Harnessing Algae as a Biofuel Lab Report final copy
1. Harnessing Algae as a Biofuel
CE 420 - Water Quality Engineering Laboratory
Fall Semester 2015, Thursday Section
Kim Colavito, Mikhail Thomas, Alison Koerkenmeier
Due: December 18, 2015
2. Executive Summary
Algae is a green, marine-based, photosynthesizing organism that grows/lives in water and creates
energy from sunlight ("All About Algae Supplements | Precision Nutrition."). It is categorized in
a group of its own because it does not have the characteristics as land-plants do: leaves and roots.
There are many different forms of algae that have their own distinct properties.
The valuable aspect of algae is its ability to be used for an alternative fuel source by extracting
the lipids and crude oils produced. The objective of this study was to develop an efficient method
of growing algae in order to make production of biofuel from algae enticing for enterprising for
commercial use. The study attempted to mimic the natural conditions under which algae blooms
flourish as closely as possible.
In order to mimic the natural conditions of algae, only nutrients that occurred naturally that
affected algae growth were analyzed: nitrate and phosphate. While they are both found naturally
in the environment, nitrogen is far more abundant, constituting 78% of the air on the planet.
Thus, the turnover of the nitrogen cycle is far quicker than that of phosphorus which can be
found mostly in rocks and minerals. Thus phosphorus is often the limiting factor in the growth of
aquatic plants and other organisms.
There was a flaw in this method of thinking, however, because this study also analyzed different
types of algae feed and the initial algae was grown in a nutrient rich incubation tank with algae
feeds of BG-11 Medium and F/2 Part A and B Solution. These algae feeds do not occur naturally
in algae’s environment; however the feed was used to spark algae growth as the study occurred
in a limited time period.
Despite this, the levels optimum levels of nitrate and phosphate were still analyzed in order to
determine the concentrations of both that produce optimal algae growth. To determine these
concentrations, algae was grown in varying concentrations of both nitrogen and phosphorus, and
the effects on each on algae growth determined by weight of algae. The best method to measure
this was by using a suspending solids test was used to determine the weight of algae in each test
condition by comparing initial and successive weights of the algae over time.
The nutrient concentrations of each sample were also determined through the use of an Ion
Chromatograph, and the correlation between the weight of solids obtained and the nutrient level
absorbed by the algae was determined and displayed.
The procedure and results of both the suspended solids tests and the Ion Chromatograph tests are
contained within this report, as well as a final recommendation of what optimal food and nutrient
concentrations are needed to achieve the optimal growth of algae.
3. Table of Contents
Executive Summary .........................................................................................................................................................................2
Table of Contents .............................................................................................................................................................................3
1.0 Introduction...............................................................................................................................................................................4
1.1 Literature Review.................................................................................................................................................................4
1.2 Experimental Objective.......................................................................................................................................................5
2.0 Materials and Methods............................................................................................................................................................6
2.1 Experimental Design............................................................................................................................................................6
2.2 Experiment Schedule...........................................................................................................................................................7
2.2.1 First Experiment: BG-11 Medium vs. F/2 Part A and Part B Solution................................................................7
2.2.2 Second Experiment: Variations of F/2 Solution.....................................................................................................8
2.2.3 Third Experiment: Nitrate Concentration...............................................................................................................9
2.2.4 Fourth Experiment: Phosphate Concentration....................................................................................................10
2.3 Sample Analysis..................................................................................................................................................................10
2.3.1 Ion Chromatograph...................................................................................................................................................10
2.3.2 Suspended Solids.......................................................................................................................................................11
3.0 Results and Discussion...........................................................................................................................................................14
3.1 First Experiment: BG-11 Medium vs. F/2 Part A and B Solution Results .................................................................14
3.2 Third Experiment: Nitrate Concentration Results........................................................................................................15
3.3 Fourth Experiment: Phosphate Concentration Results...............................................................................................19
3.4 Ion Chromatograph Results .............................................................................................................................................23
3.4.1 Nitrate Concentration Results ................................................................................................................................23
3.4.2 Phosphate Concentration Results..........................................................................................................................25
4.0 Conclusion and Recommendations......................................................................................................................................26
4.1 First Experiment: BG-11 Medium vs. F/2 Part A and B Solution Conclusion...........................................................26
4.2 Third Experiment: Nitrate Concentration Conclusion.................................................................................................26
4.3 Fourth Experiment: Phosphate Concentration Conclusion........................................................................................27
4.4 Ion Chromatograph Conclusion.......................................................................................................................................27
4.5 Conclusion Summary.........................................................................................................................................................27
4.6 Recommendations for Experimental Improvement ....................................................................................................28
5.0 Appendix...................................................................................................................................................................................29
5.1 Raw Data .............................................................................................................................................................................29
5.2 Calculations.........................................................................................................................................................................33
5.2.1 First Experiment Calculations..................................................................................................................................33
5.2.2 Third Experiment Calculations................................................................................................................................33
5.2.3 Fourth Experiment Calculations .............................................................................................................................34
5.3 Gantt Chart..........................................................................................................................................................................36
5.4 Progress Reports ................................................................................................................................................................37
5.5 Literature.............................................................................................................................................................................38
4. 1.0 Introduction
1.1 Literature Review
As mentioned previously, algae is a green, marine-based, photosynthesizing organism that
grows, and lives in, water ("All about Algae Supplements | Precision Nutrition."). Although
algae photosynthesizes, producing energy from sunlight similarly to other land plants, algae is
categorized in a group of its own because as algae does not possess characteristics such as leaves
and roots. Additionally, there are many different types of algae ranging from small, microscopic
single-celled forms (micro-algae), too large to complex multicellular forms (macro-algae) such
as the giant kelp. Furthermore, different types of algae can thrive in both fresh and salt water
conditions.
Because of the diversity in algae and the plenty-fullness of this resource, algae’s viability to
produce crude oil under certain conditions to use as an alternative energy source is becoming
evident. Algae can be grown with relative ease in almost any location. For example, waste
streams can be converted to algae bloom sources without much preparation as they already
contain much of the run off nutrients needed for algae growth.
Researchers, within recent years, have begun to develop several different methods to extract the
algae develop algae biofuels ("Algal Biofuels.”), as well as developing methods to produce the
algae. Specifically, a process known as hydrothermal liquefaction has seen much promise as a
viable means of transforming algae into crude oil or what is commonly termed bio-fuel. The
process mimics the natural processes that convert organic material into crude oil. The process
involves a mixture of algae (20% by weight) and water that is sent down a long tube that holds
the algae at 660 degrees Fahrenheit and 3000 psi for 30 minutes while stirring it (Forbes). By this
process, 100 pounds of algae can yield up to 53 pounds of biofuel (PNNL studies). The oil
produced is of low sulfuric content, which is desired, and is very similar to light, sweet crude,
with a complex mixture of light and heavy compounds, aromatics, phenolics, heterocyclics and
alkanes in the C15 to C22 range (Forbes).
It has been discovered that one of the more effective and efficient types of algae to grow to
harbor biofuels is known as pond scum, or the type of algae that sits on top of ponds (Newman).
