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Paper Chromatography
Introduction
Chromatography is a general technique employed to separate the components of a mixture
based on their relative solubilities in two materials or phases that are set up to move past each
other in essentially opposite direction. One type of this technique is paper chromatography by
which a homogeneous mixture is made to travel through the material of a paper by capillary
action. Capillary action is the ability of a liquid to naturally ascend through a narrow tube or
spaces. A suitable solvent serves as the mobile phase that carries the mixture along the paper,
which is the stationary phase. The solubility of the mixture components between the mobile and
stationary phases influences the degree of the separation of the components. The two phases are
usually chosen so that they have opposite relative polarities (i.e., polar and nonpolar). The
components of the mixture that have similar polarity with the mobile phase travel with it during
the process, while those with similar polarity as the material of the paper are not carried as much.
When the mixture components separate, colored bands or spots form on the paper.

Objectives
 Separate the different pigments of a candy-coating using paper chromatography
 Determine the pigments of a candy coating that have the same nature of polarity as the
mobile and stationary phases used
Materials
different colors of coated candies (e.g., M&M) clear plastic tape
ethyl alcohol pencil
water beaker
filter paper 2 watch glasses
Ruler toothpick
clear plastic

Estimated Time Frame: 40 minutes

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Procedure
1. Cut the filter paper following the dimensions in the illustration. The width of the filter paper
should be about the diameter of the beaker. This is your chromatography paper.
2. Draw a pencil line one centimeter from the bottom of the strip. This line is the “start line”. Do
not use a pen because the ink in pens can interfere in the separation of dyes.
3. Sample Preparation: Take one candy for each color. Place the candies farther apart on a watch
glass. On each candy, place one to two drops of water. Let the samples stand until sample
spotting is to be done.
4. Using a toothpick, make a tiny dot of each color extract along the start line. The dots should be
farther apart. Make sure not to puncture the filter paper with the toothpick.
5. Pour ethyl alcohol into the beaker, just enough to cover the bottom. Then, place another strip
of filter paper (half the size of the chromatography paper) in the beaker so that it sticks to the
side of the beaker. Allow the ethyl alcohol to move up this paper. Cover the beaker with a clean
watch glass.
6. When the entire smaller paper has become wet, carefully place the chromatography paper with
samples in the beaker, making sure that the start line is kept above the ethyl alcohol.
7. Hang the chromatography paper on a pencil and hold it with plastic tape. Cover the beaker
with the plastic wrap.
8. Observe and describe how the ethyl alcohol “climbs” up the paper, dragging the samples with
it. Watch the colored spots go up the strip and possibly separate into individual components.
Write your observations in the activity sheet.
9. Remove the chromatography paper from the beaker when the solvent front reaches the “top
line”
10. Mark where the solvent front is and where the centers of the colored spots are.Use a pencil in
marking the paper.
11. Allow the chromatography to dry. Attach it on a separate sheet of paper and submit to your
teacher.
Observations
How did the appearance of the filter paper change during the progress of the chromatography
procedure?
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Conclusion

Post-Laboratory Questions
1. Is the candy-coating pigment a mixture? Why or why not?

2. How will you use this procedure to identify an unknown type of colored pigment?
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Lighter or Heavier

Objective: Compare the densities of substances

Materials
regular soft drink and diet soft drink in can (same brand)
water
cooking oil
rubbing alcohol
sugar cube
ice cubes
large container
additional materials based on experimental design in part B

Estimated Time Frame: 30 minutes

Procedure
A. 1. Fill the large container with water until nearly full.
2. Place the two cans of soft drink in the water. Record your observations.
Observations
What happened to the cans? Did any can float? If yes, which can? Did any can sink? If yes,
which can?
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B. Design and carry out an experiment to determine the denser substance in each pair of
materials.
1. Water and cooking oil
2. Water and ice
3. Water and sugar cube
4. Rubbing alcohol and ice
5. Cooking oil and ice

Results
Lighter Heavier
Water and cooking oil
Water and ice
Water and sugar cube
Rubbing alcohol and ice
Cooking oil and ice

Conclusion

Post-Laboratory Questions
1. Why is density an intrinsic properly of substance?
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2. A miner discovered some yellow nuggets at the bottom of a shallow stream. The nuggets
weighed 96.5 grams and had a volume of 5 milliliters. Were the nuggets gold or “fool’s gold”
(pyrite)? The density of gold is 19.3 g/mL and that of pyrite is 5 g/mL at 20°C.Show your
calculations to justify your answer.
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Float or Sink
Objective: Determine the density of water and some objects
Materials
top-loading balance
100mL graduated cylinder
water
ruler
stone that will fit inside a 100mL graduated cylinder
ELTS General Chemistry 1 book

Estimated Time Frame: 40 minutes

Procedure
1. Design and carry out a procedure to obtain the density of the book, the stone, and water. Use
only the materials and instruments listed. Write your procedure on the worksheet.
2. Record all your data. Make three trials for each procedure.
Date and Results
A. Obtaining the density of the book
Procedure:
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Data:

B. Obtaining the density of the stone

Procedure:

Data:
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C. Obtaining the density of water
Procedure:

Data:

Conclusion
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Post-Laboratory Question
How can you determine the density of a water-soluble solid like salt or sugar?
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What’s the Product?

