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Anticoagulants
March 26, 2021
Therese Nguyen
Mrs. Zander
8th Science

Abstract

The purpose of this experiment is to compare how blood coagulates with an anticoagulant through

solutions of sodium alginate forming into semisolid balls using 3 different concentrations of sodium citrate. The

question asked was, “How can an anticoagulant affect blood clotting?” 240 milliliters of water was added into 3

bowls, followed by 1.3 grams of calcium chloride. Similar to blood, the sodium alginate needs a source of calcium

to coagulate or create semisolid balls. Blood can be a hazardous substance and is hard to obtain without

anticoagulants already in it. Using sodium alginate is a safer and more achievable alternative to using actual blood.

The control of this experiment was the “no sodium citrate” solution. The sodium citrate acted as an anticoagulant.

Anticoagulants disrupt the blood coagulation process. The control is compared to the 1% sodium citrate solution

and the 1.5% sodium citrate solution. There were 5 balls per solution, and the diameters and heights were measured

via graph paper. The results were found by averaging. The hypothesis that was tested is “If the source of calcium

chloride solution is used on the sodium alginate solution, semisolid balls will then form, compared to when sodium

citrate is added, the semisolid balls will not form.” It was somewhat supported in this experiment. Semisolid balls

did form in all the solutions, but as more sodium citrate was added, the ball’s height decreased while the widths

increased. They became less like spheres and it took longer for the balls to congulate.

Introduction

Let it be a cut, a scratch, or a wound, everyone has experienced external bleeding at least

once in their lifetime. Blood is an essential liquid that transports nutrients and oxygen throughout

the body using blood vessels. It also transports wastes products away from those same cells.

Blood is made up of four main units; red blood cells, platelets, plasma, and white blood cells.
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Bleeding, also known as hemorrhage, occurs in the body when a blood vessel or blood

vessel wall has been damaged or breaks. Though bleeding may seem bad, that is not always the

case. Blood can bring out important components to help clean wounds. On the contrary, bleeding

too much is dangerous and can cause your body to go into shock. Your heart would be unable to

pump the sufficient amount of blood throughout your body if you lose enough of it. To prevent

this and to allow the wound to heal, the bleeding must stop through a process called hemostasis.

An important part of hemostasis is the blood coagulating, also known as blood clotting.

There are 4 main components needed in hemostasis, which can be normally found in the

blood; platelets, clotting factors, fibrin, and specific cell types. When you bleed, platelets, small

fragments of cells that stick to the opening in the blood vessel, rush to the opening of the wound.

When enough of them stick together, it forms a temporary plug for the wound. The drawback to

this is that the clot is nowhere near as stable as it needs to be. To reinforce the platelets, clotting

factors from the blood join the platelets in the clot. To reinforce the clot even further, the clotting

factors create fibrin. Fibrin is a string-like protein that holds the other factors in place and forms

a mesh throughout the clot. Red and white blood cells also contribute in forming the clot. Once

the wound is healed, the clot is naturally dissolved.

When the body cannot perform this correctly, it is known as a blood disorder, and can be

a serious health hazard. One example is hemophilia, a disorder where the body has difficulty

clotting blood. This can cause the person to bleed excessively, internal and external. To treat a

person with hemophilia, they are often injected with the clotting factors that they are missing.

Though blood clotting is very important, there are some instances where the process must

be prevented. Unwanted blood clots can clog blood vessels, which stops blood from flowing to

major organs. This can cause a range of health difficulties, such as a heart attack or a stroke. To
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help avoid this, patients with this problem are prescribed anticoagulants. Anticoagulants work by

disrupting the blood clotting process. An example of a commonly used anticoagulant would be

trisodium citrate. During blood clotting, platelets increase the amount of calcium around the

wound, which affects the different proteins and how they interact with the clotting factors. In a

chemical reaction, the trisodium citrate grabs the calcium. It does this by breaking apart into

sodium and citrate, and then having the citrate bind to the calcium, thereby forming calcium

citrate. This means that there no longer is an increased amount of calcium present, successfully

ending the coagulation process.

