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Respiration

To prove the evolution of carbon dioxide during respiration.


Method: an apparatus is set up as shown in the figure . the stoppered u- shape tube contains soda
lime. The bottle B contains lime water. The bottle C contains well watered germinating seeds.
The bottle D also contains lime water.and is connected to an aspirator with a stop cork. The bottle
C is covered with a black cloth.
Observation: when the stop cork of the aspirator is opened, the lime water in D turns milky after
some time. This is because on opening the stop cork a partial vacuum is created and air is
drawnin slowly through the other end. The air passing through the sodalime is freed of co2 and
therefore does not turn the lime water in bottle B milky.
respirometer is an apparatus used for measuring the rate of respiration.
Respiroscope is an apparatus used to demonstate respiration.
To demonstrate aerobic respiration in germinating seeds.
Method: place a few germinating seeds in a respiroscope and invert it in the beaker containing
mercury as shown in the figure. With the help of forceps insert a stick of caustic potash into the
respiroscope. The whole setup should be air tight. Set up a similar control but withour any seeds.
Allow the whole apparatus to stand for a day.
Observation: the level of mercury in the respirometer is rises. in the control, the level of the
mercury stays the same. It is obvious that the oxygen inside the respiroscope has been taken in by
the plant during respiration which lowers the air pressure in the apparatus. So the mercury rises.

To prove that heat is generated during respiration:


Method: take two thermos flasks. In the other flasks take some germinating seed that have been
killed by boiling for a few minutes. Soak these sead seed in formaldehyde to prevent any heat
generation through fermentation. Both the flasks are fitted with air tight corks through which are
inserted thermometers. The bulbs of the thermometers are kept embedded within the muss of
seeds. The initial temperature is noted in either case.
Observation: after a few hours it is observed that while no rise in temperature is is observed in the
case of seeds that have been killed by boiling, there is distinct rise in temperature in the case of
germinating seeds. The rise in temperature is obviously due to the liberation of the heat energy by
the respiring seeds.
How was the heat produced in flask A?
Ans. aerobic respiration.
Why it was important to sterilize the surfaces of both set of peas?
Ans. Sterilizing ensure killing of all microbes. Microbes can do anaerobic respiration too. So to
prevent the extra heat production it is important to do sterilization.
What is the function of cotton wool?
Ans. Cotton wool closes the flasks . it lets gases pass in and out of the flask.
Why the tharmoflask kept inverted?

Ans. to let the co2 pass easily. Co2 is lighter than normal air. So it can pass through the cotton
wool easily. Otherwise excess co2 can kill the seeds.
An investigation into the effect of temperature on the activity of amylase
The digestive enzyme amylase breaks down starch intosugar maltose. If the speed at which the
starch disappears is recorded, this is a measure of the activity of the amylase.
The figure shown apparatus which can be used to record how quickly the starch is used up.
Spots of iodine are placed into the depressions on the spotting tile. 5 cm3 of starch suspension is
placed in one boiling tube, using a syringe, and 5 cm3 of amylase solution in another tube, using
different syringe. The beaker is filled with water at room temperature. Both boiling tubes are
placed in the beaker of water for 5 minutes, and the temperature recorded.
The amylase solution is then poured into the starch suspention, leaving the tube containing the
mixture in the water bath. Immediately , a small sample of the mixture is removed from the tube
with a pipette and added to the first drop of iodine on the spoting tile. The colour of the iodine is
recorded.

Anaerobic respiration in yeast


Yeast is a micro organism which, like your muscles, can respire in two ways. You can
investigate some aspects of this in a simple experiment.

The indicator Janus Green B changes colour according to the amount of oxygen present.
Oxygen present
Indicator oxidised to blue colour

Oxygen absent
Indicator reduced to pink colour

Method
1
Place 20cm3 of glucose solution with some yeast added to in a test tube. Add two
drops of indicator solution. The colour produced will tell you whether oxygen is
in the mixture.
2
Pour enough liquid paraffin over the mixture to form a layer right over the
surface. This layer will prevent more oxygen from the air getting into the
mixture.
3
As soon as the colour of the mixture indicates that there is no oxygen present, fix
up a delivery tube leading to a small amount of bicarbonate/indicator in another
test tube.
4
Design and set up a suitable control. Remember that in this experiment you are
trying to find out whether the yeast is respiring in the absence of oxygen.
5
Leave the apparatus for 10-20 minutes and then record your observations.

