Advanced Placement Biology
®
292
AP Biology
Lab 12
EDVO-Kit #
Dissolved Oxygen &
Aquatic Primary Productivity
Storage:
Store entire experiment at room temperature
EXPERIMENT OBJECTIVE
The objective of this experiment is to understand:
1) the factors that affect the solubility of dissolved
gases in aquatic environments, 2) the effects of
light and nutrients on photosynthesis, and 3) the
relationship between dissolved oxygen, photosynthesis and respiration. Students should be able to
describe a method to measure dissolved oxygen,
and define primary productivity and factors which
influence it.
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EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
AP
Biology
Table of Contents
Lab #
12
Page
Experiment Components
3
Experiment Requirements
Background Information
3
4
Experiment Procedures
Experiment Overview
Part A. The Winkler Method for Dissolved Oxygen
7
8
Part B. Primary Productivity
Laboratory Extensions
13
20
Study Questions
27
Instructor's Guidelines
Notes to the Instructor
Pre-Lab Preparations
Study Questions and Answers
Material Safety Data Sheets
Advanced Placement (AP) Program is a
registered trademark of the College Entrance
Examination Board. These laboratory materials
have been prepared by EDVOTEK, Inc. which
bears sole responsibility for their contents.
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29
30
33
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EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
3
AP
Biology
Experiment Components
This experiment
is designed for
10 lab groups
A
B
C
D
E
F
G
Sodium iodide
Sodium hydroxide
Manganese sulfate
Starch solution
Sodium thiosulfate
Nitrogen enrichment solution, concentrate
Phosphorous enrichment reagents
Pipets
Lab #
Storage:
Store entire
experiment at
room temperature.
Requirements
All components are
intended for
educational research
only. They are not to
be used for
diagnostic or drug
purposes, nor
administered to or
consumed by
humans or animals.
EDVOTEK, The
Biotechnology
Education Company,
and InstaStain are
registered
trademarks of
EDVOTEK, Inc.
•
•
•
•
•
•
•
•
•
•
•
•
•
•
Plastic window screens
Sealable containers or BOD bottles, 50
Grow lights
Pond/lake water
Goggles
Sulfuric acid, concentrated
Glass pipets
Burettes
Flasks
Beakers
Gloves
Filter paper
Linear graph paper
Aluminum foil
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4
EDVO-Kit # 292
AP
Biology
Lab #
Background Information
12
Dissolved Oxygen & Aquatic Primary Productivity
Dissolved Oxygen & Aquatic Primary Productivity
Oxygen plays a major role in the biochemical reactions used by
organisms to produce energy for life. During oxidative phosphorylation
and electron transport, it is generally the final electron acceptor. The
greatest number of ATP’s are produced per mole of glucose when
oxygen is available. Its concentration in the environment can limit the
maximal rates of metabolism. In air, there is ample concentration of
oxygen available for utilization by living organisms. It represents nearly
20% of the total gases in the air. There is approximately 200ml of O2 per
liter of air, which is 9 mM O2 /L. The situation in aquatic environments is
quite different.
In aquatic environments, oxygen is not very soluble and its solubility is
directly affected by the concentration of dissolved ions such as salt,
the pH, and the temperature. Salinity is the content of dissolved salts in
water. It is usually expressed in parts per thousand (ppt). The solubility
of O2 is inversely proportional to the concentration of salt and temperature. As the salt concentration and temperature increase, the
solubility of O2 decreases. Even at its maximum solubility, which is in
fresh water at 0°C, its concentration is only 25% of the amount in air. In
practical situations, the concentration of O2 in aquatic environments
usually does not exceed 3-4% of the concentration of O2 in air. This is
only 6.0ml O2/L or 0.3 mM O2 /L. In salt water, O2 concentration would
be less since oxygen is less soluble in solutions of increasing ionic
strength.
Since the maximum concentration of dissolved O2 in water is dependent upon other components dissolved in water, O2 level is often used
in water quality and pollution testing. The higher the level of dissolved
oxygen (DO), the better the water quality. Because of the low concentration of O2 in aquatic environments, it has been suggested that
O2 is a major limiting nutrient for life. It is interesting to note that the
largest aquatic animals, the whales, are air breathing mammals.
In addition to the chemical factors that influence dissolved oxygen,
biological processes such as photosynthesis and respiration affect the
maximum amount of aqueous oxygen concentrations. There are both
biological demands and chemical demands placed on the oxygen
available in water. These are often called the Biological Oxygen
Demand (BOD), and the Chemical Oxygen Demand (COD). Photosynthesis in aquatic environments performed by plants, and small phytoplankton which are one-celled plants, will increase the concentration of DO in the water. Respiration will decrease the concentration of
DO in an aquatic environment. Therefore during the day, when
photosynthesis is at a maximum, the concentration of DO will increase.
During the night, the concentration of DO will decrease.
Ecologists will often study an ecosystem by measuring or estimating the
“primary productivity”. Energy to support the life requirements of
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EDVO-Kit # 292
5
Dissolved Oxygen & Aquatic Primary Productivity
Dissolved Oxygen & Aquatic Primary Productivity
animals, plants, and bacteria generally enters an ecosystem as light energy,
which is converted by photosynthetic organisms to chemical energy.
Primary productivity is defined as the rate at which plants assimilate the
energy of sunlight. This rate directly affects the growth of plants and other
chlorophyll containing organisms. This in turn affects the growth of animals
that feed on plants and others which are at higher trophic levels. Primary
productivity therefore is at the base of the trophic structure ( the organization of feeding relationships) in an ecosystem.
AP
Biology
Lab #
12
The equation for primary productivity, which is the utilization of carbon
dioxide and water to produce glucose and oxygen during photosynthesis is;
6 CO2 + 6 H2O
→
C6 H12 O6 + 6 O2
The accumulation of organic matter through the growth and reproduction
of the plant is not the only productivity possible. A significant portion of the
sugar produced by photosynthesis is used by the plant for other synthetic
and maintenance reactions during respiration. Therefore, the gross primary
productivity (GPP) is the sum of the organic material produced plus the
respiration rate (Rs) of the plant or GPP = NPP + Rs. Net primary productivity
(NPP) equals the gross productivity minus the respiration rate (Rs) or
NPP = GPP – Rs. Net productivity is therefore a measurement of growth and
reproduction. Gross productivity is a measurement of growth and reproduction plus the respiration rate.
In terrestrial environments, plant primary productivity is usually measured in
terms of the increase in the amount of plant Biomass. In aquatic environments, we measure the plant primary productivity by gas exchange, since
the concentration of dissolved oxygen in water is easily determined. The
classical method for determining the productivity of an aquatic environment
is the light and dark bottle procedure. In a natural setting, for example,
sealed bottles containing samples of pond water would be suspended at
different depths beneath the surface of the pond. A clear “light” bottle
which allows sunlight to enter, and a “dark” bottle which excludes sunlight,
would be suspended at each depth. In the laboratory, one can simulate
the attenuation of sunlight, which increases as depth increases, by wrapping
the light bottles with screens. We will assume that the respiration is equal at
all depths. In the light bottles, the production of oxygen by photosynthesis
and the consumption of oxygen occur simultaneously. Therefore the
change in oxygen levels in the light bottles is a direct measure of the “net
productivity”. In the dark bottles, oxygen is consumed since only respiration
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Background Information
From the above equation, the amount of oxygen produced can be directly
related to the amount of carbon dioxide consumed. For each mole of O2
produced, one mole of CO2 is consumed. One milliliter of O2 gas contains
1.432 mg of oxygen. Therefore, based on formula weights, one can calculate that for each milliliter of oxygen produced, approximately 0.536 mg of
carbon has been consumed.
6
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
AP
Biology
Dissolved Oxygen & Aquatic Primary Productivity
Lab #
occurs. Therefore, the change in oxygen levels in the dark bottles is a
measure of respiration (Figure 1). Addition of the amount of oxygen
consumed by respiration in the dark bottle and the oxygen produced
in the light bottle, yields gross productivity. The respiration rate includes
that of plants, animals, and bacteria. Therefore, only the gross productivity measurement is totally valid.