This experiment will be investigating the question, ‘How can an anticoagulant affect

blood clotting?’ It will use sodium citrate as an alternative to trisodium citrate, since it is easier

to obtain and is safer to use. Instead of using real blood, which is dangerous and harder to

purchase without anticoagulants, a solution of sodium alginate and calcium chloride will be used.

Sodium alginate is a hydrocolloid (moisture-retentive dressings, which contain gel-forming

agents) sourced from brown seaweed. Similar to blood, sodium alginate requires calcium to

congulate. Adding it into calcium chloride will cause it to form a semi solid ball. This happens

through a chemical reaction that rearranges the alginate molecules into bonding with the calcium

ones.

2NaC6H7O6 + CaCl2 → 2NaCl + C12H14CaO12

The equation above shows the chemical reaction between sodium alginate and calcium

chloride, creating calcium alginate, a gelatinous substance. The hypothesis to this experiment is

that if the source of calcium (Calcium chloride solution) is used on the ‘blood’ (sodium alginate

solution), semisolid balls will form, compared to when sodium citrate is added, the semisolid
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balls will not form. The goal is to compare the formulation of these balls, with and without

sodium citrate.

This experiment adds to the field of hematology as more of a subject of testing. Using the

results provided, more efficient medication can be derived, along with improved studies that

could potentially lead to bigger or smaller clots.

Materials

● Cold water
● Measuring cup (mL)
● Wax paper
● Food coloring
● Plastic wrap
● Bowls (3)
● Sticky notes
● Spoons
● Timer
● Scale
● Graph
● Camera
● Notebook
● 1mm graph paper
● Blender
● 60 cc plastic syringe
● Sodium alginate (2g)
● Calcium chloride (4g)
● Sodium citrate (6g)

Procedure
1. The balls will be quantified by placing them onto 1mm graph paper and determining the
dimensions (height and diameter). The first step to this experiment is preparing the items
used to measure these semi solid balls. Since graph paper may have some extra edges that
can cause some inaccuracies in the quantifying process, take the 1mm graph paper and
trim the edges, leaving the first lines of the grid to be at the edge of the paper.
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2. In the lab notebook, create this data taking table.

3. Create the sodium alginate solution by pouring 120 mL of water into a blender. Weigh 2
grams of sodium alginate (use wax paper on the scale because it will be harder for the
sodium alginate to stick to), and then add it with the water. Add 5 drops of food coloring,
and to top it all off, add another 120 mL of water.
4. Once the water, sodium alginate, and food coloring have been prepared, blend so the
solution is fluid and homogeneous.
5. The next step is creating the calcium chloride solutions with different concentrations of
sodium citrate. Set up 3 bowls and label them all with, “No sodium citrate” (the control),
“1% sodium citrate”, and “1.5% sodium citrate”. Add 240 mL of water into each of them.
Then, add 1.3 g of calcium chloride to each bowl. Measure it out the same way the
sodium alginate was measured out in step 3.
6. To the bowl labeled “1% sodium citrate”, add 2.4g of sodium citrate. To the bowl labeled
“1.5% sodium citrate”, add 3.6g of sodium citrate. Stir each bowl using a new, clean
spoon to avoid cross contamination until the solutions are dissolved.
7. Before doing the experiment, test if you can make sodium alginate balls using your
different calcium chloride solutions to ensure that it has been done correctly.
8. Prepare the timer.
9. Using the syringe, suck up a small amount of sodium alginate solution. If there is a layer
on top, dip the syringe below the layer to only suck up the liquid part. Wipe off any extra
foam on the syringe. Practice releasing the syringe to only form 1 drop at a time.
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10. Once drops can be successfully made, drop a single drop into the bowl containing the
solution of calcium chloride without sodium citrate. The tip of the syringe should be
around 8-13 cm above the surface of the solution. Set the timer for 60 seconds and let the
drop sit in the solution. After the timer is up, try to scoop the ball out of the solution as
perfectly as possible.
11. Place plastic wrap over the graph paper in order to measure the diameter of the ball.
Based on how many lines the ball spans, calculate the ball's diameter in mm. The
diameter of the ball can be measured by carefully moving the plastic wrap so that one
edge of the ball is lined up on a line.
12. If the ball is not a sphere, measure it’s longest diameter and it’s shortest diameter and
record both diameters in the data table. If, however, it is a sphere, record the same
number for both diameters. If there is excess liquid making the ball hard to measure, dab
it with a paper towel. If the ball gets damaged in the process, make a new one.
13. Measure the height of the ball by placing the graph paper prepared in step 1 behind the
ball. Make sure you are eye level with the ball while measuring it. Record the height into
the data table.
14. Repeat steps 10-14 4 more times so that you have measured 5 balls using the solution of
calcium chloride without sodium citrate.
15. Repeat steps 10-14 until 5 balls can be measured, this time using the 1% sodium citrate
solution.
16. Repeat steps 10-14 until 5 balls can be measured, this time using the 1.5% sodium citrate
solution.
17. (Optional) Take pictures of the results.