Questions
1

What can you conclude from the final colours of?

2
3

(a)
the bicarbonate indicator solution?
(b)
The Janus Green Bb indicator solution?
What can you conclude from your two answers to question 1?
Why is a layer of liquid paraffin used in the experiment?

If yeast is left in a sugary liquid for some days, a characteristic smell is produced.
This is the result of ethanol accumulating in the mixture. Fermentation is the
name given to this process in which yeast turns sugar into ethanol:
Sugar
ethanol
+
carbon dioxide
Suggest a way of discovering whether any energy is released during this process.

experiments on respiration
An equation summarizing the process of aerobic respiration is given below.
C6H12O6

6O2

6CO2

6H2O

Energy

glucose

(decrease
in dry weight)

uptake of oxygen

output of carbon dioxide

e.g. heat production

Respiration in living organisms


Study the Figure on p.5.02 to form a clear idea of the design of the apparatus but do not
assemble it yet.
(a) Fill a beaker or jar two thirds full with cold water and place it on a tripod and gauze.
(b) Label two boiling-tubes A and B
(c) Place a known weight of germinating seeds in boiling-tube A and an equal weight of
dead seeds (killed by boiling) into tube B and then push a plug of cotton wool into each
tube.
(d) Put about 5 g soda-Iime on top of the cotton wool in each tube and place the tubes in
the water bath.
(e) If the manometer does not already contain liquid, use a dropping pipette to half fill the
reservoir of the manometer with coloured liquid. Air bubbles should be removed by
gently blowing and sucking down the long arm of the manometer.
(f) Check that the screw clips are open and fit the bungs and manometers securely into
the tubes.
(g) Move the rubber marker on the manometers to indicate the liquid level.
(h) Leave the test-tubes in the water bath for a few minutes to acquire the temperature of
the water
(i)

Close the screw clips and note any change in the level of liquid in the
manometer over a period of 10 to 20 minutes.

Discussion

I What change in the liquid level did you observe in the two tubes ?
2 What does this change tell you about the volume of air in each boiling tube ?
3 What are the principal gases in the atmosphere and what is the percentage volume of
each ?
4 What is the function of the soda-lime in this experiment? What might happen if it were
not included?
5 Which atmospheric gases could be responsible for the change in the volume of air in
the boiling tube?
6 Say why you regard your results as reasonable evidence that respiration is taking place
in the organisms under investigation.
7 Why were the two tubes placed in a water bath?
8 Tube B is a 'control' to the experiment. What is its purpose?

5 Discussion answers
I In tube A the liquid should rise. In tube B there should be little or no change.
2 A rise in level indicates that there is a reduction in the volume of air in tube A.
3 Air contains approximately 20% oxygen, 79% nitrogen and 0.04% carbon dioxide.
4 The soda lime absorbs any carbon dioxide produced by the living material. If the living
organism produced as much carbon dioxide as it consumed oxygen, there would be no
change in volume unless the carbon dioxide were absorbed.
5 The absorption of the tiny amount of carbon dioxide from the air will make no
perceptible difference to the manometer. Thus the decrease in volume could be due to
removal of oxygen, nitrogen or both. Since nitrogen does not usually participate in living
reactions, the most likely cause of the diminution in volume is the removal of oxygen.
6 Aerobic respiration involves the production of carbon dioxide and the absorption of
oxygen. Since the soda lime absorbs any carbon dioxide given out, the reduction in
volume is consistent with the hypothesis that the organisms are using up oxygen.
7 Any change in temperature of the tubes will cause the air inside them to expand or
contract and so affect the manometer. The water is less susceptible to temperature
fluctuations and will maintain a fairly constant temperature throughout the experiment.
8 The control tube (a) shows up any change in volume due to temperature change,
(b) shows that removal of oxygen is the result of living activities rather than simple
physical absorption of oxygen by the material.