12
Figure 1:
Gross Productivity
Determined by Light/Dark
Bottle Method
The level of dissolved oxygen (DO) in water is a direct measurement of
the quality of the water. This is affected by physical conditions such as
temperature, salinity, and pH. In addition, it is dramatically affected by
both the Biological Oxygen Demand (BOD), and Chemical Oxygen
Demand (COD). Lakes loaded with decaying matter have low levels
of oxygen, and a high BOD, since the oxygen is being consumed by
bacteria and algae. This exercise demonstrates the methodology
used for measuring dissolved oxygen and determining the primary
productivity of a natural body of water.
( L )ight Bottle
Net
Productivity
DO2 (ml O2/L)
Background Information
The Winkler method is used to determine dissolved oxygen. This
procedure is an iodometric method. The iodide ion is an effective
reducing agent which has been widely used for the quantitative
analysis of many oxidants. Generally, sodium thiosulfate is used to
titrate the iodine liberated by the chemical reaction. The endpoint is
determined by the loss of color during titration. This procedure is
explained in detail under Part A, Student Experimental Procedures.
( I )nitial Bottle
Gross
Productivity
Respriation
( D )ark Bottle
0
24
L - I = Net Productivity
I - D = Respiration
L - D = Gross Productivity
Incubation Time (Hours)
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EDVO-Kit # 292
7
Dissolved Oxygen & Aquatic Primary Productivity
AP
Experiment Overview
Biology
EXPERIMENT OBJECTIVES:
Students will:
Understand the factors that affect the solubility of dissolved gases in
aquatic environments
2.
Understand the effects of light intensity and nutrients on the rate of
primary productivity
3.
Understand the relationship between dissolved oxygen and the
processes of photosynthesis and respiration and the affect on primary
productivity
4.
Be able to describe a method to measure dissolved oxygen, and
define primary productivity and factors which influence it .
WORKING HYPOTHESIS
If light intensity and the addition of nutrients affect the rate of photosynthesis in aquatic environments, then primary productivity will also be
affected.
LABORATORY SAFETY
Gloves and safety goggles should be worn routinely as good laboratory
practice.
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Experiment Procedure
1.
Lab #
8
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
AP
Biology
Lab #
Experiment Procedure
12
Part A: The Winkler Method for Dissolved Oxygen
THE WINKLER METHOD FOR DISSOLVED OXYGEN
The Winkler method will be used to determine the concentration of
dissolved oxygen in water samples. The samples are first treated with
an excess of manganese sulfate, sodium iodide, and sodium hydroxide. The white manganese (II) hydroxide precipitate that forms will
rapidly react with the dissolved oxygen in the samples to form manganese (III) hydroxide. When the samples are acidified by the addition of
sulfuric acid, the manganese (III) oxidizes iodide to iodine. The concentration of the liberated iodine, I2, is titrated with sodium thiosulfate
in the presence of a starch indicator solution.
It is important to note that the starch is added after the bulk of the
iodine has been reduced. In acidic solutions, the starch would be
decomposed by a large excess of iodine. For fine analysis, the starch
is added after the titration has begun, near the point when the
solution has become a faint, pale yellow color. For purposes of this
laboratory, the starch can be added at the beginning of titration to
facilitate end point detection.
The balanced equations for the series of chemical reactions used in
the Winkler method are shown below.
4 Mn (OH)2 (s) + O2 + 2 H2O →
4 Mn (OH)3 (s)
When acidified by addition of sulfuric acid;
2 Mn (OH)3 (s) + 2 I- + 6 H+
→
I2 + 6 H2O + 2 Mn2+
The liberated iodine is titrated to endpoint with sodium thiosulfate as
shown below.
2 (S2 O3)-2 + I2
→
S4 O6-2 + 2I-
When the titration is nearly complete, the solution will begin to change
from a purple/brown color to a colorless solution. As can be seen from
the above reactions, four moles of sodium thiosulfate titrant, Na2S2O3,
are required per mole of dissolved oxygen, O2. The concentration of
the sodium thiosulfate titrant has been adjusted so one milliliter of
sodium thiosulfate solution equals one milligram of dissolved oxygen
per liter of sample, 1 mg DO/L.
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EDVO-Kit # 292
9
Dissolved Oxygen & Aquatic Primary Productivity
AP
Part A: The Winkler Method for Dissolved Oxygen
Forming the Manganese (III) Hydroxide Precipitate
1.
Obtain a water sample from the instructor. Samples should be at
either 5°, 20° or 30°C. Record the temperature of the water with a
thermometer. Different groups will use water at different temperatures.
2.
Place the water sample into your 250-300ml “BOD” (Biological Oxygen
Demand) bottle. Any container which can be closed with an airtight
seal is appropriate.
Allow the sample to overflow the container so that it will be
completely filled.
•
Close the container. Turn the container upside down and use a
paper towel to remove any water which is around the outside of
the stopper or lid.
Create manganese hydrogen precipitate in your water sample:
•
Open the container and carefully pipet 2ml of manganese sulfate
into the container. Make sure the pipet tip is below the water
surface in the container.
•
With a fresh pipet, add 2ml of the NaOH/NaI (alkaline-iodide)
solution into the sample in the container. Make sure the pipet tip
is below the surface of the water in the container.
•
Stopper or seal your container. Carefully invert the bottle to allow
for complete mixing of the sample, manganese sulfate, and
NaOH/NaI.
•
This precipitate is critical to the determination of dissolved oxygen. If a precipitate fails to form, repeat procedure.
•
Allow the precipitate to settle for 10-15 minutes or until approximately 50% of the volume in the container is occupied by the
precipitate.
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Lab #
12
Experiment Procedure
3.
•
Biology
10
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Part A: The Winkler Method for Dissolved Oxygen
Lab #
Setting up the Burette
12
4.
While waiting for the manganese hydroxide precipitate to settle,
set up your burette.
•
•
5.
Test the proper functioning and seal of the petcock at the bottom
of the burette.
•
Experiment Procedure
•
•
6.
Label a beaker ‘Burette Waste Solution’ and place it under the
tip of the burette.
Close the petcock and fill the burette with distilled water.
Open and close the petcock. When open, the water should
flow out. When closed, the water should not leak out of the
burette tip or around the petcock assembly. Ask your instructor for help if the burette leaks. Practice slightly opening the
petcock so that the water just drips out slowly.
Completely drain the burette. Close the petcock.
•
•
•
7.
Attach a burette to a ring stand with a clamp.
Adjust so it is vertical and the volume gradations are easily
visible. Allow enough space under the burette for a 500ml
flask.
To remove any residual water, fill the burette with 3ml of the
sodium thiosulfate solution.
Open the petcock and allow the sodium thiosulfate solution to
drain from the burette. Again, practice opening the petcock
so that the sodium thiosulfate solution just drips out slowly.
Completely empty the burette. Close the petcock.
After closing the petcock, completely fill the burette with the
sodium thiosulfate solution.
Addition of the Hazardous Concentrated Sulfuric Acid
8.
Go to the station set up by the instructor for pipeting the Concentrated Sulfuric Acid.
•
•
Caution!
This should be performed by the
instructor. The instructor will be
wearing gloves and goggles.
Exercise extreme caution!!
•
•
Open your container and place it on the lab bench.
Standing away from the container, have the instructor pipet
2 ml of the concentrated sulfuric acid directly into your
container. The tip of the pipet should be below the surface of
the solution in your container.
Stopper the container and mix by carefully inverting the
container several times.
Mix occasionally. You should observe that the precipitate
begins to dissolve and the sample will become yellowish as
free iodine, I2, is formed in the container. Remember that the
quantity of free I2 released in this step is directly equivalent to
the concentration of dissolved oxygen in your sample.
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EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
11
AP
Part A: The Winkler Method for Dissolved Oxygen
Titration of the Free I2
9.