This paper will explain the different methods of growing algae and the different parameters that
need to be considered and controlled in order accelerate, and maintain, algae growth.
There are several key elements needed to grow, and maintain the growth, of algae, these include:
temperature, nutrients, light, pH, turbulence, and salinity (Lavens). Finding the balance for each
of these parameters is critical to continue the growth of algae for a long period of time. The
parameter that will be focused on in this paper is the nutrients.
Most micro-algae are strictly photosynthetic; in other words the majority of the growth is
initiated and sustained by light, carbon dioxide, and water. Some algae species are able to
develop in the absence of light by using glucose, or another source of organic carbons, as a food
source, however this experiment will be not be focusing on these algae groups as this type of
algae is more expensive to produce. In addition to sunlight, in order to accelerate algae growth
5. inorganic salts and other minerals such as: nitrogen, phosphorus, iron, and sometimes silicon,
may be introduced as additional food sources for the algae (Algae for Biofuel Production).
1.2 Experimental Objective
The main experimental objective of this experiment is to determine the most effective
combination of nutrients in order to accelerate the algae species Arthrospira Platensis’s
(spirulina) growth. The nutrients experimented with were from different food sources, F/2 Part A
and Part B Solution or BG-11 Medium, as well as nitrate and phosphate additives. The growth
rate was compared to the control algae grown in an incubating tank with no added food sources,
nitrate, or phosphate.
6. 2.0 Materials and Methods
2.1 Experimental Design
The objective of this experiment was to measure the growth and determine which combination of
nutrients provided to the algae would enhance and accelerate growth. As mentioned previously
in the paper, the algal species used in this experiment was the Arthrospira Platensis (spirulina)
due to its sensitivity to various conditions.
One plastic incubator tank was erected in the experimental laboratory room, BC 251. Above this
tank hung a light fixture with lights attached in order to provide uniform light for the incubating
algae, see Figure 1 below.
Figure 1: The 26 Liter Incubating Tank Setup
This incubating tank was the source for all of the experimental algae samples, therefore growth
in this tank began earlier than the rest of the experimental samples. To begin the growth in this
incubating tank, F/2 Part A and Part B Algae Solution was provided once to the algae. If any
water was needed to add to the solution, deionized water was obtained to guarantee no nutrients
in the water would affect the experiment.
Each week, samples would be taken from this incubating tank and these samples would be tested
to determine different results. These results that were to be determined by the conclusion of the
experiment were the type of feed to use, the proper amount of feed, and the proper addition of
nitrate and phosphate in order to accelerate growth. In order to maintain a control sample, each
week a sample would be taken from the incubating tank and would remain untouched. All of
these samples would then be tested to determine the growth rate and the correlating levels of
nitrogen and phosphate. These were determined through suspended solids testing and through the
use of the Ion Chromatograph respectively.
7. 2.2 Experiment Schedule
The experiment can be divided into three smaller experiments, each trying to establish something
different. Table 1 below outlines the order of the occurring sub experiments. For further details,
refer to Gantt Chart in Appendix section.
2.2.1 First Experiment: BG-11 Medium vs. F/2 Part A and Part B Solution
The first experiment that was conducted was the determination of what feed to use, or whether
the feed was unnecessary. There were two types of feed available in the laboratory from the
previous “Algae as a Biofuel” group, therefore BG-11 Medium and F/2 Part A and Part B
solution, as well as a control sample with nothing added, were established, Figure 2 and Figure 3
corresponded to the algae feed respectively.
Figure 2: The BG-11 Medium Figure 3: The F/2 Part A and Part B Solution
A total of seven samples were taken, three to test the BG-11 Medium, three to test the F/2 Part A
and Part B Solution, and one from the incubating tank with nothing added to serve as a control
sample. The control sample taken was only 200 mL of incubated algae tank water. Then, each
sample taken for the BG-11 Medium and F/2 Part A and Part B Solution testing was 1000 mL.
A calculation for the appropriate amount of Medium and Solution needed to add to 1000 mL of
incubating tank water occurred; these calculations and proper amounts can be found in the
Appendix section of this report. In order to assure that the calculated amount of F/2 Part A and
Part B Solution and BG-11 Medium were added to the samples, an Eppendorf Xplorer electronic
pipette was used. Each sample was clearly labeled with a sticker and a name.
Date Test Performed
September 24, 2015 First Experiment: BG-11 Medium vs. F/2 Part
A and Part B Solution
October 1, 2015 Second Experiment: Variations of F/2 Solution
October 8, 2015 Third Experiment: Nitrogen Concentration
November 5, 2015 Fourth Experiment: Phosphate Concentration
Table 1: The Experimental Schedule
8. Next, each sample, except for the control sample, had a portion removed and incubated in small
incubation bottles and stored in the refrigerator in the laboratory room. The rest of the samples
were to remain on the window sill in order to continue growth, Figure 4 represents this.
Figure 4: Samples of Algae Placed on the Window Sill to Continue Growth
However, before these samples were placed on the window sill, small samples were taken from
each 1000 mL sample and placed into a vial in order to test phosphate and nitrogen levels
through the use of the Ion Chromatograph; this procedure can be found under the “Ion
Chromatograph Procedure” section.
Once the samples for the Ion Chromatograph were taken, the remainder of the samples were left
untouched on the window sill for one week. A mark on the flasks or beakers was made at the
1000 mL water level. After a week, both the incubated samples and the window sill samples
underwent a suspended solids testing. The procedure for the suspended solids testing can be
found under the “2.3.2 Suspended Solids” section.
There was a source of experimental error regarding this experiment; in order to begin the algae
growth in the incubation tank, the F/2 Part A and Part B Solution was already used. Therefore,
each sample taken had a small concentration of the F/2 Solution.
2.2.2 Second Experiment: Variations of F/2 Solution
After the F/2 Solution was determined to accelerate algae growth more than the BG-11 Medium
(for these results, refer to the Results section), an experiment using only portions of the F/2
Solution compared to the BG-11 solution was organized.
A sample of F/2 Solution using only the Part A Solution, a sample of F/2 Solution using only the
Part B Solution, and a sample using the BG-11 Medium was created. Each sample was again
labeled using the method in Section 2.2.1. It was established through the suspended solids
analysis this that the most effective algae growth solution was when the F/2 Solution Part A and
9. Part B were combined (refer to the Results section of this report for further details) therefore
each experiment from this point involved only F/2 Solution, both Parts A and B.
2.2.3 Third Experiment: Nitrate Concentration
The third experiment was to determine the best combination of varying levels of added nitrate
and varying levels of F/2 Solution. In order to add additional nitrate to the solution, Sodium
Nitrate was chosen.
To better display and organize this experiment, a table was created.
No Additional
Nitrate
30 mg/L of
Additional Nitrate
60 mg/L of
Additional Nitrate
No F/2 Solution 1 sample 1 sample 1 sample
Low Level of F/2 Solution 1 sample 1 sample 1 sample
Baseline Level of F/2 Solution 1 sample 1 sample 1 sample
High Level of F/2 Solution 1 sample 1 sample 1 sample
Table 2: Level of F/2 Solution and Concentration of Additional Nitrate Samples
Each sample was clearly labeled, refer to Section 2.2.1 on the procedure.