Objectives
 Identify the limiting and excess reagents in a reaction
 Determine the percent yield of a reaction
 Determine mass relationships in a reaction
Materials
sodium carbonate (soda ash) two 50 mL beakers
calcium chloride 10 mL graduated cylinder
distilled water weighing balance
2 watch glasses filtration setup
filter paper
Estimated Time Frame :40 minutes
Procedure
1. Obtain two clean and dry watch glasses. Use one watch glass to weigh 3.00 grams of sodium
carbonate, and the other weigh 3.00 grams of calcium chloride.
2. Dissolve sodium carbonate in a beaker containing 10 milliliters of distilled water. Do the same
for calcium chloride but in a separate beaker.
3. Pour the calcium chloride solution to the sodium carbonates solution while stirring.
4. Obtain and record the mass of the filter paper in the data table. Filter the mixture through the
filter paper.
5. Allow the residue to dry. Once dried, weigh the paper with the product. Record your data.
6. Determine the mass of the product formed from the reaction (actual yield).
7. Calculate the theoretical yield of the product based on the initial amount of the limiting
reagent.
8. Calculate the percent yield of the reaction.
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Data and Results
Mass of filter paper
Mass of filter paper with product
Mass of product (actual yield)
Theoretical yield
Percent yield

Conclusion

Post-Laboratory Questions
1. What type of reaction occurred in the activity? Write the balanced equation for the reaction.

2. What is the identity of the solid formed? Justify your answer.

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3. What is the limiting and the excess reagent in the activity? Why do you say so?
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How Fast is the Gas?

Objectives
 Observe Graham’s law of effusion
 Determine the effusion rate of a gas

Materials
rubber tubing (about 6 inches long)
sodium bicarbonate (NaHCO3, baking soda)
acetic acid (CH3COOH, vinegar)
calcium hydroxide solution (Ca (OH)2, limewater)
2 glass tubings (about 3 inches long, with diameter that can fit in a rubber tubing)
ruler
timer
measuring spoon
6-inch test tube
Cork (with hole for glass tubing; should fit into the mouth of the 6-inch test tube)
petri dish
Estimated Time Frame :40 minutes

Procedure
1. Insert the glass tubings at each end of the rubber tubing. Make sure that each glass tubing is
snugly fit into the rubber tubing. Refer to the illustration on page 126D.
2. Measure and record the length from the tip of one glass tubing (end A) to the tip of the other
glass tubing (end B). This will be the distance that the gas will travel.
3. Label one of the ends of the tube as A and the other as B.
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4. Insert end B into the cork. At least one inch of the glass tube should be seen from the bottom
of the cork.
5. Fill the test tube with acetic acid to about one-fourth of its capacity.
6. Fill the petri dish with calcium hydroxide solution to about half its height. Hold end A with the
tip just directly above, but not touching, the surface of the solution.
7. Prepare the timer.
8. Add one teaspoon of sodium bicarbonate to the test tube with acetic. The reaction between
acetic acid and sodium bicarbonate will produce carbon dioxide gas.
9. Immediately cover the test tube with the cork (end B). Start the timer.
10. Stop the timer once you observe a while coloration on the surface of the limewater. This
signifies that carbon dioxide is already present at end A. Record the time it takes for the carbon
dioxide to reach end A.
11. Calculate the rate of diffusion of the carbon dioxide gas as distance over time.
12. Repeat the whole procedure for two more trials. Wash and dry the glass materials in between
trials. Make sure to use the same amount of sodium bicarbonate, and fresh calcium hydroxide,
and acetic acid for each trial.

Observations
Length of the tube(cm):
Trial 1 Trial Trial 3
Time elapsed(sec)
Rate of effusion of
CO2(cm/sec)

Conclusion
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Post-Laboratory Questions
1. Write the balanced chemical equation for the reaction between sodium bicarbonate and acetic
acid to produce carbon dioxide. What is the mole ration between sodium bicarbonate and carbon
dioxide?

2. Will there be a difference in the rate of effusion of carbon dioxide if hydrochloric acid, a
stronger acid, is used instead of acetic acid? Explain your answer.

3. What is the white substance produced as carbon dioxide reaches the calcium hydroxide
solution? Write the balanced chemical equation for the formation of the white substance from the
reaction between carbon dioxide and calcium hydroxide.
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Classifying the Metals

Objectives
 Investigate reactions of ions
 Qualitatively analyze the reactivity of some elements.