Wait 5 minutes and then remove exactly 200ml of solution from the
container with a graduated cylinder and transfer the solution to a
500ml flask.
Biology
Lab #
12
10. Add 1.0ml of starch solution to the pale yellow solution in the 500ml
flask. The solution will immediately become purple. Swirl the flask
gently to completely mix.
11. Place the 500ml flask under your burette tip. Adjust the burette if
necessary. Record the initial starting volume of the sodium thiosulfate
in the burette.
12. Slowly and carefully open the petcock of the burette until the sodium
thiosulfate begins to drip out of the burette tip.
•
•
Continually swirl the 500ml flask to insure thorough mixing of the
sodium thiosulfate with the sample.
Close the petcock when the solution in the flask is a pale yellow
color.
13. Open the petcock carefully so that the sodium thiosulfate solution
drips out slowly. Continue to gently swirl the flask to insure complete
mixing.
14. Be careful when adding the titrant. When the solution starts to lighten
in color, it should only take another 2-3 drops of titrant. Try not to miss
the endpoint. The endpoint is reached when the solution has gone
from purple to completely clear. Immediately close the petcock to
stop addition of the sodium thiosulfate solution.
15. At this point, all of the free I2 has been converted to sodium iodide by
reaction with the sodium thiosulfate. The solution is colorless, since
there is no longer any I2. Record the final position of the meniscus of
sodium thiosulfate in the burette.
Record the ending volume here. ___________
Determine the total volume of sodium thiosulfate which was used to
titrate your sample to the endpoint. Calculate the difference between the starting volume recorded in Step 11 and the ending
volume recorded above.
Record the total volume of sodium thiosulfate used here in ml.
___________
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EVT 005057AM
Experiment Procedure
Record the starting burette volume here. __________
12
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Part A: The Winkler Method for Dissolved Oxygen
Lab #
16. For the sensitivity required in this laboratory experiment, the total
volume in milliliters of the sodium thiosulfate solution used to titrate
the sample to the endpoint is exactly equivalent to the dissolved
oxygen concentration in milligrams of dissolved oxygen (DO) per
liter of sample solution, mg DO/L.
12
17. Record the temperature of your sample and the amount of
dissolved oxygen in the chart. Use the nomogram of oxygen
saturation graph on this page to determine the % saturation of DO
in the sample. If your temperature lies between these values,
estimate the % saturation of DO. Record the value in Table 1.
Experiment Procedure
TABLE 1: Temperature/DO Data
Temperature Your DO mg/L
0
5
%DO Saturation Class Mean DO
(from nomogram)
mg/L
Class Mean DO Saturation
(from nomogram)
18. Determine the class
average data for the water
samples at the three
temperatures which were
available. Record the
values in Table 1.
10
15 20 25 30
Water Temperature °C
ion
rat
00
40
50
60
70
80
120
140
%
u
Sat
19. Using linear graph paper or
Graph #1 provided on
page 18, plot Both the lab
group and class means
percent saturation as
function of temperature.
1
90
30
20
10
0
2
3
4
5
6
7
8
9
10
11
12 13 14
15 16
17
Oxygen (mg per liter)
Figure 2:
Nomogram of Oxygen Saturation
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EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
13
AP
Biology
Part B: Primary Productivity
PRIMARY PRODUCTIVITY
For Part B., samples of natural water will be used to determine gross
productivity, net productivity, and respiration rates. To mimic the light
attenuation found as one goes deeper beneath the surface of a body
of water, screens will be used to reduce the light intensity. Review the
Background section of this laboratory for a complete discussion of
Primary Productivity.
Lab #
12
Day One
1.
Obtain 7 clean “BOD” (Biological Oxygen Demand) containers. Any
container which holds approximately 250-300ml and which can be
closed with an airtight seal is appropriate.
•
2.
Completely fill each bottle with water sample from the lake or
pond. Allow the sample to overflow the container so that it will
be completely filled.
Stopper or close the container. Turn the container upside down
and use a paper towel to remove any water which is around the
outside of the stopper or lid.
Label the 7 bottles as follows:
#1 - Initial
#2 - Dark
#3 - 100%
#4 - 65%
#5 - 25%
#6 - 10%
#7 - 5%
3.
Bottle #1 is the initial starting bottle and serves as a baseline.
4.
Wrap Bottle #2 in aluminum foil, for it serves as the dark (no light)
control. Place in a dark place. No photosynthesis, only respiration,
will occur here.
5.
Bottles #3 –#7 will simulate the depth in a body of water that natural
light will attenuate This is done by wrapping each bottle with a
different number of screens. Bottle #3 will have no screens with 100%
attenuation of light, Bottle #4 has 1 screen with 65% attenuation,
Bottle #5 has 3 screens with 25% attenuation, Bottle #6 has 5 screens
with 10% attenuation, and Bottle #7 has 7 screens with 5% attenuation. Cover the bottoms of the bottles so that no light can enter.
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Experiment Procedure
•
14
AP
Biology
EDVO-Kit # 292
Part B: Primary Productivity
Lab #
12
Dissolved Oxygen & Aquatic Primary Productivity
6.
NOTE:
Each group will be assigned
one of the following number
of screens for Bottles 3, 4,
& 5.
Record the starting time here. ________________.
7.
Experiment Procedure
Percent Number of
Light
Screens
100
65
25
10
2
0
1
3
5
8
Put bottles #3-7, labels down, on their sides exposed to a constant source of light. Your teacher will indicate where they should
be placed. Leave these bottles overnight.
You will fix the amount of dissolved oxygen in the Bottle labeled
‘#1 Initial’. This serves as the starting level of dissolved oxygen in
the lake water sample. You will perform step 3 from the Winkler
procedure used in Part A of this laboratory.
•
Open the container labeled “#1 Initial” and carefully pipet
2 ml of manganese sulfate into the container. Make sure the
pipet tip is below the surface of the water in the container.
•
With a fresh pipet, add 2ml of the NaOH/NaI (alkaline-Iodide)
solution into the sample in the container. Make sure the pipet
tip is below the surface of the water in the container.
•
Stopper or seal your container. Firmly hold the stopper or lid of
your container. Carefully invert the bottle to allow for complete mixing of the sample, manganese sulfate, and NaOH/
NaI. A precipitate will form.
•
Let the bottle sit on the laboratory bench overnight. Finish
processing it with the rest of the bottles tomorrow.
Optional Activity
If directed by the instructor, place several drops of the pond water on
a microscope slide and cover with a coverslip. Observe the organisms
found in the natural water sample. Draw what is observed and try to
identify the organisms.
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
15
AP
Part B: Primary Productivity
Biology
Lab #
Day Two
1.
Fix the dissolved oxygen in Bottles #2, 3, 4 , 5, 6 , 7. This is the beginning of the Winkler procedure.
12
Record the time you begin here. __________
•
•
•
2.
Repeat procedure in step 1 above with Bottles #3, 4, 5, 6, 7.
3.
While precipitate is settling in Bottles #2, 3, 4, 5, 6 and 7, review the
Winkler procedure used in Part A. Obtain Bottle #1.
4.
You should clean and set up the burettes as outlined in Part A,
steps 4-7.
5.
Begin processing all of the Bottles, #1 - #7, using the Winkler procedure for the determination of dissolved oxygen from Part A. Part A
Step 3, the fixing of the dissolved oxygen, has already been completed for all of the bottles. Now continue with Part A Step 8. Continue through steps 16. Record DO values in the appropriate table Table 2 or 3.
TABLE 2: Respiration
Individual Data
Class Mean
DO, Initial
DO, Dark Bottle
Respiration Rate
(Initial - Dark)
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EVT 005057AM
Experiment Procedure
Open Bottle #2 and carefully pipet 2ml of manganese sulfate into
the bottle. Make sure pipet tip is below surface of the water in
the container.
With a fresh pipet, add 2ml of the NaOH /NaI (alkaline-iodide)
solution to the sample in the bottle. Make sure pipet tip is below
surface of water in the container.