To guarantee that the correct concentration of nitrate was added to each sample, a calculation
occurred; this calculation can be referred to in the Appendix Section of the report. This
calculated amount of Sodium Nitrate was poured out onto a Fisherbrand Low Form Aluminum
Flute 42 mL Weighing Disk and then measured on a Denver Instrument Company mass balance
Scale to measure the proper weight. An additional sample from the incubating tub was also
taken.
To create each sample, the incubating tub water was stirred to assure no algae settling. Then, 300
mL of incubating tub water containing the algae was collected. Then, 1200 mL of deionized
water was measured and combined with the 300 mL of incubating tub water in a 2000 mL
Volumetric Flask in order to create a 1500 mL sample. From this, 500 mL was poured into a
plastic bottle and incubated for a week and the 1000 mL remainder of the sample was placed in
either a 1000 mL Erlenmeyer flask or a 1000 mL Beaker and set aside to continue growth for a
week.
The order for the creation of the samples was to begin creating the “No F/2 Solution” and
advanced down the column, refer to Table 2. Then, the “30 mg/L of Additional Nitrate” solution
was created and more samples were created by advancing down the column, again refer to Table
2. The same procedure occurred for the “60 mg/L of Additional Nitrate” column.
Then, following the First and Second Experiments, small samples were taken from each sample
created for the Ion Chromatograph testing and after the week passed, a suspended solids test was
run with the incubated samples as well as the samples that were allowed to grow on the window
sill.
10. 2.2.4 Fourth Experiment: Phosphate Concentration
The Fourth experiment emulated the Third Experiment, with the main exception being that the
objective was to test the most effective level of phosphate with varying levels of F/2 Solution.
This was done by using Potassium Phosphate as the additive. The below table displays the
varying levels of phosphate and F/2 Solution added.
No Additional
Phosphate
5 mg/L of Additional
Phosphate
10 mg/L of
Additional Phosphate
No F/2 Solution 1 sample 1 sample 1 sample
Low Level of F/2
Solution
1 sample 1 sample 1 sample
Baseline Level of F/2
Solution
1 sample 1 sample 1 sample
High Level of F/2
Solution
1 sample 1 sample 1 sample
Table 3: Level of F/2 Solutions and Concentration of Additional Phosphate Samples
Similarly to the Third Experiment, a calculation occurred to determine the correct concentration
of phosphate was added to each sample; this calculation can be referred to in the Appendix
Section of the report. To understand the labeling, refer to Section 2.2.1, then to understand the
procedure used to measure these concentrations, refer to Section 2.2.3 of the report. An
additional sample from the incubating tub was also taken.
All the volumes of all samples in this experiment are identical to the volumes used in the Third
Experiment, so refer to Section 2.2.3 for volumes. Furthermore, the order of sample creations
was identical to the Third Experiment (with the exception that the Fourth Experiment began with
the column “No Additional Phosphate” instead of “No Additional Nitrate” and then proceeded to
“5 mg/L of Additional Phosphate” instead of “30 mg/L of Additional Nitrate). To review this
procedure, refer to Section 2.2.3 of the report.
Finally, following the procedure of all the previous experiments, after a week of incubation and
growth, an Ion Chromatograph test and a suspended solids test occurred.
2.3 Sample Analysis
Once all of the above data was created by way of the four experiments, two methods of analysis
were used to generate results and determine which combinations of F/2 Solution, nitrate, and
phosphate. These two tests were the solids suspended testing and the Ion Chromatograph testing.
The procedures are seen below.
2.3.1 Ion Chromatograph
The first analysis that occurred was the Ion Chromatograph analysis. In order to prepare for this,
a syringe, a syringe filter, and test vials were needed.
From each sample that used an Ion Chromatograph analysis, the syringe needed to be flushed to
guarantee that no contaminates would affect the results. The syringe was flushed with each twice
11. and these two flushing’s were wasted into a 100 mL beaker. Once the syringe was properly
flushed for the proper sample, another small portion of the sample was pulled up into the syringe.
Then, a filter was placed onto the end of the syringe and the small portion of the sample
collected was pushed through the filter, into a small vial corresponding to the Ion
Chromatograph. Each vial was properly labeled. The syringe was flushed twice, a new filter, and
new vials were obtained used for each new sample of every experiment.
Once this was completed, the vial labeling was then input to the computer that controlled the Ion
Chromatograph; each vial needed to be inserted in the Ion Chromatograph with the proper
number labeling for further analysis once the Ion Chromatograph produced results.
Figure 5: The Ion Chromatograph
The results produced from the Ion Chromatograph reflected the concentrations of the nitrate or
phosphate that remained after the algae consumed the nutrients. These results can be found in
Section 3.0 Results and Discussion.
2.3.2 Suspended Solids
The objective of this analysis was to compare the growth of the incubated samples of algae from
each Experiment to the samples from each Experiment that were allowed left on the window sill
for the week, as well as which levels of additional nitrate, phosphate, and level of F/2 Solution
allowed the most growth.
After allowing the samples to rest for a week, either in incubation in the refrigerator or on the
window sill, these samples were taken in the laboratory room. The samples that were on the
window sill needed to be filled with deionized water to the 1000 mL level mark made on the
flasks or beakers in the First Experiment section.
Sheets of Whatman 934-AH 90 Diameter Glass Microfibre Filter paper were then labeled to
reflect the labeling on each sample flask or beaker. Then, this filter paper was weighed on the
Denver Instrument Company Mass Balance Scale and its weight was recorded on a sheet of
paper in grams with the correct sample labeling. Next to this, the volume of the sample was
12. recorded as well. Once these values were recorded, the paper was inserted into a filter funnel.
The filter funnel was then inserted into a 1000 mL Erlenmeyer Vacuum Flask which was
attached to a Vacuum Pump by way of a rubber hose; this set up can be seen in Figure 6 below.
Figure 6: Suspended Solids Testing Setup
Next, the filter paper was wet with deionized water and the vacuum pump was turned on. Once
the pump was running, the flask or beaker sample that corresponded to the labeled filter paper
would be poured through the filter funnel; deionized water was used to wash the flask or beaker
the sample was being poured from to assure that all of the sample was run through the filter
funnel.
Once all of the sample had been poured out of its original flask or beaker and had been run
through the filter funnel, the vacuum pump was turned off mand the filter paper was removed
and placed into a Fisher Scientific Isotemp Oven to dry for an hour, see Figure 7. The 1000 mL
Erlenmeyer flask was then drained. This procedure occurred for each sample.
Figure 7: The Fisher Scientific Isotemp Oven
After each sample had been in the oven for an hour, the filter paper was removed from the oven
and then weighed in grams again, see Figure 8 for the dried suspended solids filter paper. This
13. new weight of the filter paper was again recorded corresponding with the correct sample
labeling.
