
0.071
1.0 ppm
0.134
2.0 ppm
0.251
4.0 ppm
0.040
6.0 ppm
0.597
10.0 ppm
0.940
20.0 ppm
1.107
40.0 ppm
2.060
Figure 2:
The concentration for the unknown standards.
Unknown samples
OD
Distilled Water
0.000
A
0.293
B
0.555
C
0.954
D
0.617
Figure 3: The final concentration levels of nitrites for the sample sites.
Site
Concentrations
A
2.17
B
4.11
C
7.07
D
4.57
Discussion:
Based off the results that we got from the experiment the main source for high levels of nitrites in the water was sample C. Which was the organic farm with a herd of
50 cows and a 10 acre field of zucchini. The hypothesis did not support the final result.
The highest level of nitrites was 7.07 ppm which was accurately found using the best fit line.(Figure 4) From this result it was disproven that the mountain lake would be the cause of high levels of nitrites in the water. The results supported the cause of Blue baby syndrome and how the organic farm was the main source of the contamination of the water. Which caused the high levels of nitrites and causing the cluster of blue baby syndrome in the village of Gopher Hollow.
Citations: gutzler, stephanie , and mathhew brewer . principles of biology . plymouth : hayden mcneil , 2013.
Print.