Introduction
In this experiment, Pigment Separation in Allium cepa, we will determine among the eluting solutions, such as petroleum ether, water, and acetone, which can seclude the color pigment in red onions.
Aim
To find out which eluting solution can separate the color pigment in red onions.
Theory
1. Rf stands for Retention Factor.
2. The Rf value of the pigment is equal to the distance traveled by the pigment from the origin divided by the distance traveled by the solvent from the origin.
Requirements
1. Red Onions
2. Acetone
4. Distilled Water
5. Chopping Board
6. Distilled Water
7. Plastic Gloves
8. Cotton Yarn
9. Plastic Gloves
10. Plastic Bags
11. Filter Paper
12. Triple Beam Balance
13. Beaker
14. Watch Glass
15. Glass Bottles
16. Graduated Cylinder
17. Pipettes
18. Petri Dishes
19. Wash Bottle
20. Scissor
21. Compass
22. Ruler
23. Notebook
Procedure
Step 1: Prepare the chromatography papers.
Step 2: Prepare red onion extract.
Step 3: Put solvents (petroleum ether, acetone, and water) in Petri dishes.
Step 4: Spot chromatography papers.
Step 5: Observe the movement of pigment in each of the solvents.
Step 6: Record your observations.
Step 7: Perform two experimental batches for each of the three trials, which included petroleum ether, water, and acetone as eluting solutions.
Step 8: Perform four more spots for each of the papers.
Step 9: After drying the papers, note and measure the pigment movement.
Step 10: Now, tabulate, graph, compute and analyze the Rf values of each of the pigments.
Chromatography Papers
Observations
1. From our experiment, we observed that petroleum ether showed no movement of pigments. On the other hand, water and acetone showed the movement of pigments.
2. From the visual analysis of all chromatography papers, we observed that the two pigment colors, purple and yellow, moved in water and acetone.
3. In water dishes, the Rf value of yellow pigment lies between the range of 0.075 and 0.625, with a mean of 0.174.
4. In water dishes, the Rf value of purple pigment lies between the range of 0.875 and 0.975, with a mean of 0.932.
5. Only yellow color pigment moved in three of the six acetone dishes. There, it had an Rf value that ranged between 0.800 and 0.975. It acquired a mean of 0.454.
Result
1. The pigment got separated into two pigment colors. This includes yellow (quercetin) and purple (anthocyanin).
2. From the movement of pigments in all six trials in the eluting water solution, we concluded that the pigments are polar in nature.
3. Acetone eluting solution moved the yellow pigment in three among six of the trial; this indicated the weakly polar nature of pigments.
Precaution
1. Handle the eluting solutions with care.
2. Record your observation carefully.
Conclusion
In this experiment, Pigment Separation in Allium cepa, we determined the movements of colored pigment in three of the eluting solutions.
VIVA Questions With Answers
Q.1 What was the aim of your experiment?
ANS. We aimed to determine among the eluting solutions, such as petroleum ether, water, and acetone, which can separate the color pigment in red onions.
Q.2 In which eluting solution pigments showed polar nature?
ANS. In the eluting water solution, pigments showed polar nature.
Q.3 Which eluting solution showed the weak polar nature of pigments?
ANS. Acetone showed the weak polar nature of pigments.
Q.4 Which eluting solution showed no motion of pigments?
ANS. Petroleum ether elution solution showed no motion of pigments.
Saquib Siddiqui is a Mechanical Engineer and a content creator. He has special interest in science experiments and teaches the same to school and college students. He is also a technical lead at RUDE LABS PVT. LTD.
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