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Vishal Parmar Ms. Frye Science October 21st, 2013 Research Plan Question: How will adding different amounts of anticoagulants affect the coagulation process to create varying blood disorders? Hypothesis: If varying amounts of anticoagulants are added into the coagulation process (process of turning liquid to semi-solid/solid), then the coagulation process (blood clotting) will be disrupted because anticoagulants turn semi-solids into liquids. Variables: Independent: The amount of sodium citrate being added to the calcium alginate solution Dependent: What forms when different amounts of sodium citrate is added to the calcium alginate solution

Materials: Graph Paper (one sheet) 8-10cm wax paper (two sheets) 1 cup of cold tap water Digital Scale (measure to .1 gram increments) 2 grams of Sodium Alginate Clean spoon (at least 7 different spoons) Food Coloring (5 drops) Blender (big or small)

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3 bowls

Sticky Notes

1 cup of room temperature water

3.9 grams of Calcium Chloride Plastic Wrap

5 grams of Sodium Citrate Log Book

Syringe

Stopwatch

Pencil/Pen

Procedures I. Measure Ball using graph paper (cut off ends where there are not full 2mm spacing apart assuming you graph paper is 2 mm spaced) (you are going to measure the height and diameter of the balls)

II. Make a graph like the one below Diameter (mm) Solutions Ball Longest Diameter No Sodium Citrate 1 2 3 Average 1% Sodium 1 2 3 Shortest Diameter Average Height (mm) Observations

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Citrate 1.5% Sodium Citrate

Average 1 2 3 Average

III. Make 9 total Coagulant Balls (See Steps below to do so) - 3 with no Sodium Citrate -3 with 1% Sodium Citrate -3 with 1.5 Sodium Citrate -We are seeing if any balls form NOTE: The reason we chose the percentages we chose is because we want to observe how even the smallest increase in Anti-Coagulants can affect Coagulation.

IV. Now make the Sodium Alginate solution. 1. In blender, add cup (120 mL) of cold tap water. 2. Weigh out 2 grams of sodium alginate and add that to the water in the cup. A. To weigh out the sodium alginate and other chemicals used in this science project, cut a small piece of wax paper (around 8 cm10 cm on each side), place the wax paper on the scale, make the scale read 0 grams, and then weigh out the chemical on the wax paper. B. Use a clean spoon to scoop the chemicals out of their containers. Note: You should use wax paper because it is harder for chemicals to stick to than normal paper. C. Tip: If the scale you are using does not have a feature to zero it out, you will need

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to first weigh the piece of wax paper so that you can subtract this weight from the total when you weigh the chemicals on it. 3. Add five drops of food coloring to the blender cup. 4. Add another cup (120 mL) of cold tap water to the blender. (Adding the rest of the water now should help mix the sodium alginate and food coloring a little) 5. Tip: To help mix solution, blend contents 2-3 times for 5-10 seconds each. 6. The solution should homogeneous (All parts of the solution are mixed and it looks the same)

V. Now make the Calcium Chloride Solutions. There will be 3 sets, each with different concentrations (amounts/ratios) of sodium citrate i.e. 0%, 1%, 1.5% 1. Set out 3 bowls and label them with a sticky note or piece of paper: 0% Sodium Citrate,1% Sodium Citrate, 1.5% Sodium Citrate 2. Add 1 cup of room temperature water (240 mL) to each bowl. 3. Add 1.3 grams of Calcium Chloride to each bowl A. To measure, be sure to even out the scale so it reads 0 grams, then take a fresh sheet of wax paper (so the Sodium Alginate doesnt interfere with the Calcium Chloride) to weigh it. 4. Add 2.4 grams of Sodium Citrate to bowl labeled, 1% Sodium Citrate A. Measure Sodium citrate as done in step V.3.A. B. You weigh out 2.4 grams of Sodium Citrate because 240 (amount of water in C. each bowl) multiplied by .01 (1%) equals 2.4

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5. Add 3.6 grams of Sodium Citrate to bowl labeled, 1.5% Sodium Citrate A. Measure Sodium citrate as done in step V.3.A. B. You weigh out 3.6 grams of Sodium Citrate because 240 (amount of water in each bowl) multiplied by .015 (1.5%) equals 3.6 6. Stir the components of the solution in each bowl. *Note: Use a different and clean spoon each time to avoid contaminating the other solutions.