Figure 8: Samples of Dried Filter Paper from the Suspended Solids Test
To find the suspended solids concentration, this original weight of the filter paper was subtracted
from the new weight of the filter paper for each sample. This value in grams was then converted
to milligrams and divided by the recorded sample volume in liters.
The results produced from the suspended solids testing reflect which combination of phosphate
and F/2 Solution, and nitrate and F/2 Solution generated the most growth; the filter paper with
the highest suspended solids value meant the combination of feed and nutrients experienced the
most growth.
All these values were recorded in Tables that can be found in section 5.0 Appendix and analyzed
further. The further analysis can be found in Section 3.0 Results and Discussion.
14. 3.0 Results and Discussion
This section displays all the data collected throughout the project with the objective being to
determine the optimum nutrient concentrations to us in order to accelerate algae growth. The
nutrients included the algae food, BG-11 Medium and F/2 Part A and Part B Solution and
additional nutrients, nitrate and phosphate, as mentioned in Section 2.0 of the report. All growth
was based on a log distribution between the results of the initial week of sample testing and the
last week of sample testing. In other words, this can be represented as:
𝐿𝑜𝑔(
𝐿𝑎𝑠𝑡 𝑊𝑒𝑒𝑘 𝑜𝑓 𝑆𝑢𝑠𝑝𝑒𝑛𝑑𝑒𝑑 𝑆𝑜𝑙𝑖𝑑𝑠 𝐺𝑟𝑜𝑤𝑡ℎ
𝐼𝑛𝑖𝑡𝑖𝑎𝑙 𝑊𝑒𝑒𝑘 𝑜𝑓 𝑆𝑢𝑠𝑝𝑒𝑛𝑑𝑒𝑑 𝑆𝑜𝑙𝑖𝑑𝑠 𝐺𝑟𝑜𝑤𝑡ℎ
)
This equation will represent the values in all Tables and graphs with the exception of the values
in Table 4 and Figure 9.
3.1 First Experiment: BG-11 Medium vs. F/2 Part A and B Solution Results
As seen in Section 2.0, the first experiment performed was a determination of which algae food
to use to accelerate growth, either BG-11 Medium or F/2 Part A and B Solution. Table 4 below
presents the data generated from the suspended solids tests which compares the growth in algae
when fed these two food. This table also presents the average suspended solids growth based on
the different foods as well.
17-Sep Suspended Solids
(mg/L)
Total Food
Added (𝝁L)
BG-11 #1 Sample 12.5 3359
BG-11 #2 Sample 14.125 3359
BG-11 #3 Sample 11.625 3359
F/2 #1 Sample 23.75 6718
F/2 #2 Sample 19.5 6718
F/2 #3 Sample 36.25 6718
BG-11 Average 12.75
F/2 Average 26.5
Table 4: Suspended Solids Weight and Total Food Added for the Flask Samples
15. Figure 9 below is the graph of the average suspended solids growth based on food.
Figure 9: BG-11 Medium vs. F/2 Solution Suspended Solids Growth
As seen through Table 4 and Figure 9, F/2 Part A and B Solution is the preferred algae food
source as it generated the largest suspended solids value, or the most growth, therefore the
experiment proceeded using only F/2 Solution as the Algae feed. For more discussion on this,
refer to Section 4.1.
3.2 Third Experiment: Nitrate Concentration Results
The third experiment was to test various levels of F/2 Part A and B Solution to determine the F/2
Solution concentration as well as the concentration of additional nitrate that would generate the
optimal algae growth, as referenced in Section 2.0 of the report. Table 5 displays the suspended
solids growth data, dependent on the varying amount of F/2 Part A and B Solution and the
additional nitrate added. Table 5 below displays this data, and this table and the below figures
reference the log equation.
Figure 10 demonstrated the logarithmic graph of the suspended solids test with no nitrate added
and varying levels of F/2 Solution added.
0 N 30 N 60 N
No F2 0.12 0.15108 0.25103
97 μl/L F2 0.275528 0.284417 0.268115
194 μl/L F2 0.390388 0.474735 0.390006
388 μl/L F2 0.424033 0.498511 0.338535
Table 5: Logarithmic Suspended Solids Growth Based on Varying F/2 Solution and Nitrate Concentrations
0
5
10
15
20
25
30
35
40
BG-11 F2
Suspended
Solids
(mg/L)
Algae Growth for RecommendedFeed
Concentration
16. Figures 10, 11, and 12 are graphical representations of the data presented in Table 5.
Figure 10: Logarithmic Suspended Solids Growth with Varying Levels of F/2 Solution and No Additional Nitrate
Figure 11: Logarithmic Suspended Solids Growth with Varying Levels of F/2 Solution and 30 mg/L of Nitrate Added
0
0.1
0.2
0.3
0.4
0.5
No F2 97 μl/L F2 194 μl/L F2 388 μl/L F2
Log
Growth
wk3/wk1
Suspended Solids with No Nitrate
0
0.1
0.2
0.3
0.4
0.5
0.6
No F2 97 μl/L F2 194 μl/L F2 388 μl/L F2
Log
Growth
wk3/wk1
Suspended Solids with 30mg/L Nitrate
17. Figure 12: Logarithmic Suspended Solids Growth with Varying Levels of F/2 Solution and 60 mg/L of Nitrate Added
It can be seen from the above figures that adding no F/2 Solution resulted in very poor algae
growth as the columns with “No F2” have small logarithmic suspended solids values. It can also
by observed by the above figures that the optimal amount of added F/2 Solution for any amount
of nitrate added was determined to be 194 𝜇L as this value experienced the largest logarithmic
growth in all the figures. Further discussion of this can be found in Section 4.2.
However, to confirm these two observations, Table 6 presents the data for logarithmic suspended
solids growth dependent on 194 𝜇L, 97 𝜇L, and 388 𝜇L of F/2 Solution added and varying
amounts of Nitrate added.
Suspended solids based on F2 amounts with various Nitrate
Amounts
194 𝝁L of F2 97 𝝁L of F2 388 𝝁L of F2
0 mg/L Nitrate 0.390388 0.275528 0.424033
30 mg/L Nitrate 0.474735 0.284417 0.498511
60 mg/L Nitrate 0.390006 0.268115 0.338535
Table 6: Logarithmic Suspended Solids Growth Based on Varying F/2 Solution (No Control Sample) and Nitrate
Concentrations
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
No F2 97 μl/L F2 194 μl/L F2 388 μl/L F2
Log Growth
wk3/wk1
Suspended Solids with 60mg/L Nitrate
18. Figures 13, 14 and 15 correspond with Table 6. These figures graphically display the different
amounts of nitrate would give optimal growth based on suspended solids at different F/2
amounts.