Data

The table shown shows the diameters of the balls, their heights, and the average of both

the heights and diameters. During this experiment, everything was kept as consistent and the

measurements were estimated as accurately as possible. If the ball was a sphere, the “longest”

and “shortest” diameters would be the same. If it was not, then they would be different.
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The blue graph below is a bar graph of the averaged diameters for the 5 balls in each

solution in millimeters.

The green graph below is a bar graph of the averaged heights for the 5 balls in each solution in

millimeters.
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Results

The data was collected and averaged to get the results.

Average Formula: Total Sum of All Numbers/Number of Items in the Set

Average Diameter

No Sodium Citrate:

Ball 1: 4+4, divided by 2= 4 mm

Ball 2: 4+4, divided by 2 = 4 mm

Ball 3: 5+5/2, divided by 2 = 5 mm

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Ball 4: 5+4.5, divided by 2 = 4.75 mm

Ball 5: 5.5 + 5.5, divided by 2 = 5.5 mm

Total Average: 4+4+5+4.75+5.5, divided by 5 = 4.65 mm

1% Sodium Citrate:

Ball 1: 5+5 = 5, divided by 2 = 5 mm

Ball 2: 6+5 = 5.5, divided by 2 = 5.5 mm

Ball 3: 7 + 6.5, divided by 2 6.75 mm

Ball 4: 6 + 5, divided by 2 =5.5 mm

Ball 5: 7 + 7, divided by 2 = 7 mm

Total Average: 5 + 5.5 + 6 .75 + 5.5 + 7, divided by 5 = 5.95 mm

1.5% Sodium Citrate:

Ball 1: 8.5 + 7.5, divided by 2 = 8 mm

Ball 2: 9 + 6.5, divided by 2 = 7.75 mm

Ball 3: 6 + 5.5 , divided by 2 = 5.75 mm

Ball 4: 7 + 5.5, divided by 2 = 6.25 mm

Ball 5: 9 + 8.5 = 8.75, divided by 2 = 8.75 mm

Total Average: 8 + 7.75 + 5.75 + 6.25 + 8.75 = 7.3mm

Average Height

No Sodium Citrate: 3 + 4 + 4 + 5+ 4.25 = 4.05 mm

1 % Sodium Citrate: 4 + 4 + 3.5 + 3 +3 = 3.5 mm

1.5% Sodium Citrate: 3 + 3 + 1.5 + 1.5 + 1.5 = 2.1 mm

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Average Longest Diameter:

No Sodium Citrate: 4 + 4 + 5 + 5 + 5.5, divided by 5 = 4.7 mm

1% Sodium Citrate: 5 + 6 + 7 + 6 + 7, divided by 5 = 5.2 mm

1.5% Sodium Citrate: 8.5 + 9 + 6 + 7 + 9, divided by 5 = 7.9 mm

Average Shortest Diameter:

No Sodium Citrate: 4 + 4 + 5 + 4.5 + 5.5 = 4.6 mm

1% Sodium Citrate: 5 + 5 + 6.5 +5 + 7 = 5.7 mm

1.5% Sodium Citrate: 7.5 + 6.5 + 5.5 + 5.5 + 8.5 = 6.7 mm

Through these results, the question was successfully answered. Anticoagulants do affect blood

clotting by slowing it down. The effect can be seen by looking at the diameters and heights,

along with looking at the semi balls up close.

Discussion

Congulation is an important process to understand in the medical field. It is essential to

breaking down and treating blood disorders. Disrupting blood clotting can cause these disorders

since it cannot coagulate properly.

