Colorful Conclusions: Exploring Paper

Chromatography
Objectives:

• Understand the basic principles behind paper chromatography as a separation technique.

• Gain hands-on experience in conducting paper chromatography experiments, including
spotting, developing chromatograms, and analyzing results.
• Apply chromatography techniques to separate and identify components within a mixture,
utilizing Rf values for comparison with known standards.

Materials:

➢ Chromatography Paper (White Paper Towels can be used as a substitute)

➢ Distilled Water (2 ml)
➢ Ethyl Alcohol (2 ml)
➢ Beakers (3)
➢ Disposable Syringe (1 ml)
➢ Markers (Black, Red, Blue)
➢ Ruler
➢ Pencil Procedures:

1. Cut three individual paper towels into dimensions of 2 centimeters by 15 centimeters each.
Then, measure 4 cm from both ends and draw a line using a pencil and also a midline dividing
the 4 cm line you just drew (as shown in the picture below). This will be the baseline and
solvent front.
*** It’s important to use pencil because lead will not dissolve in water, preventing experimental
error in chemical separation.

2. Using the markers, draw a small dot at the cross section of two lines you have drawn previously.
3. Mix 1 ml of water and 1 ml of ethyl alcohol in the beaker
4. Place the chromatography paper / Paper towel inside the beaker and make sure that the
solvent Does Not reach where the ink is (there should be a gap between ink and solvent). Also,
make sure that the chromatography paper does not touch the bottom of the beaker. Tape
the other end of chromatography paper to a pencil and place it on the beaker

5. Observe what happens as the liquid travels up the paper. Record the changes you see. When the
solvent has reached the pencil line, remove the paper.
TIME SOLVENT REACHED THE SOLVENT FRONT:
Black – 15 sec
Blue – 16 sec
Red – 24 sec
6. Measure how far the solvent traveled before the strip dries.
DISTANCE BETWEEN BASELINE AND SOLVENT FRONT:
Black – 12 cm
Blue – 11 cm
Red – 10 cm
7. Finally, let the strip dry on the desk. With a ruler, measure the distance from the starting point
to the top edge of each color. Record this data in a data table. Calculate a ratio for each color by
dividing the distance the color traveled by the distance the solvent traveled.

 Black – 0.67
 Blue – 0.82
 Red – 1

Color of Ink Distance each Distance the

(Bottom to Top) Color Travelled Solvent
(mm) Travelled (mm)

MARKER 1 Black to pale 8 mm 12 mm

(Black) black

Color of Ink Distance each Distance the

(Bottom to Top) Color Travelled Solvent
(mm) Travelled (mm)
MARKER 2
(Blue) Blue to pale blue 9 mm 11 mm

Color of Ink Distance each Distance the

(Bottom to Top) Color Travelled Solvent
(mm) Travelled (mm)

MARKER 3 Red to pale red 10 mm 10 mm

(Red) (with the hint of
yellow)
LABORATORY OBSERVATION REPORT
ACTIVITY / EXPERIMENT TITLE: Colorful Conclusions: Exploring Paper Chromatography
DATE OF OBSERVATION: MARCH 22, 2024

1. What changes did you observe in the chromatography paper as the liquid traveled up the
paper?
 Nothing appears to happen at first. Then, there is the dot of ink. A thin tideline began to slowly
creep up the paper from the solvent reservoir. The leading edge of the wetted region is known
as the solvent front. The ink dot started to blur as we kept a tight eye on it. This occurs as a
result of the solvent beginning to dissolve the various ink mixture ingredients.

2. How did the ink dots behave as the liquid moved up the paper?
 As the liquid move up the paper, the ink slowly scattered. It’s like the liquid triggers the ink
to spread throughout the wet area of the paper.

3. Did you notice any differences in the behavior of different colors of ink? If so, what were
they?
 Yes, each color of the ink had different time interval before they reached the solvent front.
Also, some colors like the blue and red spread well while the black ink did not scatter very
well. There is also a change of color that occurred in the red ink and the black ink. In the
red ink, we have observes a slight change in color from red to yellow but it wasn’t that
evident. Same as the black ink there was a very light yellow color that appeared after the
black ink has spread.

4. Describe the appearance of the chromatogram once the solvent had reached the solvent
front
 Once the solvent reaches the solvent front on the chromatogram, you would typically
observe separated spots or bands representing different compounds present in the sample.
The appearance can vary based on factors like the type of chromatography used and the
nature of the compounds being separated. These spots or bands may differ in color,
intensity, and position depending on their respective properties and interactions with the
stationary phase.
5. What factors do you think might have influenced the distance traveled by each color of
ink?
 The distance traveled by each color of ink can be influenced by various factors including
the viscosity of the ink, which affects its flow rate, with thicker inks typically traveling
shorter distances compared to thinner ones. Surface tension of both the ink and the paper
can also play a significant role, determining how far the ink spreads before being absorbed
into the paper. Moreover, the chemical composition and concentration of the ink can
impact its behavior on the paper surface. Environmental conditions such as temperature
and humidity can affect the drying time of the ink and its spread on the paper.
 Here are some specific factors that influenced the distance traveled by each color of ink.
1.Solubility
 The ink’s ability to dissolve in the chosen solvent has a significant impact on how
far it can travel. Higher up the paper will be more soluble substances.
2.Paper Affinity
  The ink’s attraction or affinity to the paper can also affect how far it spreads. The
ink might not go as far if it has a strong affinity for the paper.
3.Molecule Size
 Molecules travel a greater distance than larger molecules because larger molecules
move more slowly.
4.Temperature
 The experiment’s operating temperature has the potential to affect its outcomes.
Elevated temperatures have the potential to accelerate capillary action, resulting
in increased ink penetration.
5. Paper Quality
 Kind and quality of paper used can have an impact on the capillary action rate and,
in turn, the ink’s travel time.
6.pH of the Solution
 The ink’s ionization and subsequent interactions with the solvent and the paper
can be influenced by the pH of the solution.
7.Concentration of the Ink
 Ink’s concentration may also be important. In comparison to less concentrated inks,
more concentrated inks might cover less ground.

6. How do you think the ratio of the distance traveled by each color to the distance traveled
by the solvent (Rf value) can help in identifying the components of the ink?
 By measuring the movement of compounds, assigning a distinct value to each compound,
and enabling comparison and analysis of Rf values it can then ascertain the components
present in the ink, after all the Rf value aids in the identification of the ink’s constituent
parts.
7. What conclusions can you draw from your observations about the separation of the ink
components using paper chromatography? What can be the real-life implications of it?
 The water, the paper, and the chemicals that comprise the color are the causes of the
colors’ separation. Pigments are the substances that give a color. Certain pigments go
through the paper more thoroughly before adhering because they cling to water more
readily than others. We observed that As the solvent rises, it dissolves the ink on the paper
and separates the ink into its components.
 In real-life implication, Chromatography is widely used in a variety of fields, including
forensics, petroleum creation, molecular biology, food and beverage production,
pharmaceuticals, clinical trials, environmental and chemical safety, and drug testing.

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