Figure 13: Logarithmic Suspended Solids Growth with 194 𝜇L of F/2 Solution and Varying Levels of Nitrate
Concentrations
Figure 14: Logarithmic Suspended Solids Growth with 97 𝜇L of F/2 Solution and Varying Levels of Nitrate
Concentrations
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
0 mg/L Nitrate 30 mg/L Nitrtate 60 mg/L Nitrate
Log Growth
wk3/wk1
Effect of Nitrate
194 μL F2
0.255
0.26
0.265
0.27
0.275
0.28
0.285
0.29
0 mg/L Nitrate 30 mg/L Nitrtate 60 mg/L Nitrate
Log Growth
wk3/wk1
Effect of Nitrate
97 μL F2
19. Figure 15: Logarithmic Suspended Solids Growth with 388 𝜇L of F/2 Solution and Varying Levels of Nitrate
Concentrations
After analysis of all the above figures, it can be concluded that in terms of nitrate and F/2
concentrations, the combination of 194 𝜇L of F/2 Solution added with 30 mg/L of nitrate is the
most optimal concentration. To further confirm that this was the optimal level of F/2 Solution, as
well as determine the optimal amount of phosphate needed, the fourth experiment, refer to
Section 2.2.4, was analyzed.
3.3 Fourth Experiment: Phosphate Concentration Results
The fourth experiment was to test various levels of F/2 Part A and B Solution and phosphate
added to determine the ideal combination of both to generate the optimal algae growth, as
referenced in Section 2.0 of the report. Table 7 displays the suspended solids growth data,
dependent on the varying amount of F/2 Part A and B Solution and the amount of additional
phosphate. This table and the below figures reference the log equation.
Suspended solids growth based on Phosphate and F2
0 mg/L of P 5 mg/L of P 10 mg/L of P
No F2 0.169887 0.063549 0.128765
97 μl/L F2 0.175263 0.13099 0.106089
194 μl/L F2 0.438353 0.172933 0.083134
388 μl/L F2 0.036517 0.083546 -0.0202
Table 7: Logarithmic Suspended Solids Growth Based on Varying F/2 Solution and Phosphate Concentrations
0
0.1
0.2
0.3
0.4
0.5
0.6
0 mg/L Nitrate 30 mg/L Nitrtate 60 mg/L Nitrate
Log Growth
wk3/wk1
Effect of Nitrate
388 μL F2
20. Figures 16, 17, and 18 are graph representations of the information displayed in Table 7.
.
Figure 16: Logarithmic Suspended Solids Growth with Varying Levels of F/2 Solution and No Additional Phosphate
Figure 17: Logarithmic Suspended Solids Growth with Varying Levels of F/2 Solution and 5 mg/L of Phosphate
0
0.1
0.2
0.3
0.4
0.5
No F2 97 μl/L F2 194 μl/L F2 388 μl/L F2
Log Growth
wk2/wk1
Suspended Solids with No Phosphate
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0.2
No F2 97 μl/L F2 194 μl/L F2 388 μl/L F2
Log Growth
wk2/wk1
Suspended Solids with 5mg/LPhosphate
21. Figure 18: Logarithmic Suspended Solids Growth with Varying Levels of F/2 Solution and 10 mg/L of Phosphate
Referencing this data above, the determination can be made that for most concentration of
phosphate added, not adding F/2 Solution did not assist algae growth. However, again a
confirmation needed to occur, therefore Table 8 below displays data from 97 𝜇L, 194 𝜇L, and
388 𝜇L of F/2 Solution added compared to the various concentrations of phosphate shown above.
Suspended solids based on F2 amounts with various Phosphate
Amounts
194 𝝁L 97 𝝁L 388 𝝁L
0 mg/L Phosphate 0.438352713 0.17526324 0.036516834
5 mg/L Phosphate 0.17293274 0.17293274 0.083546051
10 mg/L Phosphate 0.083133551 0.08313355 -0.02020339
Table 8: Logarithmic Suspended Solids Growth Based on Varying F/2 Solution (No Control Sample) and Phosphate
Concentrations
-0.04
-0.02
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
No F2 97 μl/L F2 194 μl/L F2 388 μl/L F2
Log Growth
wk2/wk1
Suspended Solids with 10mg/LPhosphate
22. Figures 19, 20 and 21 correspond with Table 8; these are graphs to show which amount of
Phosphate would give optimal growth based on suspended solids at different F2 amounts.
Figure 19: Logarithmic Suspended Solids Growth with 194 𝜇L of F/2 Solution and Varying Phosphate Concentrations
Figure 10: Logarithmic Suspended Solids Growth with 97 𝜇L of F/2 Solution and Varying Phosphate Concentrations
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0.2
0 ml Phosphate 5 ml Phosphate 10 ml Phosphate
Log Growth
wk2/wk1
Effect of Phosphate
97 μL F2
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
0 ml Phosphate 5 ml Phosphate 10 ml Phosphate
Log Growth
wk2/wk1
Effect of Phosphate
194 μL F2
23. Figure 11: Logarithmic Suspended Solids Growth with 388 𝜇L of F/2 Solution and Varying Phosphate Concentrations
By observing the above figures, it can be verified that the most successful combination of F/2
Solution and added phosphate is 194 𝜇L and 0 mg/L, respectively. Further discussion of this can
be referenced in Section 4.3.
3.4 Ion Chromatograph Results
The following tables and graphs present the data collected from the Ion Chromatograph; the Ion
Chromatograph tested nitrate and phosphate levels that was consumed by the algae. Similar to
Section 3.1 through 3.3, these graphs compare the initial week of sample to the final week of
sample.
3.4.1 Nitrate Concentration Results
Figures 22 and 23 on the following page represent the amount of nitrate that the algae consumed.
These figures objectives are to prove a relationship between nitrate consumed and algae growth.
Please see appendix for the Ion Chromatograph data used for these graphs, highlighted in yellow.
-0.04
-0.02
0
0.02
0.04
0.06
0.08
0.1
0 ml Phosphate 5 ml Phosphate 10 ml Phosphate
Log Growth
wk2/wk1
Effect of Phosphate
388 μL F2
24. Figure 12: Suspended Solids vs. Nitrate Consumed Linear Relationship
Figure 13: Suspended Solids vs. Nitrate Consumed for Various F/2 Solution Levels
y = 3.9174x
R² = 0.0394
0
10
20
30
40
50
60
70
80
0 5 10 15 20
Suspended Soilds
(mg/L)
Nitrate Consumed (mg/L)
Suspended Solids vs. Nitrate Consumed
(wk3 vs wk1)
0
10
20
30
40
50
60
70
0 5 10 15 20
Suspended Soilds
(mg/L)
Nitrate Consumed (mg/L)
Suspendid Solids vs. Nitrate Consumed
(wk3 vs wk1)
No F2
Baseline F2
Low F2
High F2
25. 3.4.2 Phosphate Concentration Results
Figures 24 and 25 represent the amount of phosphate that the algae consumed. These figures
objectives are to prove a relationship between nitrate consumed and algae growth. Please see
appendix for the Ion Chromatograph data used for these graphs, highlighted in yellow.