Experiment 12 The effect of temperature on fermentation rate


You are provided with a boiling tube containing a suspension of yeast in a solution of
glucose. DO NOT SHAKE THE TUBE at any stage of the experiment.
(a) Set up a clamp stand, clamp, tripod, gauze and water bath as shown in Fig. 1
(p.12.02). Clamp the boiling tube securely in the water bath.
(b) Fit the manometer and syringe assembly as shown in the diagram ensuring
(i) that the bung makes an air-tight seal in the mouth of test- tube and
(ii) the tap is open (turned upwards).
(c) Use a dropping pipette to fill the reservoir of the manometer about three quarters full
of coloured liquid. If the liquid does not flow into the capillary, close the tap (horizontal)
and raise the syringe plunger. Air bubbles in the column can be expelled by using the
syringe to push the
liquid and trapped air back to the reservoir.
(d) Turn the tap downwards, push the syringe plunger to the bottom of the barrel and
open the
tap (vertically upwards). Move the marker on the capillary tube to indicate the level of
liquid
and then copy the table on p. 12.02 into your notebook..
(e) Record the temperature of the water bath, note the time and close the tap (horizontal).
Leave the tap closed for one minute and keep withdrawing the syringe plunger to bring
the liquid back to the marker. After one minute bring the liquid level exactly back to the
mark, open the tap (upwards) and record in your table the volume indicated on the
syringe.
This figure represents the volume of carbon dioxide given out by the yeast in one minute.

(f) Turn the tap downwards, push the syringe plunger to the bottom and close the tap
(horizontal).
(g) Take two more readings of the volume of gas produced in one minute and calculate
the average of the three.
(h) Check that the tap is open (upwards) and use a small Bunsen flame to raise the
temperature of the water bath by about 5 C. Leave the apparatus for two minutes after
heating has stopped.
Record the new temperature and take three more one-minute readings as before, but this
time
record the temperature at the end of the three readings to obtain the mean temperature
during
the experiment.
(i) Repeat the operation from (g) raising the temperature about 5 C each time until you
reach 45
or 50 C. At temperatures between 30-40 C the three readings should be made in rapid
succession to avoid volume changes due to cooling; at 40-50 c you should add 0.1 cm3
to all readings to compensate for the cooling effect.

Discussion
1 In general terms what was the effect of rising temperature on the rate of carbon dioxide
production?
2 Was the change in rate consistent over the whole of the temperature range which you
tried? If
not, suggest an explanation for any deviation.
3 As the water bath cooled down during each series of three readings, there would be a
contraction in the volume of gas. How could this affect the accuracy of your readings?
Suggest a control which could be employed to correct any errors of this kind.
4 The solubility of gases in water decreases with increasing temperature. How might this
property affect your results?
5 What factors, apart from temperature, might be limiting the rate of carbon dioxide
production
from yeast cells?

Discussion - answers
1 In general, a rise in temperature is accompanied by an increase in carbon dioxide
production
and the relationship is linear, at least up to 35 oC.
2 At temperatures beyond 40 oC the rate of carbon dioxide production will level out and
then start to decrease. This may be due at first to limiting factors, e.g. the rate of intake of
substrate, and finally to increasing numbers of yeast cells being killed by the high
temperature (thermal death point).
3 The effect of cooling would be to make the readings at the higher temperatures too low,
since the decrease in volume would counteract the increase due to gas production.
A suitable control would be an identical respirometer in the water bath but containing 10
cm3 of glucose solution only. The decrease in volume due to cooling would be measured
by the syringe,
and its volume added to the reading in the experimental tube.
4 The reduced solubility of carbon dioxide at higher temperatures would result in the
expulsion of some dissolved gas with each increase of 5 oC. This might make the first
reading of each group of three slightly higher than the others.
5 Limiting factors are likely to be the availability of the substrate in solution, the rate at
which it
can be taken up by the yeast cells, the availability of enzymes, co-enzymes etc. in the
cell, the rate at which the carbon dioxide can be expelled from the cell and possibly the
accumulation of inhibitory metabolites.

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