Estimated Time Frame :30 minutes

Materials
Sample Solutions: Test Reagents:
sodium nitrate (NaNO3) solution ammonium carbonate[(NH4) CO3] solution
lithium nitrate (LiNO3) solution ammonium hydroxide (NH4OH) solution
calcium nitrate [Ca (NO3)2] solution
barium nitrate [Ba (NO3)2] solution laboratory droppers
copper (II) nitrate [Cu (NO3)2] solution small test tubes
zinc nitrate [Zn (NO3)2] solution test tube rack
nickel (II)nitrate [Ni (NO3)2] solution

Procedure
1. Place five drops of each of the sample solutions in separate test tubes. Label the test tubes with
the corresponding chemical formula of the sample solutions.
2. Place five drops of (NH4) CO3 solution into each of the test tubes. Observe and note which
test tubes form precipitates. Record the color of the precipitates formed.
3. Dispose the chemicals into proper waste bottles. Wash and dry the test tubes.
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4. In a new set of test tubes, place five drops of the sample solutions that formed precipitates in
Step 2.
5. Add three drops of NH4OH solution to these solutions. Observe and record the color of the
precipitate formed.
6. Add 10 more (excess) drops of the NH4OH solution into each test tube.
7. Roll the test tubes in between the palm of your hands to mix the contents. Observe which
among the precipitates will be dissolved.

Observations
Sample +(NH4)2CO3 +NH4OH + excess NH4OH
NaNO3
LiNO3
Ca (NO3)2
Ba (NO3)2
Cu (NO3)2
Zn (NO3)2
Ni (NO3)2

Conclusion

Post-Laboratory Questions
1. Which among the sample solutions did not form a precipitate with (NH4)CO3? What is
common among the cations of these solutions?
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2. Among those that formed a precipitate with NH4OH in Step 5, which sample dissolved in
excess NH4OH in Step 6? What is common among the cations in these samples?

3.What is common among the cations that did not dissolve in Step 6?
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Dissolving Sugar

Objectives
 Explain the solubility of sugar using the concept of polarity
 Observe evidences of molecular polarity

Materials
3 clear plastic cups
3 sugar cubes (same size)
water
cooking oil
70% isopropyl alcohol
white paper
100 mL graduated cylinder

Estimated Time Frame :30 minutes

Procedure
1. Label the three plastic cups: Water, Alcohol, and Oil.
2. Add 30 milliliters of water, alcohol, and cooking oil to the designated cups.
3. Set the three cups on the white sheet of paper.
4. At the same time, add one sugar cube to each liquid.
5. Gently swirl the mixture in each cup for about 30 seconds.
6. Observe how ell each solvent dissolves the sugar and describes the solubility of the sugar.
Write your other observations.
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Observations
Solvent Solubility of Sugar Other Observations
(insoluble, slightly soluble,
very soluble)
Water
70% isopropyl alcohol
Cooking oil

Conclusion

Post-Laboratory Questions
1. Ethyl alcohol is polar, while oil is nonpolar. Based on your observations, what can you say
about the polarity of sugar? Explain your answer.

2. In what other solvents can sugar dissolve? Why?

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Investigating Biomolecules

Objectives
 Observe enzyme action in the hardening of gelatin
 Observe protein denaturation in egg white

Materials
Part A:
gelatin 3 medium-sized bowls egg beater
hard candy blender tablespoons
ripe papaya graduated cylinder
grapes refrigerator
Part B:
5 fresh eggs 70% rubbing alcohol hot plate
3 plastic cups stirring rods pipettes or droppers
Beaker 5% sodium hydroxide
Medium-sized bowl vinegar
Estimated Time Frame: 40 minutes
Procedures
A. Papaya and Gelatin
1. Prepare separate purees of the papaya and grapes. Make sure that the peeling and seeds are
removed before homogenizing the fruit in the blender.
2. Measure about 100 milliliters of each puree and transfer separately into the two bowls.
3. Add two tablespoons of gelatin powder into each bowl. Mix the contents thoroughly using an
egg beater.
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4. In the third bowl, put a piece of the hard candy and add about 50 milliliters of the papaya
puree. Observe and record what happens to the candy.
5. Refrigerate (but do not freeze) the puree-gelatin mixture and check on them the next day. Take
note of the consistency of the mixtures.

Results

Sample Observation(s)

Papaya puree + gelatin

Grape puree + gelatin

Papaya puree + candy

B. Denaturation of Proteins in Egg White

1. Carefully separate the egg white from the yolk of the eggs. Temporarily put the egg white
together in a bowl. You may discard the egg yolks or cook them later.
2. Divide the egg white equally into the three plastic cups and the beaker.
3. Into one plastic cup, place 5 milliliters of vinegar, then mix the contents. Observe and record
what happens to the egg white.
4. In the other plastic cup, place 1 milliliter (about 20 drops) of 5% sodium hydroxide solution.
Mix the contents. Observe and record what happens to the egg white.
5. In the plastic cup, add 5 milliliters of 70% rubbing alcohol, then mix the contents. Observe
and record what happens to the egg white.
6. Place the beaker with egg white on the hot plate and heat at 150°C for not more than 2
minutes. Observe what happens to the egg white during and after heating.
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Results
Step Observation(s)

Adding vinegar

Adding 5% sodium hydroxide

Adding 70% rubbing alcohol

Heating 150°C

Conclusion

Post-Laboratory Questions
1. How is enzyme action evidenced in Part A of the activity?

2. Did your observations in Part B show evidence of protein denaturation? Justify your answer.

3.What structural level(s) of protein are altered during protein denaturation?