Figure 14: Suspended Solids vs. Phosphate Consumed Linear Relationship
Figure 15: Suspended Solids vs. Phosphate Consumed for Various F/2 Solution Levels
y = 631.17x
R² = -0.657
-5
0
5
10
15
20
25
30
35
40
45
0 0.005 0.01 0.015 0.02 0.025 0.03
Suspended Soilds
(mg/L)
Phosphate Consumed (mg/L)
Phosphate Consumed vs Suspendid Solids
(wk2 vs wk1)
-5
0
5
10
15
20
25
30
35
40
45
0 0.005 0.01 0.015 0.02 0.025 0.03
Suspended Soilds
(mg/L)
Phosphate Consumed (mg/L)
Phosphate Consumed vs Suspendid Solids
(wk3 vs wk1)
No F2
Baseline F2
Low F2
High F2
26. 4.0 Conclusion and Recommendations
4.1 First Experiment: BG-11 Medium vs. F/2 Part A and B Solution Conclusion
The results indicated that over the course of three weeks, the F/2 Solution Parts A and B yielded
an average suspended solids of 26.5 mg/L. This suspended solids value was then compared to
suspended solids value produced by the BG-11 Medium, which was 12.75 mg/L for BG-11. The
difference between these two values is determined to be 13.75 mg; therefore indicating that the
F/2 Solution generated more algae growth. Furthermore, the trend was linear through the
progression of the time that was analyzed, meaning that the amount of solids, or algae, in the
water increased each successive for the F/2 Solution.
With the combined knowledge of both these results, it can be concluded that F/2 Solution had a
greater impact on the amount of suspended solids generated with time than the BG-11 Medium,
meaning that the F/2 is the optimal algae feed to use.
However, there is an experimental error associated with this. This trend generated may not
irrevocably conclusive because the amount of F/2 Solution was double that added of the BG-11
corresponding samples. This is due to the fact that the F/2 Solution has a Part A and Part B
Solution, therefore meaning that 194 𝜇L of Part A and 195 𝜇L of Part B was added to samples of
F/2 making a total of 388 𝜇L added to F/2 Solution samples whereas only 194 𝜇L was added to
the BG-11 Medium samples.
4.2 Third Experiment: Nitrate Concentration Conclusion
The first determination made in this experiment was by referring to Figures 10 through 12. These
figures indicated that the baseline amount of F/2 Solution consistently produced greater log
growth between week three and week one than any other amount of F/2 Solution added.
Therefore, overall the baseline amount of F/2 Solution, or 194 𝜇L of F/2 Solution, was
determined to be the optimal level of F/2 Solution for algae growth referring solely to this data in
this experiment.
Furthermore, the second determination made was the concentration of nitrate needed for optimal
algae growth. As the nitrate concentration increased, along with the increase of F/2 Solution, the
trend was that more algae was produced. However, at some point this trend reversed and by
adding more F/2 Solution and more nitrate the amount of algae produced decreased.
One reason for this occurring may be due to the oversaturation of nitrates in solution that begins
to hamper algae growth This problem may have also been compounded by the fact that it is
assumed that F/2 algae food already contains a certain percentage of nitrates as its makeup; as
more nutrients were provided more algae grew at a faster rate. At some point though, the
exponential growth of algae consumed all of the nutrients. Once this occurred algae began to
die.
As expressed earlier, it can be concluded that, using these experimental values alone, the optimal
level of F/2 Solution added would be 194 𝜇L. However the data suggests a high likelihood that
the optimum F/2 Solution level may be somewhere in between 194 𝜇L and 388 𝜇L.
27. In terms of the optimum levels of nitrates added, 30 mg/L consistently showed the highest log
growth when compared across varying levels of F/2 Solution. Higher levels of added nitrate
consistently recorded lower amounts of log growth, confirming the oversaturation hypothesis in
the preceding paragraph.
4.3 Fourth Experiment: Phosphate Concentration Conclusion
For effects of phosphate analysis, the trends were similar to that of the nitrates and hence some
of the conclusions made from the analysis of the effects of nitrates also apply to phosphates.
Again the greatest log growth was recorded by the baseline level (194 𝜇L) of F/2 Solution for the
two smallest concentrations of phosphates added.
Furthermore, it can be observed that the more phosphate added to the solution, the less growth
the algae experienced. Similarly to oversaturating the solution with nitrate, the solution may have
been oversaturated with phosphates which negatively impacted algae growth. As mentioned
previously it is assumed the F2 food already contains the certain percentage of phosphate thus
may be contributing to oversaturation of nutrients. However, the oversaturation concentration of
phosphate was much lower than the oversaturation concentration of nitrate. This may be
explained due to the fact that phosphates are less commonly found in natural environments,
meaning algae needs less phosphate to accelerate growth when compared to nitrate.
Unfortunately, the results of the optimal concentration of phosphate were inclusive since no
noticeable trend could be determined from the results. However, it may be noteworthy to
mention that the highest growth level was recorded by the sample with no additional phosphate
added at 194 𝜇L of F/2 Solution (the level of F/2 Solution deemed optimal by both the third and
fourth experiment). Furthermore, the lowest growth was consistently recorded by the highest
level of phosphate concentration despite the different levels of F/2 Solution examined.
4.4 Ion Chromatograph Conclusion
The correlation between nutrients consumed and algae grown seemed to correlate with the
conclusions made previously. However, there was no linear correlation to the nutrients consumed
and suspended solids growth. In general, the nitrate consumed showed a positive relationship
with the amount of suspended solids recorded. In contrast, the consumption of phosphate
generally showed a negative correlation when compared to the suspended solids value. Therefore
there seems to be some adverse effects of excess phosphate on algae growth, and the amount
necessary for this to occur happens is relatively small.
Moreover, the regression constant of the trend line was generated by the phosphate concentration
variation graph was very low, indicating that the results were relatively scattered and
reproducibility of the experiment is questionable.
4.5 Conclusion Summary
The optimum algae food was found to be F/2 Solution both Parts A and B at a concentration of
194 𝜇L. The optimum nitrate level used in combination with F/2 Solution was found to be 30
28. mg/L and the optimum phosphate concentration was inconclusive although results suggest that
no additional phosphate may be required apart from that already present in F/2 Solution.
4.6 Recommendations for Experimental Improvement
1. The experiment should be redone to establish F/2 Solution as the better medium for algae
growth; the portions of Parts A and B of F/2 Solution should be halved and the
experiment repeated, or the amount of BG-11 added per sample should be increased.
2. A more accurate optimum F/2 level should be determined by taking smaller increments
of F/2 Solution levels starting at 194 𝜇L and ending at 388 𝜇L.
3. An analysis of the composition of F/2 Solution should be conducted more thoroughly in
order to establish the percentage nitrates and phosphates used in the Solution. This would
help establish more accurate optimal concentrations of nitrate and phosphate.
4. Overtime the pH of the solution samples should be recorded to determine correlation
between nutrient concentrations and pH levels. This is necessary because pH levels may
have considerable impact on algae growth. One means of acidification of water bodies is
through the production of ammonia, and hence the conversion of nitrates to ammonia is
possible, and hence acidification of the samples may be possible as a consequence.
5. Future experiments should consider storing samples with a constant light source as light
intensity directly affects algae growth. This is important because over the course of the
experiment, the amount of light slowly decreased.