Stopper or seal the bottle. Firmly hold the stopper or lid and
carefully invert the bottle to allow for complete mixing of the
sample, Manganese sulfate, and NaOH/NaI. A precipitate will
form.
16
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Part B: Primary Productivity
Lab #
12
TABLE 3: Productivity of Screen-Wrappen Samples
Experiment Procedure
# of
Screens
%
Light
0
100
1
65
3
25
5
10
7
5
DO
INDIVIDUAL DATA
Gross
Productivity
6.
Net
Productivity
CLASS MEAN PRODUCTIVITY
DO
Gross
Productivity
Net
Productivity
In reporting the data for productivity, the concentration of dissolved oxygen which has been determined, will be converted
from mg/L to ml/L using the following conversion factor: 1 mg
DO/L = 0.698ml DO/L. Therefore,
[0.698ml D0/L]
X (#mg D0/L) = #ml D0/L
1mg D0/L
7.
Calculate and record the gross and net productivity for the
natural water samples using the following equations. Productivity
is a rate term, therefore, divide by the number of hours the experiment ran to arrive at a value of ml DO/L per hour.
•
Gross Productivity = (Light Bottle (#3, #4, #5, #6, or #7) ml
DO/L - Dark Bottle (#2) ml DO/L)/hours
•
Net Productivity = (Light Bottle (#3, #4, #5, #6. Pr #7) ml DO/L Initial Bottle (#1) ml DO/L) /hours
•
Respiration rate = (Initial Bottle (#1) ml DO/L - Dark Bottle (#2)
ml DO/L) /hours
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
17
AP
Biology
Part B: Primary Productivity
8.
Record the values for the individual experiment conditions here.
•
Respiration rate = __________ml DO/L/hour.
•
Bottle 3
Lab #
12
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
•
Bottle 4
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
Bottle 5
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
•
Bottle 6
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
•
Bottle 7
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
9.
Record the individual and class average values for respiration in
Table 2 and for light of different intensities in the Table 3. The values
should be reported in ml DO/L/hour.
10. Record the average class value for the Respiration rate here.
__________.
11. Using linear graph paper or Graph #2 provided on page 19, plot the
average gross and net productivity on the Y-axis in ml DO/L/hr versus
the percent of Light Intensity on the X-axis.
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EVT 005057AM
Experiment Procedure
•
18
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Analysis of Results - Graph #1
1.
12
Graph both the lab group and class means percent saturation as
function of temperature.
2.
Title the Graph_________________________________________________
3.
Determine the independent variable (horizontal (X) axis). Label
the graph.
4.
Determine the dependent variable (vertical (Y) axis). Label the
graph.
5.
What hypothesis is being tested in this experiment?
Experiment Procedure
Lab #
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
19
AP
Analysis of Results - Graph #2
1.
Graph the average gross and net productivity values for samples as a
function of light intensity (%).
2.
Title the Graph_________________________________________________
3.
Determine the independent variable (horizontal (X) axis). Label the
graph.
4.
Determine the dependent variable (vertical (Y) axis). Label the
graph.
5.
What hypothesis is being tested in this experiment?
Biology
Lab #
12
Experiment Procedure
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EVT 005057AM
20
AP
Biology
Lab #
Experiment Procedure
12
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Laboratory Extensions
I.
EFFECT OF ADDITION OF NITROGEN AND PHOSPHORUS
ON PRODUCTIVITY
The availability of inorganic nutrients such as nitrogen and phosphorous may also affect primary productivity in aquatic environments.
Nutrients are removed during primary productivity. When a nutrient is
no longer present in sufficient quantity, a slowing of productivity will
result. The addition of specific nutrients may result in an increase in
productivity provided all the other necessary nutrients are present in
adequate amounts. As in Part B., samples of natural water will be
used to determine gross productivity, net productivity, and respiration
rates. This experiment will determine the effect of the nutrients nitrogen and phosphorus on primary productivity. Review the Background
section of this laboratory for a complete discussion of Primary Productivity.
Day One
1.
2.
Obtain 5 clean “BOD” (Biological Oxygen Demand) containers.
Any container which holds approximately 250-300ml and which
can be closed with an airtight seal is appropriate.
•
Completely fill each bottle with water sample from the lake or
pond. Allow the sample to overflow the container so that it
will be completely filled.
•
Stopper or close the container. Turn the container upside
down and use a paper towel to remove any water which is
around the outside of the stopper or lid.
Label 5 bottles as follows:
#1 Initial
#2 Dark
#3 Light + Nitrogen Addition
#4 Light + Phosphorous Addition
#5 Light Alone
3.
Bottle #1 is the initial starting bottle and serves as a baseline.
4.
Wrap Bottle #2 in aluminum foil, for it serves as the dark (no light)
control.
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
21
AP
Laboratory Extensions
Biology
Lab #
Bottle #3 is supplemented with a source of nitrogen and exposed to
light. With a pipet, add 1.0ml of nitrogen enrichment solution to Bottle
#3. The instructor may have set up a pipeting station for this step.
6.
Bottle #4 is supplemented with a source of phosphorous and exposed
to light. With a pipet, add 1.0ml of phosphorous enrichment solution
to bottle 4. The instructor may have set up a pipeting station for this
step.
7.
Bottle #5 is a control which receives no nutrient supplementation, but
is exposed to light.
8.
Put Bottle #2 in a dark place and Bottles #3, #4, and #5 so that they
are exposed to a constant source of light. Your teacher will indicate
where they should be placed. Leave these bottles overnight.
Record the starting time here. ________________
9.
You will fix the amount of dissolved oxygen in the bottle labeled
‘#1 Initial’. This serves as the starting level of dissolved oxygen in the
lake water sample. You will perform step 3 from the Winkler procedure used in Part A of this laboratory.
•
Open the container labeled “#1 Initial” and carefully pipet 2ml of
manganese sulfate into the container. Make sure the pipet tip is
below the surface of the water in the container.
•
With a fresh pipet, add 2ml of the NaOH/NaI (alkaline-Iodide)
solution into the sample in the container. Make sure the pipet tip
is below the surface of the water in the container.
•
Stopper or seal your container. Firmly hold the stopper or lid of
your container. Carefully invert the bottle to allow for complete
mixing of the sample, manganese sulfate, and NaOH/NaI. A
precipitate will form.
•
Let the bottle sit on the laboratory bench overnight. Finish
processing it with the rest of the bottles tomorrow.
Optional Activity
If directed by the instructor, place several drops of the pond water on a
microscope slide and cover with a coverslip. Observe the organisms
found in the natural water sample. Draw what is observed and try to
identify the organisms.
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EVT 005057AM
12
Experiment Procedure
5.
22
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
AP
Biology
Lab #
12
Laboratory Extensions
Day Two
1.
Fix the dissolved oxygen in Bottles #2, #3, #4 and# 5. This is the
beginning of the Winkler procedure.
Record the time you begin here. __________
•
•
Experiment Procedure
•
Open Bottle #2 and carefully pipet 2ml of manganese sulfate
into the container. Make sure pipet tip is below surface of the
water in the container.
With a fresh pipet, add 2ml of the NaOH /NaI (alkaline-iodide)
solution into the sample in the bottle. Make sure pipet tip is
below surface of water in the container.
Stopper or seal the bottle. Firmly hold the stopper or lid and
carefully invert the bottle to allow for complete mixing of the
sample, Manganese sulfate, and NaOH/NaI. A precipitate will
form.
2.
Repeat procedure in step 1 above with Bottles #3, #4, and #5.
3.
While precipitate is settling in Bottles #2, #3, #4, and #5, review the
Winkler procedure used in Part A. Obtain Bottle #1.
4.
You should clean and set up the burettes as outlined in Part A,
steps 4-7.
5.
Begin processing all of the Bottles #1, #2, #3, #4, and #5 using the
Winkler procedure for the determination of dissolved oxygen from
Part A. Part A Step 3 , the fixing of the dissolved oxygen, has
already been completed for all of the bottles. Now continue with
Part A Step 8.
