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2021 Expt 11 Pre Lab - Chromatography

This document provides instructions for a chromatography lab module. The learning outcomes are to explain chromatography technique and calculate Rf values. Students will use paper chromatography to separate pigments from plant extracts using different solvents. By measuring the distance traveled by each pigment spot from the origin and dividing by the distance traveled by the solvent front, students can calculate Rf values and determine the relative molecular weights of the pigments. Proper technique such as using a pencil to mark spots and avoiding immersing the origin in solvent is important to get accurate results.

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NUR NAJWA BINTI MOHD RAFIE Moe
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1K views4 pages

2021 Expt 11 Pre Lab - Chromatography

This document provides instructions for a chromatography lab module. The learning outcomes are to explain chromatography technique and calculate Rf values. Students will use paper chromatography to separate pigments from plant extracts using different solvents. By measuring the distance traveled by each pigment spot from the origin and dividing by the distance traveled by the solvent front, students can calculate Rf values and determine the relative molecular weights of the pigments. Proper technique such as using a pencil to mark spots and avoiding immersing the origin in solvent is important to get accurate results.

Uploaded by

NUR NAJWA BINTI MOHD RAFIE Moe
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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SB025 Pre-Lab Module

EXPERIMENT 11: CHROMATOGRAPHY

Course Learning Outcome:


Solve basic problems related to transport system processes, mechanisms for adaptations in
living things, ecological and environmental issues in biology.
(C4, PLO 4, CTPS3, MQF LOD 6)

Learning Outcomes:
At the end of this lesson, students should be able to:
i. Explain the concept of chromatography technique.
ii. Calculate the Rf values

Student Learning Time:


Face-to-face Non face-to-face
1 hour 1 hour

Direction: Read over the lab manual and then answer the following questions.

Check this out:

Each object in our surrounding has their own colour. Let’s have a
look at this green marker. Have you ever wonder why it is green?

Let’s spare a few minutes to watch this short video about Dr.
Brian Mahon asks a graduate student Venecia Valdez why his
marker is green. Neither one can answer, but luckily, graduate
student Abby Stanton has designed an experiment to help.
Click the URL below for more details.
https://youtu.be/uahUzob29go

Now, can you explain why most leaves are green?

Introduction

Give it a try!! Click the URL below for virtual paper


chromatography practice. Try select types of plant pigment extract
and types of solvent and get your own result.

http://www.amrita.olabs.edu.in/?sub=79&brch=17&sim=124&cnt=4

13
SB025 Pre-Lab Module

1. What are photosynthetic pigments?


________________________________________________________________________
Molecules that absorb specific wavelengths (energies) of light and reflect all others
________________________________________________________________________
to absorbs the photon for photosynthesis

2. State the main photosynthetic pigments in photosynthesis.


________________________________________________________________________
Chlorophyll a

3. Where are the photosynthetic pigments located in the chloroplast?


________________________________________________________________________
Thylakoid membrane and intergranal lamella

4. What is paper chromatography technique?


________________________________________________________________________
A technique for separating and identifying pigments and other molecules from cell extracts
that contain a complex mixture of molecules.
________________________________________________________________________

5. State TWO factors that cause different pigments to move at different rates.
________________________________________________________________________
1. attracted to different degrees to the fibres in the paper
________________________________________________________________________
2. molecular size

Experiment

1. State the variables for this experiment:


a. Manipulative : ________________________________________________________
Photosynthetic pigments
b. Responding : ________________________________________________________
Observed Rf values
c. Constant : ________________________________________________________

2. Why do you need to grind the leaves with acetone?


________________________________________________________________________

3. What is the function of solvent?


________________________________________________________________________

4. Why do we use the pencil instead of pen to mark the pigmented areas?
________________________________________________________________________

5. What is meant by Rf value?


________________________________________________________________________

14
SB025 Pre-Lab Module

Answer questions 6-9 using the information in FIGURE 1:

FIGURE 1

6. What is the distance from the origin to the final point of the solvent travelled?
________________________________________________________________________

7. Calculate the Rf value for pigment A-E.

Pigment Calculation Rf value


A

8. Based on the Rf value, determine which pigments:


i. Has the highest molecular weight : __________________________________
ii. Has the lowest molecular weight : __________________________________

15
SB025 Pre-Lab Module

9. What is the relationship between Rf value and different molecular weight of


photosynthetic pigments?
________________________________________________________________________
________________________________________________________________________

10. How do you make sure that the pigment spot (point of origin) is not immersed in the
solvent?
________________________________________________________________________
________________________________________________________________________

16

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