29. 5.0 Appendix
5.1 Raw Data
BG-11 vs F2 Suspended Solids Weight in the Incubated Samples
24-Sep
Volume
Used
(mL)
Weight
of Filter
Paper (g)
Weight of Dried
FilterPaperwith
Algae (g)
Weight
Difference
(g)
Suspended
Solids
(g/mL)
Suspended
Solids
(mg/mL)
BG-11 #1 Sample 80 0.4062 0.4055 -0.0007 -8.75E-06 -0.00875
BG-11 #2 Sample 60 0.4017 0.4013 -0.0004 -6.667E-06 -0.00666667
BG-11 #3 Sample 80 0.4078 0.4076 -0.0002 -2.5E-06 -0.0025
F2 #1 Sample 80 0.4032 0.4029 -0.0003 -3.75E-06 -0.00375
F2 #2 Sample 80 0.4042 0.4036 -0.0006 -7.5E-06 -0.0075
F2 #3 Sample 73 0.4006 0.4009 0.0003 4.1096E-06 0.004109589
BF-11 vs F2 Suspended Solids Weight in the Flask Samples
1-Oct
Volume
Used
(mL)
Weight
of Filter
Paper (g)
Weight of Dried
FilterPaperwith
Algae (g)
Weight
Difference
(g)
Suspended
Solids
(g/mL)
Suspended
Solids
(mg/mL)
BG-11 #1 Sample 800 0.4067 0.4167 0.01 0.0000125 0.0125
BG-11 #2 Sample 800 0.4041 0.4154 0.0113 1.4125E-05 0.014125
BG-11 #3 Sample 800 0.4035 0.4128 0.0093 1.1625E-05 0.011625
F2 #1 Sample 800 0.4014 0.4204 0.019 0.00002375 0.02375
F2 #2 Sample 800 0.4046 0.4202 0.0156 0.0000195 0.0195
F2 #3 Sample 800 0.4018 0.4308 0.029 0.00003625 0.03625
BG-11 vs F2 Suspended Solids in the Tank Samples
Volume
Used
(mL)
Weight of
Filter Paper
(g)
Weight of Dried
FilterPaperwith
Algae (g)
Weight
Difference
(g)
Suspended
Solids
(g/mL)
Suspended
Solids (mg/mL)
Tank Sample 200 0.4071 0.4188 0.0117 5.85E-05 0.0585
31. Phosphate Suspended Solids Weight of Samples dated 11/19
Volume
Used
(mL)
Weight of
Filter Paper
(g)
Weight of Dried
FilterPaperwith
Algae (g)
Weight
Difference
(g)
Suspended
Solids
(g/mL)
Suspended
Solids (mg/L)
0P BL F2 500 0.3980 0.4312 0.0332 0.0000664 66.4
0P H F2 500 0.4001 0.4249 0.0248 0.0000496 49.6
0P L F2 500 0.3995 0.4257 0.0262 0.0000524 52.4
0P N F2 500 0.3990 0.4268 0.0278 0.0000556 55.6
10P 0 F2 500 0.4007 0.4311 0.0304 0.0000608 60.8
10P BL F2 500 0.3992 0.4279 0.0287 0.0000574 57.4
10P H F2 500 0.3987 0.4197 0.021 0.000042 42
10P L F2 500 0.3969 0.4232 0.0263 5.26E-05 52.6
5P 0 F2 500 0.3996 0.4187 0.0191 0.0000382 38.2
5P BL F2 500 0.3990 0.4264 0.0274 0.0000548 54.8
5P H F2 500 0.3960 0.4200 0.024 4.8E-05 48
5P L F2 500 0.3961 0.4226 0.0265 5.3E-05 53
TUB 500 0.3951 0.5262 0.1311 0.0002622 262.2
Nitrate Suspended Solids Weight of Samples dated 10/8
Volume
Used
(mL)
Weight of
FilterPaper
(g)
Weight of Dried
FilterPaperwith
Algae (g)
Weight
Difference
(g)
Suspended
Solids
(g/mL)
Suspended
Solids (mg/L)
BL F2 0N 500 0.4021 0.4149 0.0128 0.0000256 25.6
BL F2 30N 500 0.4120 0.4208 0.0088 1.76E-05 17.6
BL F2 60N 500 0.3991 0.4110 0.0119 2.38E-05 23.8
H F2 0N 500 0.4005 0.4124 0.0119 2.38E-05 23.8
H F2 30 N 500 0.4058 0.4162 0.0104 0.0000208 20.8
H F2 60 N 500 0.4000 0.4131 0.0131 0.0000262 26.2
L F2 0N 500 0.4078 0.4241 0.0163 0.0000326 32.6
L F2 30N 500 0.4040 0.4148 0.0108 0.0000216 21.6
L F2 60N 500 0.3992 0.4106 0.0114 0.0000228 22.8
N F2 0 N 500 0.4044 0.4180 0.0136 0.0000272 27.2
N F2 30N 500 0.4025 0.4171 0.0146 0.0000292 29.2
N F2 60N 500 0.4054 0.4175 0.0121 0.0000242 24.2
TUB 500 0.3959 0.4495 0.0536 0.0001072 107.2
32. Nitrate Suspended Solids Weight of Samples dated 10/15
Volume
Used
(mL)
Weight of
Filter
Paper (g)
Weight of Dried
FilterPaperwith
Algae (g)
Weight
Difference
(g)
Suspended
Solids (g/mL)
Suspended
Solids (mg/L)
BL F2 0N 500 0.4038 0.4162 0.0124 0.0000248 24.8
BL F2 30N 500 0.4111 0.4245 0.0134 2.68E-05 26.8
BL F2 60N 500 0.4026 0.4200 0.0174 3.48E-05 34.8
H F2 0 N 500 0.4098 0.4220 0.0122 0.0000244 24.4
H F2 30N 500 0.4096 0.4228 0.0132 0.0000264 26.4
H F2 60N 500 0.4078 0.4279 0.0201 0.0000402 40.2
L F2 0N 500 0.4050 0.4189 0.0139 2.78E-05 27.8
L F2 30N 500 0.4069 0.4232 0.0163 3.26E-05 32.6
L F2 60N 500 0.4106 0.4320 0.0214 4.28E-05 42.8
N F2 0 N 500 0.4094 0.4299 0.0205 0.000041 41
N F2 30 N 500 0.4076 0.4289 0.0213 0.0000426 42.6
N F2 60N 500 0.4026 0.4206 0.018 3.6E-05 36
TUB 500 0.4084 0.4230 0.0146 0.0000292 29.2
Table 1: Week of October 29, 2015 Suspended Solids for Nitrate Variation Testing
Sample
Name
Volume
(mL)
Volume
(L)
Initial Weight
of Filter
Paper (g)
Initial Weight
of Filter
Paper (mg)
Weight of
Algae & Filter
Paper (g)
Weight of
Algae & Filter
Paper (mg)
Suspended
Solids
(mg/L)
BL F2 0N 494 0.494 0.3984 398.4 0.4285 428.5 60.931
BL F2 30N 494 0.494 0.3975 397.5 0.437 437 79.960
BL F2 60N 494 0.494 0.3955 395.5 0.4377 437.7 85.425
H F2 0N 494 0.494 0.3977 397.7 0.4297 429.7 64.777
H F2 30N 494 0.494 0.3972 397.2 0.4383 438.3 83.198
H F2 60N 494 0.494 0.3957 395.7 0.439 439 87.652
L F2 0N 494 0.494 0.3994 399.4 0.4253 425.3 52.429
L F2 30N 494 0.494 0.3975 397.5 0.4285 428.5 62.753
L F2 60N 494 0.494 0.3969 396.9 0.4361 436.1 79.352
N F2 0N 494 0.494 0.3992 399.2 0.4259 425.9 54.049
N F2 30N 494 0.494 0.3989 398.9 0.4287 428.7 60.324
N F2 60N 494 0.494 0.3959 395.9 0.4276 427.6 64.170
Tub 494 0.494 0.3962 396.2 0.4696 469.6 148.583
33. 5.2 Calculations
5.2.1 First Experiment Calculations
5.2.1.1 Determination of Amount of F/2 Part A Solution Needed
Part A requires 0.005 L of Solution to every 38.7 L of water.