6.
In reporting the data for productivity, the concentration of dissolved oxygen which has been determined, will be converted
from mg/L to ml/L using the following conversion factor: 1 mg
DO/L = 0.698ml DO/L. Therefore,
[0.698ml D0/L]
1mg D0/L
7.
X (#mg D0/L) = #ml D0/L
Calculate and record the gross and net productivity for the
natural water samples using the following equations. Productivity
is a rate term, therefore, divide by the number of hours the
experiment ran to arrive at a value of ml DO/L per hour.
•
•
•
Gross Productivity = (Light Bottle (#3, #4, or #5) ml DO/L Dark Bottle (#2) ml DO/L)/hours
Net Productivity = (Light Bottle (#3, #4, or # 5) ml DO/L - Initial
Bottle (#1) ml DO/L) /hours
Respiration rate = (Initial Bottle (#1) ml DO/L - Dark Bottle (#2)
ml DO/L) /hours
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
23
AP
Biology
Laboratory Extensions
8.
Record the values for the individual experiment conditions here.
•
Respiration rate = __________ml DO/L/hour.
•
Nitrogen enriched Bottle 3
Lab #
12
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
•
Phosphorous enriched bottle 4
Gross Productivity rate = ___________ ml DO/L/hour
•
Un-enriched Bottle 5
Gross Productivity rate = ___________ ml DO/L/hour
Net Productivity rate = ____________ ml DO/L/hour
9.
Record the class average values for Respiration Rate in Table 4 and
for nitrogen and phosphorous enriched samples in Table 5. The values
should be reported in ml DO/L/hour.
10. Record the average class value for the Respiration rate here.
__________.
11. Using linear graph paper or Graph #3 on page 25, plot the average
gross and net productivity on the Y-axis in ml DO/L/hr versus nitrogen
enriched, phosphorous enriched, and un-enriched samples on the Xaxis.
TABLE 4: Respiration
Individual Data
Class Mean
DO, Initial
DO, Dark Bottle
Respiration Rate
(Initial - Dark)
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EVT 005057AM
Experiment Procedure
Net Productivity rate = ____________ ml DO/L/hour
24
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
AP
Biology
Laboratory Extensions
Lab #
Experiment Procedure
12
TABLE 5: Productivity of Pond Samples - Nitrogen or Phosphorous Added
Sample
Treatment
%
Light
Nitrogen
Added
100
Phosphorous
Added
100
Unenriched
100
Little Winter Pond
% Light
Depth in
Intensity
Meters
100
0
65
25
0.3
1.0
10
2
3.0
5.0
INDIVIDUAL DATA
DO
Gross
Productivity
Intensity
Meters
100
0
65
25
3.5
9
10
2
15
25
DO
Gross
Productivity
Net
Productivity
From the results in Part B of this laboratory, compare the productivity
of two separate ponds located in Massachusetts. This exercise is a
simulation. In Little Winter Pond, water is murky and filled with algae
and weeds. Light does not penetrate far beneath the surface. In Big
Winter Pond, a clear pond high in the mountains, light is not attenuated much until you get to a very deep spot near the middle of the
lake. The Little and Big Winter Pond have the values as shown in the
Tables at left. The values for these two hypothetical ponds will be
used for plotting the data.
Big Winter Pond
Depth in
Net
Productivity
II. PRODUCTIVITY SIMULATION FROM 2 PONDS
1.
% Light
CLASS MEAN PRODUCTIVITY
To simulate the productivity in the two ponds, convert your gross
productivity data for the un-enriched sample (5) in ml DO/L/hour
to ml DO/L/day. Simply multiply by 24 to get the value per day.
# ml DO/L
hour
2.
X
24 hour
day
=
# ml DO/L
day
Next, convert the gross productivity values in ml DO/L/day to
carbon productivity in mg C/m3/day. For each ml of oxygen
produced, 0.536 mg of carbon has been consumed. Therefore,
simply multiply the ml DO/L/day values by 0.536 to get the carbon
productivity in mg C/m3/day. Also convert the respiration rate to
carbon values, again in mg C/m3/day.
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
25
AP
Analysis of Results - Graph #3
1.
Graph the average gross and net productivity values for nitrogen
enriched, phosphorous enriched and un-enriched samples.
2.
Title the Graph_________________________________________________
3.
Determine the independent variable (horizontal (X) axis). Label the
graph.
4.
Determine the dependent variable (vertical (Y) axis). Label the
graph.
5.
What hypothesis is being tested in this experiment?
Biology
Lab #
12
Experiment Procedure
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EVT 005057AM
26
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Optional Analysis of Results - Graph #4
1.
12
Graph the converted carbon assimilation values versus depth
related to light intensity for Little and Big Winter ponds.
2.
Title the Graph_________________________________________________
3.
Determine the independent variable (horizontal (X) axis). Label
the graph.
4.
Determine the dependent variable (vertical (Y) axis). Label the
graph.
5.
What hypothesis is being tested in this experiment?
Experiment Procedure
Lab #
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
27
AP
Biology
Study Questions
How does temperature affect the solubility of oxygen in water?
2.
How does salinity affect the solubility of oxygen in water?
3.
Would you expect boiled water to contain a large amount of dissolved oxygen ?
4.
How many moles of sodium thiosulfate are required to completely
titrate 6 moles of oxygen.
5.
Why is the starch usually added after the titration has begun?
6.
From Graph #2 (page 19), were any of the samples from Part B limited
by the available light?
7.
Were any samples limited by the available nutrients?
8.
Explain why the DO concentration of water samples taken from a
lake in early morning would be lower than the DO concentration of
water samples taken in late afternoon.
9.
Explain why the DO concentration of water samples taken from a
swiftly flowing stream would be higher than the DO concentration of
water samples taken from a lake.
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EVT 005057AM
Lab #
12
Experiment Procedure
1.
28
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Notes:
Lab #
Experiment Procedure
12
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EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
AP
Notes to the Instructor
Biology
OVERVIEW OF LABORATORY INVESTIGATIONS
The "hands-on" laboratory experience is a very important component of
the science courses. Laboratory experiment activities allow students to
identify assumptions, use critical and logical thinking, and consider
alternative explanations, as well as help apply themes and concepts to
biological processes.
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Class size, length of laboratory sessions, and availability of equipment
are factors which must be considered in the planning and the implementation of this experiment with your
students. These guidelines can be adapted
to fit your specific set of circumstances.
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EVT 005057AM
Lab #
12
Instructor's Guide
EDVOTEK experiments have been designed to provide students the
opportunity to learn very important concepts and techniques used by
scientists in laboratories conducting biotechnology research. Some of
the experimental procedures may have been modified or adapted to
minimize equipment requirements and to emphasize safety in the
classroom, but do not compromise the educational experience for
the student. The experiments have been tested repeatedly to
maximize a successful transition from the laboratory to the classroom
setting. Furthermore, the experiments allow teachers and students
the flexibility to further modify and adapt procedures for laboratory
extensions or alternative inquiry-based investigations.
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29
30
AP
Biology
Lab #
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Pre-Lab Preparations
•
Part A requires 2 liters each of fresh tap water at 5°, 20°, and 30°C.
The water should be held covered at these temperatures the day
before the laboratory.
•
A natural source of water such as lake, pond, seawater, or an
algal culture of Chlorella will work for Part B. Depending on the
size of the bottles or flasks to be used, collect 15 liters of natural
source water.
•
The solutions should be prepared 1 or 2 days before the laboratory. The sodium thiosulfate should be prepared the day before
and stored in the refrigerator. Working stocks are made on the
day of the laboratory.
12
Instructor's Guide
Part A. The Winkler Method for Dissolved Oxygen
1.
Preparation of the sodium thiosulfate solution:
•
Boil 4 liters of distilled water for 10-15 minutes (to be completed before lab session).
