General Chemistry

Laboratory Activity 1

Paper Chromatography

Group No: 1 Date: 10/20/21

Name: Mendoza, Patrice Berlen G. Grade and Section: STEM 12 A

(NOTE: Submit your paper chromatogram after experimenting!)

Chromatography is a method of separating mixtures by means of their affinity. The affinity can
be based on the solubility or polarity of the components towards the solvent. All forms of
chromatography have two components or phases: the mobile phase and stationary phase. The mobile
phase is the phase that moves in a definite direction. The stationary phase is a substance or material
fixed in place in the chromatography procedure. This is where the mobile phase moves.

Paper chromatography is a form of chromatography that uses paper as the stationary phase. A
sample of mixture is placed in the paper and allowed the eluent (the solvent) to move via capillary
action (the ability movement of liquid to move across the narrow spaces without any aid). The eluent
separates the samples into components or pigments. The retention factor or Rf value is computed to
determine whether the component has an affinity towards the solvent or the mobile phase. The Rf of
zero means the component is immobile and has no affinity. The Rf of 1 or less (but not equal to zero)
means that the component has an affinity to the mobile phase and travels with the solvent front. The
longer the distance covered by a component, the greater the Rf value, the higher its affinity towards the
solvent.

Objectives:

 To perform the paper chromatography activity

 To measure the Rf values of different pigments
 To describe the affinity of the component pigments/colors based from the chromatogram.

Materials:

1 pc. 5” x 2” bond paper Distilled water

1 pc. 250-ml beaker or clear glass or glass jar Clear tape

1 pc. 50-ml graduated cylinder or syringe Sharpened pencil

1 pc. petri dish cover or saucer Ruler

1 pc. barbecue stick 1 Sign pen of any color (black or red, MyGel
brand is preferred)
70% ethyl alcohol

Procedure:

A. Developing Chamber making

1. In a beaker, pour 20 ml (or 2/3 of the total volume) ethyl alcohol solution and 10 mL water (1/3
of the total volume, or any amount enough to submerge the small portion of the paper) onto
the glass jar. Use the syringe.
2. Swirl the mixture, but DO NOT SPLATTER!
3. Cover the mouth of the jar with the saucer. Allow the vapors to fill in the remaining space of the
beaker.
B. Paper Chromatogram making
 1. GetS the 5” x 1 ½ ” bond paper. Careful not to touch the middle of the paper, as it may affect the
separation process.
2. In one side of the paper, draw a straight line (about ¼ inch from the edge) using a ruler. Place
one X on the middle of the straight line. See the image below. This is now your chromatogram.

Figure 1. Paper Chromatogram

3. Attach the other side of the chromatogram to a barbecue stick. Secure it with a clear tape.
C. Paper Chromatography
1. Create a blot of sample by placing the tip of the sign pen at the middle of the X-mark, gently. Do
the blotting twice. DO NOT OVER BLOT!
2. Allow the ink sample in the paper chromatogram to dry.
3. Remove the petri dish. Slowly dip the lower side of the paper chromatogram into the solution at
the same time. The samples along the straight line SHOULD BE ABOVE THE LEVEL OF THE
SOLUTION. Cover back the mouth of the beaker using the petri dish, as seen on the figure
below.

Figure 2. Paper Chromatogram inside of the developing chamber

4. Leave the developing chamber for 45 minutes or more. Notice that the inks begin to separate
into two or more colors as the solvent or eluent moves up.
5. As soon as the color separates completely, stop the eluent from running upwards by removing
the cover and gently removing the chromatogram. Allow it to dry.
6. Draw the line of the solvent front of the chromatogram using the ruler and pencil. Measure the
distance covered by the solvent from the straight line to the solvent front in centimeter.
Distance covered by the solvent front: 3 cm
7. Identify the colors found in the chromatogram. Encircle or elongate the first color nearest to the
solvent front (Color A). Locate the center of the elongated color and place a dot on it. Measure
the distance covered by each color from the straight (starting) line to the dot of each color in
centimeter. Do the same for the other pigments found.
i. Color of pigment A: ____Blue____ Distance covered by Color A: ___3.2 cm__
ii. Color of pigment B: ____Pink____ Distance covered by Color B: ___2.5 cm___
 iii. Color of pigment C: ____________ Distance covered by Color C: ___________
iv. Color of pigment D: ____________ Distance covered by Color D: ____________
(Note: Fill up only iii and iv if there are more than two colors appeared from the chromatogram.)
8. Compute the retention factor or Rf value for every color component using the equation below.
Show your solution inside of the box.
distance covered by the component
Rf value=
distance covered by the solvent ∨eluent

Solution for Color A Solution for Color B

3 .2 cm 2.5 cm
Rf value= Rf value=
3 cm 3 cm

3.2 cm 2.5 cm
¿ ¿
3 cm 3 cm

Rf value=0. 9 375∨0.94 Rf value=0.83

Solution for Color C Solution for Color D

Questions:

1. In your activity, which is the mobile phase? Stationary phase?

- In our activity, the solvent is the mobile phase, which is the combination of
20 mL ethyl alcohol and 10 mL water as it tends the movement of the
pigment through the paper. While the paper is the stationary phase, in which
the mobile phase moves through and the pigments scatters.

2. What are the component colors/pigments in Color A? Color B? Color C? Color D?

- With the use of a blue pen, colors (A) Blue and (B) Pink can be observed.

3. Which component colors/pigments has the longest distance covered?

- The pigment that has the longest distance covered is Color A (Blue).

4. Which component has the greatest Rf value? Lowest Rf Value?

- The component that has the greatest Rf value id Color A (Blue) with a total of
0.9375 or 0.94. While the component that has the lowest Rf value is Color B
(Pink) with a total of 0.83.

5. If your ink sample did not move across the stationary phase, how can you describe the
affinity of such ink to your mobile phase?
- If the ink sample did not move across the stationary phase it is an indication
that the ink has a weak affinity or by chance – no affinity to the mobile
phase. Basically, the ink's components won't be attracted to the eluent,
resulting for the stationary phase to be adhered instead thus the ink spot
wouldn't budge in its place.
IMPORTANT NOTE:
1. Attach your paper chromatogram inside of the box. One of the members should bear the original
paper chromatogram, while the rest will paste the photo of their original chromatogram.

2. Attach at least one photo of your actual laboratory performance inside of the box.