The sample created was 1 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1 𝐿
𝑥 = 129 𝜇𝐿
5.2.1.2 Determination of Amount of F/2 Part B Solution Needed
Part B requires 0.005 L of Solution to every 38.7 L of water.
The sample created was 1 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1 𝐿
𝑥 = 129 𝜇𝐿
5.2.1.3 Determination of Amount of BG-11 Medium Needed
No indication of ratio of Medium to water, therefore it was assumed to be the same ratio
as the F/2 Part A and F/2 Part B Solution.
The sample created was 1 L.
0.005 𝐿
38.7 𝐿
=
𝑥
26 𝐿
𝑥 = 129 𝜇𝐿
5.2.2 Third Experiment Calculations
5.2.2.1 Determination of Low Level F/2 Solution (both for Part A and for Part B)
Both Part A and Part B require 0.005 L of Solution to every 38.7 L of water.
The sample required was 1.5 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1.5 𝐿
𝑥 = 194 𝜇𝐿
Low Level of F/2 Solution was determined to only need half of this amount
(0.5) ∗ (194 𝜇𝐿) = 𝑥
𝑥 = 97 𝜇𝐿
5.2.2.2 Determination of Baseline F/2 Solution (both for Part A and for Part B)
Part A requires 0.005 L of Solution needed to 38.7 L of water.
The sample created was 1.5 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1 𝐿
𝑥 = 129 𝜇𝐿
5.2.2.3 Determination of High F/2 Solution (both for Part A and for Part B)
Both Part A and Part B require 0.005 L of Solution to every 38.7 L of water.
34. The sample created was 1.5 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1.5 𝐿
𝑥 = 194 𝜇𝐿
Low Level of F/2 Solution was determined to need double this amount
(2)∗ (194 𝜇𝐿) = 𝑥
𝑥 = 388 𝜇𝐿
5.2.2.4 Determination of 30 mg/L Additional Nitrate
Molar mass of Sodium Nitrate: 84.99 g/mol
Molar mass of Nitrate: 62 g/mol
The sample created was a 2.0 L sample
𝑥 = (
84.99 𝑔𝑆𝑜𝑑𝑖𝑢𝑚 𝑁𝑖𝑡𝑟𝑎𝑡𝑒
62 𝑔 𝑁𝑖𝑡𝑟𝑎𝑡𝑒
) ∗ (2 𝐿)∗ (
0.030 𝑔 𝑁𝑖𝑡𝑟𝑎𝑡𝑒
1 𝐿
)
𝑥 = 0.0822
𝑔
𝐿
per every 2 L
5.2.2.5 Determination of 60 mg/L Additional Nitrate
Molar mass of Sodium Nitrate: 84.99 g/mol
Molar mass of Nitrate: 62 g/mol
The sample created was a 2.0 L sample
𝑥 = (
84.99 𝑔𝑆𝑜𝑑𝑖𝑢𝑚 𝑁𝑖𝑡𝑟𝑎𝑡𝑒
62 𝑔 𝑁𝑖𝑡𝑟𝑎𝑡𝑒
) ∗ (2 𝐿)∗ (
0.060 𝑔 𝑁𝑖𝑡𝑟𝑎𝑡𝑒
1 𝐿
)
𝑥 = 1.645
𝑔
𝐿
per every 2 L
5.2.3 Fourth Experiment Calculations
5.2.3.1 Determination of Low Level F/2 Solution (both for Part A and for Part B)
Both Part A and Part B require 0.005 L of Solution to every 38.7 L of water.
The sample required was 1.5 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1.5 𝐿
𝑥 = 194 𝜇𝐿
Low Level of F/2 Solution was determined to only need half of this amount
(0.5) ∗ (194 𝜇𝐿) = 𝑥
𝑥 = 97 𝜇𝐿
5.2.3.2 Determination of Baseline Level F/2 Solution (both for Part A and for Part B)
Part A requires 0.005 L of Solution needed to 38.7 L of water.
The sample created was 1.5 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1 𝐿
𝑥 = 129 𝜇𝐿
5.2.3.3 Determination of High Level F/2 Solution (both for Part A and for Part B)
Both Part A and Part B require 0.005 L of Solution to every 38.7 L of water.
35. The sample created was 1.5 L.
0.005 𝐿
38.7 𝐿
=
𝑥
1.5 𝐿
𝑥 = 194 𝜇𝐿
Low Level of F/2 Solution was determined to need double this amount
(2)∗ (194 𝜇𝐿) = 𝑥
𝑥 = 388 𝜇𝐿
5.2.3.4 Determination of 5 mg/L Additional Phosphate
Molar mass of Potassium Phosphate: 174.18 g/mol
Molar mass of Phosphate: 94.97 g/mol
The sample created was a 2.0 L sample
𝑥 = (
174.18 𝑔 𝑃𝑜𝑡𝑎𝑠𝑠𝑖𝑢𝑚 𝑃ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
94.97 𝑔 𝑃ℎ𝑜𝑠𝑝ℎ𝑎 𝑡𝑒
) ∗ (2 𝐿) ∗ (
0.005 𝑔𝑃ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
1 𝐿
)
𝑥 = 0.0183
𝑔
𝐿
per every 2 L
5.2.3.5 Determination of 10 mg/L Additional Phosphate
Molar mass of Potassium Phosphate: 174.18 g/mol
Molar mass of Phosphate: 94.97 g/mol
The sample created was a 2.0 L sample
𝑥 = (
174.18 𝑔 𝑃𝑜𝑡𝑎𝑠𝑠𝑖𝑢𝑚 𝑃ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
94.97 𝑔 𝑃ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
) ∗ (2 𝐿)∗ (
0.010 𝑔𝑃ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒
1 𝐿
)
𝑥 = 0.0367
𝑔
𝐿
per every 2 L