•
On the day of the first laboratory session, add all of the
sodium thiosulfate (component E) to boiled and cooled
distilled water to make 2 liters final volume. Boiling depletes
the oxygen in the water so that it will not interfere with the
analysis.
•
Store carefully sealed in the refrigerator.
2.
The thiosulfate solution will not be standardized since fine “analytical” results are not needed. When the thiosulfate is properly
measured on a balance, titration is not necessary for classroom
results.
3.
Just prior to the titration step in Part A, Student Experimental
Procedures:
4.
•
Prepare the sodium thiosulfate working solution for the class.
•
Add 250ml of the working stock solution to 750ml of boiled
and cooled distilled water.
On the day of the lab, set up measuring/pipeting stations for:
•
•
•
•
•
•
Tap water (250-300ml)
Manganese Sulfate (2ml)
NaOH/NaI (2ml)
Sodium thiosulfate (50ml)
Concentrated sulfuric acid (2ml)
Starch solution (1ml)
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Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/
laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose
without the written consent of EDVOTEK, Inc. Copyright © 1996,1997, 1998, 1999, 2000, 2001, 2005,2007
EDVOTEK, Inc., all rights reserved.
EVT 005057AM
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
31
AP
Biology
Pre-Lab Preparations
Lab #
5.
6.
Prepare the manganese sulfate solution:
•
Add all of the manganese sulfate (component C) to a total
volume of 250ml of distilled water.
•
Filter with #1 filter paper and store covered at room temperature.
Wearing gloves and goggles, prepare the NaI/NaOH solution:
Add all of the NaOH pellets (component B) to a large beaker.
•
Add distilled water to a volume of 150 - 175ml. Caution: The
solution will get hot.
•
Add all of the NaI (component A) and adjust to a final volume of
250ml with distilled water. Mix.
•
Store covered at room temperature.
2.
3.
4.
On the day of the lab, prepare a working nitrogen enrichment solution
for the class:
•
Add 10ml of the stock solution (component F) to 500ml of distilled
water.
•
Store covered in the refrigerator. Students will use 1ml of the
working solution to provide nitrogen enrichment for their sample.
Prepare the phosphorous enrichment stock solution:
•
Add all the contents of component G to 100ml distilled water. Mix.
•
Store covered in the refrigerator.
On the day of the lab, prepare a working phosphorous enrichment
solution for the class:
•
Add 10ml of stock solution to 500ml of distilled water.
•
Students will use 1ml of the working solution to provide phosphorous enrichment for their sample.
Part B will require 50 BOD type bottles (250-300ml). You may substitute
1-800-EDVOTEK • www.edvotek.com
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/
laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose
without the written consent of EDVOTEK, Inc. Copyright © 1996,1997, 1998, 1999, 2000, 2001, 2005,2007
EDVOTEK, Inc., all rights reserved.
EVT 005057AM
Instructor's Guide
•
Part B. Primary Productivity
1.
12
32
AP
Biology
EDVO-Kit # 292
Dissolved Oxygen & Aquatic Primary Productivity
Pre-Lab Preparations
Lab #
12
250ml Erlenmeyer flasks. Small juice bottles with caps can also be
used successfully.
5.
Based on Table 1, assign each group a different light percentage
(corresponding number of screens). The screens will be used to
cover Bottles 3, 4, and 5.
6.
On the day of the lab, set up measuring/pipeting stations for:
•
•
•
•
•
Instructor's Guide
7.
Pond water (250-300ml)
Nitrogen enrichment solution (1ml)
Phosphorous enrichment solution (1ml)
Manganese sulfate (2ml)
NaOH/NaI (2ml)
Set up microscopes if the optional activity in Student Experimental
Procedures B. #11 is to be done by students.
1-800-EDVOTEK • www.edvotek.com
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/
laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose
without the written consent of EDVOTEK, Inc. Copyright © 1996,1997, 1998, 1999, 2000, 2001, 2005,2007
EDVOTEK, Inc., all rights reserved.
EVT 005057AM
Please refer to the kit
insert for the Answers to
Study Questions
Section V - Reactivity Data
Material Safety Data Sheet
IDENTITY (As Used on Label and List)
Emergency Telephone Number
Manufacturer's Name
EDVOTEK, Inc.
Incompatibility
(301) 251-5990
(301) 251-5990
Date Prepared
14676 Rothgeb Drive
Rockville, MD 20850
09-18-2002
ACGIH TLV
Sulfur oxides
Conditions to Avoid
May Occur
X
Will Not Occur
Section VI - Health Hazard Data
Inhalation?
Ingestion?
Skin?
Yes
Yes
Yes
Causes eye and skin irritation. Material is irritating to mucous
membranes and upper respiratory tract.
Health Hazards (Acute and Chronic)
IARC Monographs?
NTP?
OSHA Regulation?
No data
Signs and Symptoms of Exposure A stolid mask-like appearance of face, sleepiness d weakness
skin and eye irritation, irritating to mucous membranes and upper respiratory
Section II - Hazardous Ingredients/Identify Information
OSHA PEL
Hazardous
Polymerization
Carcinogenicity:
Signature of Preparer (optional)
Hazardous Components [Specific
Chemical Identity; Common Name(s)]
Strong acids
Hazardous Decomposition or Byproducts
Route(s) of Entry:
Telephone Number for information
Address (Number, Street, City, State, Zip Code)
X
Stable
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Manganese Sulfate Monohydride
Section I
Conditions to Avoid
Unstable
Stability
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200 Standard must be consulted for
specific requirements.
®
Medical Conditions Generally Aggravated by Exposure
Other Limits
Recommended
% (Optional)
Emergency First Aid Procedures
CAS # 10034-96-5
Flush eyes or skin with copious amounts of water for atleast 15 minutes while
removing contaminated clothing and shoes. Remove to fresh air.
Section VII - Precautions for Safe Handling and Use
Section III - Physical/Chemical Characteristics
Boiling Point
No data
Vapor Pressure (mm Hg.)
No data
Vapor Density (AIR = 1)
No data
Steps to be Taken in case Material is Released for Spilled
Specific Gravity (H 0 = 1)
2
2.950
Melting Point
No data
Evaporation Rate
(Butyl Acetate = 1)
No data
Sweep up, place in a bag and hold for waste disposal. Avoid raising dust.
Ventilate area and wash soil site after material pick up is complete.
Waste Disposal Method
The material should be dissolved in 1) water 2) acid solution 3) Oxidized to a water-soluble state.
Solubility in Water
Precautions to be Taken in Handling and Storing
Keep tightly closed , wash thoroughly after handling
Appearance and Odor
Other Precautions
White to pale-pink powder
None
Section IV - Physical/Chemical Characteristics
Flash Point (Method Used)
LEL
Flammable Limits
No data
UEL
Extinguishing Media
Noncombustible-use extinquishing media appropriate to surrounding fire condition
Special Fire Fighting Procedures
Section VIII - Control Measures
Respiratory Protection (Specify Type)
X
Eye Protection
Chemical resistant gloves
Other Protective Clothing or Equipment
Emits toxic fumes under fire conditions
Other
Mechanical (General)
Protective Gloves
Unusual Fire and Explosion Hazards
Special
Yes
Local Exhaust
Ventilation
Wear SCBA and protective clothing to prevent contact with skin and eyes.
Safety goggles
SCBA, safety gloves and goggles
Work/Hygienic Practices
Material Safety Data Sheet
Section V - Reactivity Data
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200. Standard must be consulted for
specific requirements.
Stability
Unstable
IDENTITY (As Used on Label and List)
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Sodium Iodide
Section I
14676 Rothgeb Drive
Emergency Telephone Number
301-251-5990
Telephone Number for information
301-251-5990
Rockville, MD 20850
Date Prepared
09-19-2002
EDVOTEK
Address (Number, Street, City, State, and ZIP Code)
Conditions to Avoid
Stable
Incompatiblity (Materials to avoid)
Manufacturer's Name
NIOSH/MSHA approved respirator
X
Incompatibles
Alkali metals, bromine triflouride, strong oxidizers, perchloric acid, perchloryl
Hazardous Decomposition or Byproducts
thermal decomposition products may include toxic and corrosive fumes of iodides
May Occur
Hazardous
Polymerization
Will Not Occur
X
Section VI - Health Hazard Data
Route(s) of Entry:
Inhalation?