VI. Test to see if it is possible to make Calcium Alginate balls. Make a total of 3 balls for each solution with different concentrations of Sodium Citrate (9 balls total). 1. Have graph paper use in step I, and a stopwatch available and ready for use. 2.Use syringe to suck up a small amount of the Sodium Alginate solution (Be sure to only suck up liquid. If there is foam above the Sodium Alginate Solution, suck up liquid below the foam) 3. Release one drop of Sodium Alginate from syringe 3-5 inches away from bowl labeled, 0% Sodium Citrate A. Using the stopwatch, let the Sodium Alginate sit in the Calcium Chloride solution for 60 seconds to allow for the Sodium Alginate to dissolve. B. After 60 seconds, scoop out about a small spoonful (using a clean spoon) of the Calcium Alginate Solution (try to scoop as little as possible while still making a ball) i. When scooping out the solution, if a ball or if anything doesnt form, record 0 for the diameter and height in your notebook a. The measurements for the chemicals were chosen so a semi-solid

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ball DOES form. If it doesnt form, recheck measurements C. Measure the diameter of the Calcium Alginate Ball by placing it on plastic wrap and then placing it on the graph paper. i. Line the ball with the edges of the lines on the graph paper. ii. If the graph paper has lines that are 2 mm apart, then multiply 2 by the number of lines the ball spans. a. If the ball is not a sphere, then find out its longest and shortest diameter and record them both. If the Ball is a sphere, then find out the diameter and record the same number for its longest and shortest diameter. b. If there is any liquid around the ball, take a dry paper towel and dab the liquid. If you damage the ball while doing this, make a new ball and do not use this one for the project. D. Measure the height of the Calcium Alginate Balls by placing graph paper behind the ball. i. Lower your eye to where the ball and graph paper (lower eye to the surface you are using) ii. Record height as in the same manner you recorded the diameter (reference to step VI.C.ii) E. Repeat steps VI.3.B - VI.3.D.ii two more times so you have measured a total of 3 balls with 0% Sodium Citrate. F. Repeat steps VI.2 - VI.3.D.ii a total of 3 times (so that you make a total of 3 balls) except use the bowl labeled 1% Sodium Citrate. Be sure to record all

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observations. G. Repeat steps VI.2 - VI.3.D.ii a total of 3 times (so you make a total of 3 balls) except use the bowl labeled 1.5% Sodium Citrate. Be sure to record all observations.

VII. Analyze the balls 1. Record how the ball looks and feels 2. Looking at the graph made, find the average diameter for each ball by dividing the sum of the largest and shortest diameter by two for each ball. 3. Average the heights of all in each of the solutions (have a separate averages for each solution with different concentrations of Sodium Citrate). To find out how to average, reference to step VII.2. 4. Average the diameters for each ball of each solution (ex: Average the three diameters from 0% Sodium Citrate. Do the same with 1% Sodium Citrate and 1.5% Sodium Citrate. You should end up with 3 different diameters. 5. Now create a double bar graph to visualize all the information. A. Put the three different solutions under the X-axis B. Now name the Y-axis Average Heights and Diameters (in millimeters). C. Make sure to color code the bars if they are signifying the height or the diameter. 6. Now analyze the data by considering the following questions: A. How did the balls change with more Sodium Citrate being added? B. Do the results mean that the Sodium Citrate (Anti-Coagulant) disrupted the

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Coagulation process? i. Did the diameter and height of the balls both increase, decrease, or did they have opposite effects? What can you infer from how the balls general shape changes? C. What does the results tell you about how Anticoagulants work and affect Coagulation? D. Based on results, how is Coagulation important? How can adding Anticoagulants affect the Coagulation process to cause varying blood disorders (like hemophilia)? E. Are anticoagulants always harmful? If not, then in what ways can they be beneficial? F. If there were 2% Sodium Citrate in the Calcium Alginate solution what would you expect to form? Would a semi-solid ball form?

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Works Cited
"American Society of Hematology." Bleeding Disorders. American Society of Hematology, n.d. Web. 20 Oct. 2013. <http://www.hematology.org/Patients/Blood-Disorders/5219.aspx>.Bingham, Caroline, et al. Pump That Blood! Human Body. New York City: DK, 2003. 19-21. Print.

Hoare, Ben, et al. "Blood." The Kingfisher A-Z Encyclopedia. New York City: Kingfisher Publications, 2002. 44+. Print.
"How Does Blood Clot?" Circulatory System - How Does a Blood Clot Form? Neok12, n.d. Web. 20 Oct.2013.<http://www.neok12.com/php/watch.php?v=zX020b007271626a7e001a45&t=Circula tory- System>. Parker, Steve. Blood Circulation. Human Body. New York City: DK, 1993. 30-34. Print. Rowland, Teisha, and Andrew Bonham. "Blood Clotting to the Rescue: How to Stop Too Much Blood from Flowing." Blood Clotting to the Rescue: How to Stop Too Much Blood from Flowing. Science Buddies, n.d. Web. 20 Oct. 2013. <http://www.sciencebuddies.org/science-fairprojects/project_ideas/HumBio_p037.shtml>. "What Causes Excessive Blood Clotting?" What Causes Excessive Blood Clotting? NIH, 30 May 2012. Web. 20 Oct. 2013. <http://www.nhlbi.nih.gov/health/health-topics/topics/ebc/causes.html>.