APPROVAL SHEET

The complete report of practicum plant physiology with title “Measurement of

chlorophyll content”, created by:

Name : Nur Fahmi Utami

ID : 101 404 155

Class : Biology Bilingual

Group : V

it has been checked and consulted to Assistant/ Assistant Coordinator shall be

accepted.

Makassar, Mei 2nd 2012

Assistant Coordinator, Assistant,

Risna Irawati , S.Pd Yusmar Yusuf NIM . 081404172

Known by,

Lecturer of Responsibility

Drs. Ismail, M.S NIP . 196112311986031015

CHAPTER IINTRODUCTION

A. Background All living things either directly or indirectly require a result of

photosynthesis in which plants. Because the photosynthetic process is capable of

converting light energy into chemical energy in the form of organic compounds.

Photosynthesis the process itself is a process of formation of organic material

from inorganic ingredients (co2, h2o, h2s) with the help of the sun's light energy

and the major pigment of chlorophyll contained in chloroplasts. In addition to

chlorophyll, other pigments are also involved some of which are carotenoids,

which serves to form chlorophyll in the light harvesting and protect chlorophyll

from photo oxidation process. The process of photosynthesis produces

photosyntat form of glucose (c6h12o6) is needed by other living things.

The relation between chlorophyll content and rate of photosynthesis that

is data are presented which support the conclusion of emerson that the rate of

photosynthesis is proportional to the chlorophyll content when the latter is varied

by varying the iron supply. These data give a straight line passing through the

origin, which is not true of emerson's results, similar data are presented which

show that a similar relation exists when nitrogen controls the chlorophyll content,

evidence is given which indicates that magnesium plays a part in the process of

photosynthesis in addition to its effect upon the chlorophyll content.

Most of the chlorophyll present in leaves of the chlorophyll can also be

found in parts of green plants such as roots, stems, fruit, and flower seeds in

limited amounts. Although most of the chlorophyll is distributed in the leaves,

but not spreading evenly. The amount of chlorophyll in the leaves will be

different from the base of the tip, middle and both sides of the leaf. Besides

differences in leaf color also shows the difference in the amount of chlorophyll.

The green color is highly correlated with leaf chlorophyll content. In general, the

higher the green color of leaf chlorophyll content. In this experiment evaluated

the content of chlorophyll in leaves of different ages in plants.

B. The PurposeTo determine of chlorophyl degree a, b and total chlorophyll of leaves.

C. The Benefit

Students University can determine levels of chlorophyll from a plant when they

are doing the observation.

CHAPTER IIPREVIEW OF LITERATURE

Photosynthesis is the process by which plants, some bacteria, and some

protistants use the energy from sunlight to produce sugar, which cellular respiration

converts into ATP, the "fuel" used by all living things. The conversion of unusable

sunlight energy into usable chemical energy is associated with the actions of the

green pigment chlorophyll. Most of the time, the photosynthetic process uses water

and release the oxygen that we absolutely must have to stay alive (Anonymousa,

2012).

Energy source of all Living Things is the sun. Solar radiation reaching the

earth is only a small portion of the electromagnetic spectrum. Wavelengths of

sunlight That Reaches the earth's surface covers 310 to 2300 nm. Wavelength of 225

nm (ultraviolet) is also irradiated by the sun, but λ is very high photon energies that

are harmful to many lives, was blocked by a layer of ozone in the atmosphere at the

top. Rays are longer than 2500 nm (2.5 μ) is mainly eliminated by water vapor and

CO2 in the atmosphere. Visible light is the fraction of light That Reaches the earth

(Sasmitamihardja, 1996).

That absorbs molecule to visible light is colored or black pigment. Electron

into an excited electron car usually associated with unsaturated double bond. Eg

chlorophyll, have a high level of unsaturation and efficiently absorbs light,

particularly blue and red light (Sasmitamihardja, 1996).

Pigment is an object that can absorb light. Color pigment visible light that is

reflected (in other words, not absorbed). Chlorophyll, the green pigment in

photosynthetic cells, absorbs all visible light except green, which is reflected and

captured by the eye. Black pigment absorbs all visible light. White pigment reflects

all the colors (Ismail, 2009).

Chlorophyll a measurements have historically provided a useful estimate of

algal biomass and its spatial and temporal variability. The fluorometric method is

extensively used for the quantitative analysis of chlorophyll a and phaeopigments.