As more sodium citrate was added, the balls got less firm and their heights decreased.

Their diameters, however, increased. They slowly became less sphere shaped, and more like oval

blobs. In the no sodium citrate bowl, sturdy semisolid balls formed. The average diameter was

4.65 mm, and the average height was 4.05mm. In the 1% sodium citrate bowl, the balls became

less firm and more liquid. The average diameter was 5.95 mm, and the average height was 4.05

mm. In the 1.5% bowl, the balls took longer to coagulate and were very jelly like. The average

diameter was 7.3 mm, and the average height was 2.1 mm. This is a result of sodium citrate
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interfering with the congratulation process by binding with the calcium, as stated earlier in the

introduction. It makes the balls less firm and not hold up as well, also known as evidence that

the congratulation process was disrupted. The averages of both the heights and diameters shows

the results of this experiment, and how each solution differs from one another. This proves the

hypothesis, “When the source of calcium calcium chloride solution is used on the sodium

alginate solution, semisolid balls will form, compared to when sodium citrate is added, the

semisolid balls will not form.” only somewhat supported. Semisolid balls did form in the “No

Sodium Citrate” bowl and in the bowls with sodium citrate. It was just less apparent and less

structured for the bowls with sodium citrate. This end product wasn’t fully expected, and it was

thought that the sodium citrate would completely prevent the semisolid balls from forming. The

error in the hypothesis was that the sodium citrate didn't completely stop the congratulation

process, it simply slowed it down. The more sodium citrate added the easier the balls could be

broken. Despite the unexpected outcome, this project was still a success. There was a significant

difference in the 3 solutions, especially the one without sodium citrate. The shortest and longest

diameters were also averaged, as seen in “data”.

The experiment performed could’ve been performed more accurately if the increments

used were more specific. Unfortunately, graph paper that provided decimal millimeter

increments would be hard to obtain. Using more exact tools, such as a blender with mL

measurements, would’ve helped make the data taking more precise.

A hematologist could explore this data and research it further. They could develop this

procedure more in depth and have research done on this experiment to increase the efficiency of

blood clotting and anticoagulants. Hematologists are people who diagnose people with blood

disorders and prescribe the appropriate medication for these things. They could use the
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knowledge gained from this experiment to try to improve the diagnosing and treatment of these

disorders.

Conclusion

In this experiment, the question answered was “‘How can an anticoagulant affect blood

clotting?” Anticoagulants can slow down the coagulation process, and can prevent the blood

from clotting. The overall objective was to show how the 3 solutions differed from each other by

comparing the semisolid balls formed in each. In the no sodium citrate bowl, firm balls of

calcium alginate formed. The average diameter was 4.65 mm, and the average height was 4.05

mm. In the 1.0% bowl of sodium citrate, the semisolid balls congulated slower and were less

firm. They still held up, but were much easier to break. The average diameter was 5.95 mm, and

the average height was 3.5 mm. Lastly, in the 1.5% solution, the semisolid balls did not form as

well and were barely congulated. They were very jelly and liquidy. The average diameter was 7.3

mm, and the average height was 2.1 mm.

The hypothesis, “If the source of calcium calcium chloride solution is used on the

sodium alginate solution, semisolid balls will then form, compared to when sodium citrate is

added, the semisolid balls will not form.” was proven half corrected, as stated earlier in the

discussion. Semisolid balls did form in all solutions, but as more sodium citrate was added, the

process became some just slower. Investigating blood coagulation using these solutions can be

beneficial to people who involve themselves with blood and human biology.

This experiment could be taken a step further with the usage of actual blood and

anticongelantes. It could even begin to ask the question, “How can one advance anticoagulants?”

It taught what anticoagulants could do to blood.

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Personal Reflection

This science fair project was very enjoyable to do. I personally wish I would’ve done it
sooner to avoid rush, but it all worked out in the end. I chose this project to get a small glimpse
of what to study in case I would like to enter the medical field, and to learn more about human
biology, specifically blood. It was a valuable learning experience for me because not only did I
improve my ability to plan a manageable schedule, but I was able to do a better job researching
information online. Being precise was a very important step, and I had checked my data multiple
times to make sure there was as little error as possible. Overall, I am very proud of my work.

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