Skin?
Yes
Yes
Ingestion?
Yes
Health Hazards (Acute and Chronic)
May cause irritation. Chronic ingestion may result in hypothyroidism.
Signature of Preparer (optional)
Carcinogenicity:
NTP?
IARC Monographs?
None
OSHA Regulation?
No Data
Section II - Hazardous Ingredients/Identity Information
Hazardous Components
(Specific Chemical Identity; Common Name(s))
OSHA PEL
CAS # 7681-62-5
No Data
ACGIH TLV
Other Limits
Recommended
Signs and Symptoms of Exposure
% (Optional)
Inhalation: mucous membrane irritation Eye/skin contact: flush w/ water
Medical Conditions Generally Aggravated by Exposure
No data
Contains no hazardous components
Emergency and First Aid Procedures
Treat symptomatically and supportively. Eye/skin contact: flush w/ water
Inhalation: remove to fresh air.
Section III - Physical/Chemical Characteristics
Boiling Point
1304ϒC
1.91
Vapor
Pressure (mm Hg.)
Specific Gravity (H20 = 1)
Section VII - Precautions for Safe Handling and Use
at 25ϒC
Melting Point
1
Vapor Density (AIR =1)
N.D. = No Data
N.D.
3.667
Steps to Be Taken in case Material Is Released or Spilled
Sweep up, place in a bag and hold for waste disposal. Avoid raising dust.
661ϒC
Evaporation Rate
(Butyl Acetate =1)
N.D.
Ventilate area and wash spill site after pickup.
Waste Disposal Method
Follow federal, state and local regulations.
Solubility in Water
184% at 25ϒC
Precautions to be Taken in Handling and Storing
Appearance and Odor
odorless, white powder
Section IV - Fire and Explosion Hazard Data
Flash Point (Method Used)
Extinguishing Media
Flammable Limits
Other Precautions
LEL
Store away from incompatibles
Avoid contact
UEL
Section VIII - Control Measures
Use suitable agent for type of surrounding fire
Special Fire Fighting Procedures
Move container from fire area if possible. Avoid breathing vapors or dust.
Unusual Fire and Explosion Hazards
None
Respiratory Protection (Specify Type)
NIOSH/MSHA approved respirator
Special
Yes
No
Mechanical (General) No
Other
None
Protective Gloves
Eye Protection Safety goggles
PVC gloves
Other Protective Clothing or Equipment
Impervious clothing and equipment to prevent contact
Ventilation
Work/Hygienic Practices
Local Exhaust
Avoid contact
Section V - Reactivity Data
Material Safety Data Sheet
IDENTITY (As Used on Label and List)
Manufacturer's Name
EDVOTEK, Inc.
(301) 251-5990
(301) 251-5990
Date Prepared
14676 Rothgeb Drive
Rockville, MD 20850
09-19-2002
Sodium Hydroxide
CAS # 1310-73-2
2mg/m3
ACGIH TLV
Ingestion?
Skin?
Yes
Yes
Yes
None identified
IARC Monographs?
NTP?
OSHA Regulation?
None identified
Call physician. Ingestion: Do not induce vomiting. Give water followed by vinegar, juice or egg white
Inhalation: Move to fresh air. Skin/eye contact: flush with water
Section VII - Precautions for Safe Handling and Use
Steps to be Taken in case Material is Released for Spilled
1390°C
Specific Gravity (H 0 = 1)
2
2.13
Vapor Pressure (mm Hg.)
20°C
Melting Point
318°C
NO data
Evaporation Rate
(Butyl Acetate = 1)
NO data
Dispose of properly
Wear SCBA and protective clothing. Carefully place material into clean, dry container and cover.
Waste Disposal Method
Follow all federal, state, and local laws.
Precautions to be Taken in Handling and Storing
Keep container tightly closed. Store in corrosion-proof area. Store in a dry area.
Isolate from incompatible materials.
10% appreciable
Other Precautions
Appearance and Odor
White pellets, odorless
None
Section IV - Physical/Chemical Characteristics
Flash Point (Method Used)
Flammable Limits
LEL
UEL
NA
Extinguishing Media
Inhalation?
Health Hazards (Acute and Chronic)
Emergency First Aid Procedures
No data
Boiling Point
Solubility in Water
Section VI - Health Hazard Data
Medical Conditions Generally Aggravated by Exposure
% (Optional)
Section III - Physical/Chemical Characteristics
Vapor Density (AIR = 1)
X
No data
No data
No data
Signs and Symptoms of Exposure Ingestion: Severe burns to mouth, throat, and stomach, nausea & vomiting
Inhalation: irritation Skin/eye contact: severe irritation or burns
Other Limits
Recommended
2mg/m3
Conditions to Avoid
May Occur
Will Not Occur
None
Section II - Hazardous Ingredients/Identify Information
OSHA PEL
Hazardous
Polymerization
Carcinogenicity:
Signature of Preparer (optional)
Hazardous Components [Specific
Chemical Identity; Common Name(s)]
Water, strong acids, metals, combustible materials, organic materials
Zinc, aluminous, peroxide, halogenated hydroca
Hazardous Decomposition or Byproducts
None identified
Route(s) of Entry:
Telephone Number for information
Address (Number, Street, City, State, Zip Code)
Moisture
Incompatibility
Section I
Emergency Telephone Number
X
Stable
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Sodium Hydroxide
Conditions to Avoid
Unstable
Stability
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200 Standard must be consulted for
specific requirements.
®
NA
NA
Section VIII - Control Measures
Respiratory Protection (Specify Type)
NIOSH/MSHA approved respirator
Yes
Use extinquishing media appropriate for surrounding fire
Yes
Mechanical (General)
Special Fire Fighting Procedures
Protective Gloves
Wear protective equipment and self-contained breathing apparatus. Floof material with water
Contact with moisture or water generate sufficient heat to ignite other materials.
React with metals to produce hydrogen gas which can form explosive mixture with air.
Material Safety Data Sheet
Work/Hygienic Practices
Other
Eye Protection
Neoprene gloves
Other Protective Clothing or Equipment
Unusual Fire and Explosion Hazards
Special
Local Exhaust
Ventilation
No
None
Safety goggles
Uniform, apron
Avoid contact
Section V - Reactivity Data
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200. Standard must be consulted for
specific requirements.
Stability
Unstable
Conditions to Avoid
Stable
X
incompatibles
Incompatiblity (Materials to avoid)
Strong acids, strong oxidizing agents, lead, silver and mercury salts, and iodines
IDENTITY (As Used on Label and List)
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Sodium Thiosulfate
Section I
Manufacturer's Name
14676 Rothgeb Drive
Emergency Telephone Number
301-251-5990
Telephone Number for information
301-251-5990
Rockville, MD 20850
Date Prepared
09-19-2002
EDVOTEK
Address (Number, Street, City, State, and ZIP Code)
Hazardous Decomposition or Byproducts
Sulfur oxides
May Occur
Hazardous
Polymerization
Will Not Occur
X
Incompatibles
Section VI - Health Hazard Data
Route(s) of Entry:
Inhalation?
Signature of Preparer (optional)
Skin?
Yes
Health Hazards (Acute and Chronic)
Carcinogenicity:
Yes
Ingestion?
Yes
Eye/skin irritation
NTP?
IARC Monographs?
OSHA Regulation?
No data
Section II - Hazardous Ingredients/Identity Information
Hazardous Components
(Specific Chemical Identity; Common Name(s))
OSHA PEL
ACGIH TLV
Other Limits
Recommended
% (Optional)
CAS # 10102-17-7
Signs and Symptoms of Exposure
Irritation
Medical Conditions Generally Aggravated by Exposure
No data
Does not contain any hazardous components
Emergency and First Aid Procedures
Sodium thiosulfate pentahydrate
Skin/eye contact flush with copious amounts of H20 for at least 15 minutes . Remove to fresh air &
remove contaminated clothing
Section III - Physical/Chemical Characteristics
Boiling Point
N.D.
