General Biology 1

INTRODUCTION TO PHOTOSYNTHESIS

PLANT PIGMENTS
IMPORTANCE OF CHOLOROPHYLL AND OTHER PIGMENTS

Sources:
 Photosynthetic Pigments (http://biology4alevel.blogspot.com)

 Fun-Leaf-Chromotography-Activity (https://www.teachengineering.org)

Quarter 2 Week 2 Leo Jr Icutan and Isabelo Ramos

Quick Review!

How do Plants
Make Food through
Photosynthesis?
Fast Talk!
Fill the Missing Terms
Energy from the _____ drives all the Earth’s food chains.
Green plants trap _____ energy and use this to produce
__________ from the reaction of __________ and
__________ . This energy is then passed onto consumers.

The pigment which absorbs sun’s energy for photosynthesis

is called __________. All food production whether in plant
or animal depends on the process of photosynthesis.
Light Energy in Photosynthesis

When light strikes a

leaf, there are

3 possible
outcomes
 Light Energy in Photosynthesis
Reflected
12%

White light hitting a

leaf (or any surface) Absorbed
will be either:
83%

Of the light absorbed;

 4% is used for photosynthesis Transmitted
 Remaining 79% is lost as heat energy
5%
Light Energy in Photosynthesis

Light is a form of electromagnetic radiation which travels

in waves.
Wavelength

Light wave

The distance between two crests on a wave is called the

wavelength and is measured in nanometres (nm). 1
nanometer is 10-9 m)
Light Energy in Photosynthesis

Spectrum of light
 Visible light is made up of a
continuous sequence of
different colours.

 When a beam of light passes

through a prism the VISIBLE
spectrum of light is produced.
 Light Energy in Photosynthesis

Spectrum of light Most people

cannot see below
400nm or above
700nm (visible
part of the
spectrum).

Each colour of light has a different wavelength.

Light Energy in Photosynthesis

The color that we see is the

color that has been reflected

If a color has been absorbed

it appears black to the human
eye
 Spectrum

Light Energy
in Photosynthesis

The image on the

right shows that most
of the blue and red
wavelengths have
been absorbed,
which is why it
appeared black.
 Photosynthetic Pigments

 A pigment is any substance that absorbs light.

 When pigments absorb light, electrons are temporarily

boosted to a higher energy level.

Photo Credits:
biology4alevel.blogspot.com
 Photosynthetic Pigments

 are pigments presented in chloroplasts or photosynthetic

bacteria.
 They capture light energy necessary for photosynthesis
and convert it to chemical energy.
 Chloroplasts contain several different pigments, which
absorb different wavelengths of light.
 Photosynthetic Pigments

The photosynthetic pigments of

higher plants form 2 groups:
the chlorophylls and
the carotenoids.
 Photosynthetic Pigments

Chlorophyll is the main pigment that

absorbs mainly red & blue-violet
light, and reflect green light -
giving green leaves their color.
Photosynthetic Pigments

Photo Credits: http://biology4alevel.blogspot.com

 Photosynthetic Pigments

The reflected wavelengths are mainly green which is why

chlorophyll appears green to the eye.

Spectrum
 Photosynthetic Pigments

Carotenoids are orange pigments that

protect chlorophyll from damage by the
formation of single oxygen atoms (free
radicals).

 They can also absorb wavelengths of light

that chlorophyll cannot absorb, and pass
on some of the energy from the light to
chlorophyll.
 Photosynthetic Pigments

Carotenoids absorb strongly in the blue-

violet range. They are usually masked by
the green chlorophylls.

There are 2 types of carotenoid:

 carotenes (β-carotene)
 xantophylls.
Photosynthetic Pigments

Photo Credits: http://biology4alevel.blogspot.com

 Photosynthetic Pigments

Main pigment

Chlorophyll a is the most abundant

pigment in most plants. Its absorption
peaks are 430nm (blue) and 662nm
(red). It emits an electron when it
absorbs light.
 Photosynthetic Pigments

Accessory pigments

 Chlorophyll b is similar to chlorophyll a, but its absorption

peaks are 453nm and 642nm. It has a similar role to
chlorophyll a, but is not as abundant.
 Carotenoids : carotene and Xanthophylls.

Note: The combination of all of the pigments increases the range of

colors that plants can use in photosynthesis.
Absorption Spectra
a graph showing the percentage of
light absorbed by pigments, for each
wavelength of light.
Chlorophyll a and b: complex
molecules containing magnesium
which absorb light energy in
red/blue regions which is used for
photosynthesis.
Carotene & xanthophyll: absorb
light mainly in green part of the
spectrum and pass on the energy
to chlorophyll.
Action Spectra
Rate of photosynthesis
at each wavelength of
light.

The higher the rate,

the more effective the
wavelength of light at
providing energy for
photosynthesis
Action Spectra
Question
If you are a pigment,
what wavelengths of
light are you going to
absorb for higher
rate of
photosynthesis?
Comparing
Absorption &
Action spectra
This shows the
absorption spectrum for
Chlorophyll only…what
evidence is there that
other pigments are
involved in
photosynthesis?
 Questions to Ponder!
29

Can you now explain the importance of

plant pigments? Try answering these
questions:
1. Why are chlorophyll pigments important?
2. As regards accessory pigments, what roles
do they play if they are not directly
involved in photosynthesis?
ENRICHMENT ACTIVITY
Leaf Chromatography
 Pre-Assessment Questions
1. What is a solution?
 A solution is a mixture made of two or more substances.

