BIOLOGY PRACTICAL RECORD WRITING

FOR CLASS XI
@

Educate  Enrich  Enlighten

PART 2

TOPIC
EXPERIMENTS 06 - 10

1
2
 06

PAPER CHROMATOGRAPHY
AIM: To separate and study the plant pigments by paper chromatography.

REQUIREMENTS

Fresh spinach leaves, chromatographic paper (Whatman No. 1), a

wide-long test tube, a split cork, mortar and pestle, petroleum ether,
acetone, funnel, beaker, filter paper, capillary tube sand etc.

PROCEDURE

1. Grind a few spinach leaves with little fine sand and about 5 ml
of acetone in a mortar and pestle. Filter it to get acetone extract

of the leaf pigments.

2. Take a narrow strip of chromatographic paper (Whatman No. 1).
Cut one end of the strip into a narrow notch.
3. Put a drop of the pigment extract in the middle of the strip near

the notch with the help of capillary tube. Allow the drop to dry
and repeat till four or five drops are placed on the paper.
4. Take the test tube and pour about 5 ml of ether-acetone solvent
(9 ether: 1 acetone) in it. Now hang the pigment extract loaded

chromatographic strip in the test tube with the help of a split cork,
in such a way that the loading spot lies about 1 cm above the sol-

vent level.
5. Make the cork air tight and place the test tube undisturbed for
some time, when solvent rises about 3/4th of the strip, take out the
strip carefully and let it dry.

3
CALCULATION
Rf value of each pigment spot can be calculated as follows:
Distance travelled by the compound
Rf =
Distance travelled by the solvent

Table : 8.1 Rf value of different plant pigments at room temperature

Pigments Colour of Spot Rf Value

1. Carotene Yellow Orange 0.95

2. Xanthophyll Yellow 0.71

3. Chlorophyll a Bulish green / dark green 0.65

4. Chlorophyll b Yellow with green 0.45

4
OBSERVATION
The dried chromatographic paper strip shows four distinct pigment

bands. Different leaf pigments can be identified by their colours.

Measure the distance of each pigment band from the loading spot
and also the distance travelled by the solvent.

RESULT:
The upper orange yellow corresponds to carotene .The Yellow hand
below it marks the Xanthophylls. The third from above dark green
represent chlorophyll a. The lower most yellowish green band is that
chlorophyll b.

PRECAUTIONS:

1. Spinach leaves should be fresh and green

2. The loading spot should be small and concentrated
3. The loading spot should be placed 2 – 3 cm away from the tip of
the notch.
4. While handling the strip in the test tube, the loading strip
should remain about 1cm above the solvent level.
5. The Chromatographic paper strip should not touch the walls of
the test tube.
6. The chromatographic paper strip should not be dried in sun
light.

5
6
 07

DISTRIBUTION OF STOMATA ON THE TWO SURFACES OF A LEAF

AIM : To study the distribution of stomata on upper and lower
surface of leaf and to calculate the stomatal index.

REQUIREMENT:
Fresh leaf of petunia or any other herbaceous plant/ fresh leaf of

lily, forceps, needle, blade, brush, watch glass, petridish, beaker,

glass slides, safranine , glycerine, water etc.

PROCEDURE :
1. Take out epidermal peels the upper and lower surfaces of
petunia leaf and put them in water in separate watch glasses
.The peels can be taken out by tearing the leaf obliquely with a
single jerk or scraping it with blade.
2. Stain the peels with Safranin by adding a few drops of the
stain in the watch glasses
3. Cut a small rectangle or square piece from peel and mount
them in glycerine on separate slides
4. Cover the peels with cover slip and observe them under the

microscope
5. Count the number of stomata and the number of epidermal cells
in the peels of both upper and lower epidermis of the leaf
appearing in the microscopic field .
6. Similarly prepare the slides of the peels of lily leaf and count the
number of stomata and the number of epidermal cells in the
microscopic field.

7
8
 7. Calculate the stomatal index of each surface of the leaf using
the following formula

No.ofstomata
Stomatal index := x100
No.ofstomata + No of epidermalcells.

OBSERVATION :
In the leaf of petunia, the number of stomata is more in the lower
epidermis and only few stomata are present in the upper epidermis.
However, the leaf of lily has almost equal number in both the epidermis.

PRECAUTIONS :
1. The curling of the peel should be avoided.
2. Always use brush to transfer the peel from watch glass to the slide
3. Excess of glycerine should be removed by using blotting paper.

9
10
 08

RATE OF RESPIRATION

AIM:
To study the rate of respiration in germinating seeds having different
substances such as wheat (carbohydrates) , mustard /ground nut
(fats) and gram / bean (proteins).