However, errors can be introduced into the results when chlorophylls b and/or

chlorophylls c are present. Chlorophyll b is the main source of error in this method.

While generally not abundant in surface waters, chlorophyll b can be as high as 0.5

times the chlorophyll a concentration in the deep chlorophyll maximum, causing

slight underestimations of the chlorophyll a concentration, and drastic

overestimations of the phaeopigment concentrations. Divinylchlorophyll a also

interferes and is taken as chlorophyll a by this method. The procedure described here

is appropriate for all levels of chlorophyll a concentration in the marine environment.

Filtration volumes should be modified for the different environments. Scientists who

employ this or other methods to measure pigments should make themselves aware of

the current and historical issues that surround these techniques and make appropriate

decisions about specific methodologies for their application based on the scientific

requirements and constraints of their individual programs (Anonymousb, 2012).

The more concentrated a solution of a colored substance, the more absorbing

light, so it looks darker, the relationship between the absorption of light by the

solution concentration, is the basic principle of usability spectrophotometer. The

concentration of a solution of colored substances may also be known easily, based on

the price of absorbance (OD = Optical Density), because concentrations are linearly

related to OD. In addition, by using Spectrophotometer spectronic 21 D can also

directly measured the concentration of a solution (Ismail, 2012).

The light has two properties, namely the nature of the wave and particle

properties. Properties of light particles are usually expressed in photons and quanta,

which is a packet of energy that has its own characteristics: each photon has a

wavelength Certain. Energy in each photon is inversely proportional to wavelength,

so the wavelength of violet and blue light has a higher photon energy than the

wavelength of light orange (orange) and red (Sasmitamihardja, 1996).

CHAPTER IIIPRACTICUM METHOD

A. Date and Place

Day/Date : Thursday, April 11th 2012

Time : At 10.50 – 13.00 pm

Place : Biology Laboratory of right side in 3rd floor at FMIPA UNM

B. Tools and Materials

1. Tool

a. Spectrophotometer

b. Large test tube

c. Measuring pipette

d. Mortar

e. Funnel

f. Cuvet

2. Materials

a. The young leaves and old leaves of plants Hibiscus rosa-sinensis

b. Distilled water

c. Filter paper

d. Ethanol

e. Aluminum foil

C. Work Procedure :

1. Weighted 0.5 grams of fresh leaves then grind with a mortar until smooth.

2. Dissolved with acetone 80% as much as 10 ml.

3. Sift into a large test tube, if necessary volume to 10 ml both ends meet and

cover with aluminum foil.

4. Measured the absorbance of the extract using a spectrophotometer at λ 645

and 663 nm.

5. Calculated the levels of chlorophyll

CHAPTER IVOBSERVATION RESULT AND DISCUSSION

A. Observation Result

No. The species of plant AbsorbanceThe color of leaf

Young leaf Old leaf

1. Hibiscus rosa-sinensis

649 0,532 1,190

665 1,560 1,400

Data analysisCount of chlorophyll degree of following the formula Holden (1965) as follows:Chlorophyll a = 0,0127 A663 - 0,00269 A645 (mg ml-1)Chlorophyll b = 0,0229 A645 - 0,00468 A663 (mg ml-1)Total chlorophyl = 0,0202 A645 + 0,00802 A663 (mg ml-1)When using 96% ethanol solvent used Wintermens formula and de Mots (1965) as follows:Chlorophyl a = 13,7 A665 - 5,76 A649 (mg l-1)Chlorophyl b = 25,8 A649 – 7,6 A665 (mg l-1)Total chlorophyl = 20,0 A649 + 6,1 A665 (mg l-1)

1. Young leafChlorophyl a = 13,7 A665 - 5,76 A649 (mg l-1)

= 13,7 (1,560) – 5,76 (0,532)= 21,372 – 3,06432= 18,30768 (mg/l)

Chlorophyl b = 25,8 A649 – 7,6 A665 (mg l-1)= 25,8 (0,532) – 7,6 (1,560)= 13,7256 – 11,856= 1,8696 (mg/l)

Total chlorophyl = 20,0 A649 + 6,1 A665 (mg l-1)= 20,0 (0,532) – 6,1 (1,560)= 10,64 – 9,516= 1,124 (mg/l)