2. What is a mixture?
 A mixture is created when two or more substances are
mixed but not chemically joined together.
3. What is photosynthesis?
 Photosynthesis is the process in which a plant uses light
energy from the sun to produce food they need to
survive.
Pre-Assessment Questions
4. What are pigments?
 Pigments are the natural color of matter in plant tissue or
animals.
5. What is chlorophyll and where can it be found?
 Chlorophyll is a compound found in cells of plants that absorb
sunlight to begin photosynthesis.
6. Why do leaves change colors?
 Leaves change color due to the chlorophyll breaking down in
the fall, which causes the other pigments that have been there
to begin to appear.
CHROMATOGRAPHY
 Chromatography is a method of separating
mixtures into its individual components.

 It works by absorbing the pigments and by

pulling the solvent up the paper.

 As the solvent moves up the paper it

separates the mixture into its colors or
components.

Source: Autumn Jewels

CHROMATOGRAPHY
We will explore about chromatography
with leaves and will learn about the
hidden colors or pigments in leaves!

The pigments are extracted from the

plant and separated on chromatography
paper.

The more soluble the pigment, the

further it will travel up the paper.
Source: Autumn Jewels
 EXTRACTION OF PIGMENT

1. Grind plant tissue with sand & propanone (acetone)

 Break open cells & release pigment
 Propanone to dissolve pigment

2. Filter mixture
 Remove sand & debris
 SEPARATION OF PIGMENT

3. Repeatedly spot extract onto

chromatography paper.
 Ensure sufficient mixture is present

4. Spot above solvent level

 Prevents pigment dissolving out of
paper.
Acetone or
alcohol
 SEPARATION OF PIGMENTS

5. Run chromatogram in acetone of

isopropyl alcohol
 Separates pigments as pigments
have different solubilities in this
solvent

6. Ensure paper does not touch sides

 Ensure solvent moves evenly up Acetone or
paper alcohol
 PHOTOSYNTHETIC PIGMENTS

Solvent Level (Final)

Most soluble
Carotene ( Orange )
Carotenoids
Xanthophyll ( Yellow )

Chlorophyll a ( Blue-green )
Chlorophyll b ( Dark green )
Least soluble
Spot Line
Solvent Level (Initial)
Pigment spot
 CALCULATING Rf
From the chromatogram,
we can identify the
different pigments based
on how far they have run.
This gives us each
pigments Rf value.
 CALCULATING Rf
 Rf stands for retardation or retention factor. You
can record the Rf value by knowing the distance
analyte travelled by the distance solvent travelled.

 Rf value is independent of the sample concentration.

Its value is always positive.
 CALCULATING Rf
From this, we can identify the
pigment by looking at known Rf
values.

Formula to use:

Rf = Distance travelled by pigment

Total distance solvent travelled
 CALCULTING Rf
Distance travelled by pigment
Rf =
Total distance solvent travelled

Red pigment
7.5/10 = 0.75

Blue pigment
2.5/10 = 0.25
 CALCULATING Rf
 1. Light green spot indicates chlorophyll-b pigment.
 Rfvalue= Distance chlorophyll-b travelled / Distance solvent travelled =
2/10 = 0.2

 2. Dark green spot represents chlorophyll-a pigment.

 Rfvalue= Distance chlorophyll-a travelled / Distance solvent travelled =
3.7/10 = 0.37
 CALCULATING Rf
 3. The yellow band represents xanthophyll pigment.
 Rfvalue= Distance xanthophyll travelled / Distance solvent travelled
= 5.6/10 = 0.56

 4. The yellow-orange band indicates carotene pigment.

 Rfvalue= Distance carotene travelled / Distance solvent travelled =
9/10 = 0.9
 CALCULATING Rf
 Factors affecting the Rf values of a particular analyte are:
 Stationaryphase
 Temperature

 Mobile phase

 The concentration of the mobile phase

 The concentration of the stationary phase

 CALCULATING Rf
 The Rf value of compounds in the mixture differs by any
changes in the concentration of stationary and mobile
phases.

 Temperature affects the solvent capillary movement and the

analyte’s solubility in the solvent.
Leaf Chromatography Summary
Preparation of Concentrated Leaf Extract
SEPARATION OF PIGMENTS
 Group Tasks
51

 Considering our enrichment activity:

Bring the following:
 leaves of 2 types of plants (green
and multi-colored)
 Denatured alcohol
 Ruler and observation notes
 Executethe steps stated in the
previous slides GAME TIME
 End of the Presentation
52 That’s for now. I hope you enjoyed our
discussions and activities. I hope you
keep in mind and at heart the points we
can learn from the generous plants, and
be like them if we can! 1 Timothy 6:18

by Leo Olivar Icutan Jr.