REQUIREMENTS:

A conical flask , cork, beaker, a twice bent glass tube , a small test
tube ,thread , water, KOH, germinating seeds of wheat, mustard /
groundnut and gram / bean , etc.

PROCEDURE :
1. Take a definite quantity (i.e 10gm) of germinating seeds of
wheat in the conical flask and hang a small test tube
containing KOH Crystal inside the flask with the help of a thread
2. Introduce one end of the bent test glass tube into the conical
flask through the cork. Dip the free end of the tube in a beaker
containing water.
3. Make the apparatus air tight and leave the apparatus
undisturbed.
4. Also set a control without germinating seeds in the flask.
5. Repeate the experiment taking the other types of germinating
seeds (mustard / groundnuts) and gram /bean.
6. Observe the level of rise of water in the glass tube dipped in water
for each type of seeds and record the data in the observation table.

11
OBSERVATION :

Sl.NO Type of germinating seeds

Rise in water level after 30 minutes’s

1.
Normal condition(Control)
No change in the water level

2.
Wheat Rise in the Water level

3.
Mustard / groundnuts Rise in the Water level

4.
Gram / Bean Rise in the Water level

12
RESULTS :

The raise in the water level is more in the case of wheat as

compared to mustard / groundnuts and gram / bean seeds .

CONCLUSION :
The germinating seeds respire and take oxygen from the flask and
release out carbon dioxide .The KOH crystals in the testtube absorb
carbon dioxide creating a vaccum in the flasks .This results rise of
water in the glass tube.
The rise in water level is comparatively less in the case of mustard /
groundnuts and gram /bean seeds because these seeds utilise fat and
proteins as respiratory substance and release less carbon dioxide .The rise
in water level is more in case of wheat grains as the later utilise
carbohydrate as respiratory substrate.

PRECAUTIONS :
1. The apparatus should be kept air tight.
2. Fresh KOH should be taken for the experiment.
3. The raise in water level should be accurately measured .

13
URINE TEST FOR SUGAR

14
 09

TEST FOR PRESENCE OF SUGAR IN URINE

AIM: Object To detect the presence of sugar (glucose) in urine.

REQUIREMENTS:
Urine sample (simulated prepared sample containing glucose
may be used), test tubes, test tube stand, spirit lamp, Benedict’s
solution, Fehling’s solution-A and Fehling’s solution-B.

PROCEDURE AND OBSERVATION

1. Benedict Test: Take 2ml of the urine sample in a test tube and

add 2ml of Benedict’s solution to it. Boil it and then cool it to

room temperature.
Green, yellow or brickred precipitate appears.
2. Fehling’s Test: Take 2ml of the urine sample in a test tube
and add 2ml of each of Fehling’s solution-A and Fehling’s
solution-B.
Boil for about for 2 minutes and then cool it to room
temperature.
Greenish yellow, yellowish red and reddish orange precipitate s
is formed.

CONCLUSION
1. In Benedict’s test, green precipitate shows traces, yellow
precipitate shows moderate and brick red precipitate shows
intense quantity of glucose in the urine sample.
2. In Fehling’s test, greenish yellow, yellowish red or redish orange
precipitates show the presence of traces, moderate and large
quantity of glucose respectively in the urine sample.

15
16
PRECAUTIONS
1. Wash the test tube thoroughly before use.
2. Use standard reagents.

17
URINE TEST FOR ALBUMIN

18
 10

TEST FOR PRESENCE OF ALBUMIN IN URINE

AIM: To detect the presence of albumin in urine.

REQUIREMENTS
Urine sample (simulated/prepared sample containing albumin
may be used), test tubes, test tube stand, spirit lamp, sulphosalicylic
acid, Robert solution (HNO3+MgSO4) and dropper.

PROCEDURE AND OBSERVATION

1. Sulphosalicylic Acid Test: Take 2ml of urine sample in a test
tube and add an equal volume of 30% sulphosalicylic acid to
it. Heat the tube gently.
A whitish or cloudy turbidity appears in the solution.
2. Heller’s Test: Take 5ml of Robert’s solution (HNO3+MgSO4) in a
test tube. Tilt the tube and add urine with the help of a dropper
on the inner side of the test tube so that it forms a layer over
the Robert’s solution.
A white ring is formed at the junction of Robert’s solution and
urine sample.

CONCLUSION
1. In sulphosalicylic acid test, the whitish/cloudy turbidity shows
the presence of albumin in the urine sample.
2. In Heller’s test, formation of a white ring is an indicative of
the presence of albumin in the urine sample.

19
20
PRECAUTIONS
1. Wash the test tube thoroughly before use.
2. Use standard reagents.

21
22