Vapor Pressure (mm Hg.)
N.D.
Vapor Density (AIR =1)
N.D.
Section VII - Precautions for Safe Handling and Use
Specific Gravity (H20 = 1)
1.729
Melting Point
Evaporation Rate
(Butyl Acetate =1)
N.D.
Water soluble
Waste Disposal Method
Cautiously add to a large stirred excess of water, adjust pH to neutral. Flush the aqueous solution down
Precautions to be Taken in Handling and Storing
White crystals, no odor
Section IV - Fire and Explosion Hazard Data
Flammable Limits
UEL
Wear SCBA and protective clothing to prevent contact with skin and eyes
Emits toxic fumes under fire conditions.
Avoid incompatibles
Other Precautions
LEL
Use media for surrounding fire
Special Fire Fighting Procedures
Unusual Fire and Explosion Hazards
Ventilate area and wash spill site after pickup.
drain with plenty of water.
Appearance and Odor
Extinguishing Media
Sweep up, place in a bag and hold for waste disposal. Avoid raising dust.
N.D.
Solubility in Water
Flash Point (Method Used)
Steps to Be Taken in case Material Is Released or Spilled
Do not breathe dust. Avoid contact.
Section VIII - Control Measures
Respiratory Protection (Specify Type)
NIOSH/MSHA approved respirator
No
No
Special
Mechanical (General) Yes
Other
None
Protective Gloves
Eye Protection Safety goggles
Chemical resistant
Other Protective Clothing or Equipment
None
Ventilation
Work/Hygienic Practices
Local Exhaust
Wash thoroughly after handling
Material Safety Data Sheet
Section V - Reactivity Data
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200. Standard must be consulted for
specific requirements.
Stability
Unstable
Conditions to Avoid
Stable
X
Incompatiblity (Materials to avoid)
IDENTITY (As Used on Label and List)
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Nitrogen Enrichment Solution
Section I
Manufacturer's Name
Emergency T
Telephone Number
301-251-5990
Telephone Number for information
T
301-251-5990
EDVOTEK
Address (Number, Street, City, State, and ZIP Code)
Rockville, MD 20850
Incompatibles
Strongg reducingg agents,
g
, metals,, stron acids,, strongg bases
Hazardous Decomposition or Byproducts
May Occur
Hazardous
Polymerization
X
Incompatibles
Section VI - Health Hazard Data
Route(s) of Entry:
Skin?
es
Ingestion?
g
Ys
Ye
Yes
Ye
Mayy cause mucous membranes
b
and upper
pp respiratory
p
y tract irritation. Eye/skin
y
irritation
0
09-15-02
Signature of Preparer (optional)
Carcinogenicity:
NTP?
IARC Monographs?
OSHA Regulation?
No Data
Hazardous Components
(Specific Chemical Identity; Common Name(s))
OSHA PEL
ACGIH TLV
L
LV
Other Limits
Recommended
% (Optional)
Signs and Symptoms of Exposure
Irritation
CAS # 7757-79-1
CAS# 12125-02-9
No Data
Contains no hazardous components
p
Emergency and First Aid Procedures
N.D.= Not Determined
Boiling Point
N.D.
Vapor Pressure (mm Hg.)
V
N.D.
N.D.
Specific Gravity (H20 = 1)
N.D.
Melting Point
Avoid raisingg dust.
A
N.D.
Evaporation Rate
(Butyl
(B
t lA
Acetate
t t =1)
1
N.D.
W
Solubility in Water
Observe all federal, state, and local regulations
soluble
Appearance and Odor
Clear liquid, characteristic odor
Section IV - Fire and Explosion Hazard Data
Flash Point (Method Used)
Precautions to be T
Taken in Handling and Storing
Flammable Limits
LEL
Other Precautions
Extinguishing Media
Keep away from incompatibles
UEL
Avoid contact
A
Water spray
W
Special Fire Fighting Procedures
Wear
W
ear SCBA
A and pprotective clothing
g to pprevent contact with skin and eyes
y
Respiratory Protection (Specify Type)
T
Ventilation
No
Local Exhaust
Special
es
Unusual Fire and Explosion Hazards
Emits toxic fumes. Contact with other material mayy cause fire.
Protective Gloves
Chemical resistant
Work/Hygienic Practices
No
Other
None
Eye Protection Safety goggles
Avoid contact and incompatibles
A
Section V - Reactivity Data
Material Safety Data Sheet
IDENTITY (As Used on Label and List)
Emergency Telephone Number
Manufacturer's Name
EDVOTEK, Inc.
Incompatibility
(301) 251-5990
(301) 251-5990
Date Prepared
14676 Rothgeb Drive
Rockville, MD 20850
Incompatibles
Alkaloids, antipyrine, choral hydrate, lead acetate, pyrogallol, resorcinol
Hazardous Decomposition or Byproducts
Hazardous
Polymerization
phosphorous oxides, sodium oxide
Conditions to Avoid
May Occur
X
Will Not Occur
None
Section VI - Health Hazard Data
Route(s) of Entry:
Telephone Number for information
Address (Number, Street, City, State, Zip Code)
X
Stable
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Phosphorous Enrichment Reagents
Section I
Conditions to Avoid
Unstable
Stability
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200 Standard must be consulted for
specific requirements.
®
Inhalation?
Ingestion?
Skin?
Yes
Yes
Yes
Health Hazards (Acute and Chronic)
None
09-19-2002
Carcinogenicity:
IARC Monographs?
NTP?
Signature of Preparer (optional)
OSHA Regulation?
No data
Signs and Symptoms of Exposure
Section II - Hazardous Ingredients/Identify Information
Hazardous Components [Specific
Chemical Identity; Common Name(s)]
OSHA PEL
ACGIH TLV
Other Limits
Recommended
Medical Conditions Generally Aggravated by Exposure
% (Optional)
Inhalation: remove to fresh air Skin/eye contact: flush with water
Ingestion: treat symptomatically and supportively. Seek medical attention
Section VII - Precautions for Safe Handling and Use
Section III - Physical/Chemical Characteristics
Boiling Point
Steps to be Taken in case Material is Released for Spilled
Specific Gravity (H 0 = 1)
2
decomposes
Vapor Pressure (mm Hg.)
No data
Vapor Density (AIR = 1)
No data
Solubility in Water
No data
Melting Point
Evaporation Rate
(Butyl Acetate = 1)
No data
Keep container closed and store away from incompatibles
Avoid contact
Flammable Limits
LEL
UEL
Extinguishing Media
Water spray, CO2, Dry chemical powder or appropriate foam
Special Fire Fighting Procedures
Wear SCBA and protective clothing to prevent contact with skin and eyes
Move container from fire area if possible. Do not scatter spilled material with high pressure water
Unusual Fire and Explosion Hazards
Avoid breathing fumes
Observe all federal, state, and local regulations
Other Precautions
Odorless, white powder
No data
Waste Disposal Method
Precautions to be Taken in Handling and Storing
Section IV - Physical/Chemical Characteristics
Flash Point (Method Used)
Sweep up, place in a bag and hold for waste disposal. Avoid raising dust.
Ventilate area and wash spill site after pickup.
No data
Soluble
Appearance and Odor
No Data
Emergency First Aid Procedures
CAS # 7558-79-4
Contains no hazardous components
Section VIII - Control Measures
Respiratory Protection (Specify Type)
Ventilation
Protective Gloves
NIOSH/MSHA approved respirator
Local Exhaust
No
Mechanical (General)
Yes
Work/Hygienic Practices
Other
Eye Protection
YEs
Other Protective Clothing or Equipment
Special
Wear equipment to prevent contact
Avoid contact
No
None
Safety goggles