2. Old leafChlorophyl a = 13,7 A665 - 5,76 A649 (mg l-1)

= 13,7 (1,400) – 5,76 (1,190)= 19,18 – 6.8544= 12,3256 (mg/l)

Chlorophyl b = 25,8 A649 – 7,6 A665 (mg l-1)= 25,8 (1,190) – 7,6 (1,400)= 30,702 – 10,64

= 20,062 (mg/l)Total chlorophyl = 20,0 A649 + 6,1 A665 (mg l-1)

= 20,0 (1,190) – 6,1 (1,400)= 23,8 – 8,54= 15, 26 (mg/l)

B. DiscussionIn Experiments measuring the chlorophyll content, we use the young

leaves and old leaves from different plants. In general, the color of the leaves

on the material Artocarpus heterophylla & Hibiscus rosa-sinensis is the

which contained green chlorophyll a and chlorophyll b. chlorophyll content

of leaves has owned by the difference in the analysis of data. In the data

analysis, the which has a lot of the content of chlorophyll in green young

leaves where chlorophyll content is 18.30768 (mg / l), while the dark green

leaves contain little chlorophyll That, namely 15, 26 (mg / l).

Based on research of data showed the highest chlorophyll a levels in

young leaves. While the lowest levels of chlorophyll a on old leaves. For

chlorophyll b levels in young and old leaves do not have much difference

and can be said nearly equal. Difference Between the levels of chlorophyll a

to chlorophyll b levels are on average each plant contains more chlorophyll b

than chlorophyll a. Whereas for determination of total chlorophyll content

was highest in old leaves and the lowest found in young leaves. In other

words we can make-initial Conclusions that the chlorophyll levels will be

higher with increased age of the leaves.

Based on the data analysis and graph above shows the higher levels

chlorophyl that by accretion or leaf age. The green color is very closely

related to leaf chlorophyll content. In general, the older leaves of the green

leaf color will be higher chlorophyll content. In Addition to the structure and

metabolism have been more perfect when old leaves compared with young

leaves in photosynthesis is high and the effect on protein synthesis. This is

the first indicator that shows, that the older age of a leaf, the higher levels of

chlorophyll they contain.

In this experiment are consistent with the theory Because the light

green leaves contain the most chlorophyll compared with the dark green

leaves that most of the leaves of the plants have chlorophyll, but in different

amounts depending on the type of leaves. Green leaves contain more

chlorophyll than the leaves of another color (Anonymous, 2012).

CHAPTER VCONCLUSION AND SUGGESTION

A. ConclusionBased on the observation that we do can be concluded to measure chlorophyll

content a, b and chlorophyl total at leaves hibiscus rosa-sinensis that young and

old, we use formula of wintermens and mots (1965) that usde the 96% ethanol

solution. Which is young leaves on chlorophyll a: 18.30768 (mg / l), chlorophyll

b: 1.8696 (mg / l), and its chlorophyl total: 1.124 (mg / l). The old leaves,

chlorophyll a :: 12, 3256 (mg / l), chlorophyll b: 20.062 (mg / l), and its total

chlorophyll: 15, 26 (mg / l).

B. Suggestion

1. Suggestion for the laboratory;

Students are expected to be more careful and meticulous in performing lab

work as needed to measure the chlorophyll content of high-precision money.

2. Suggestions for assistant.

Assistants are expected to always control the student for this lab in desperate

need of a high degree of accuracy.

3. Suggestions for Laborant staff

Laboratory should always check laboratory equipment in order for lab work

if needed, will always be available

BIBLIOGRAPHY

Anonymousa. 2012. Photosynthesis, (Online), http://www. biobookps.html, access

April 17th 2012.

Anonymousb. 2012. Chapter 14. Measurement of Chlorophyll a and Phaeopigments

by Fluorometric Analysis , (Online), http://ocean.stanford.edu/cal

/JGOFS_chla.pdf, access April 17th 2012.

Ismail. 2009. Fisiologi Tumbuhan. Makassar: Jurusan Biologi FMIPA UNM.

Ismail & Abd. Muis. 2012. Penuntun Praktikum Fisiologi Tumbuhan. Makassar:

Laboratorium Biologi FMIPA UNM.

Sasmitamihardja, dradjat. 1996. Fisiologi tumbuhan. Bandung : FMIPA ITB.