Experiment 1

RESPIRATORY VOLUMES
The lungs are always full of air. During the process of breathing
replenishment of fresh air. The total capacity of the lungs consists
named according to location and function. In order to understand the
it is necessary to be familiar with the various respiratory volumes.
be measured readily, as will be seen in this experiment.

there is a periodic
of various volumes
respiratory process,
Some of these can

Materials
1. Spirometer:
a. Autospiro-AS 500
b. Hutchinson Spirometer
2. Nose clips
3. Mouth piece
4. Alcohol 70% solution
5. Records the age, gender, height and weight of the subject. And records the
temperature of the room, humidity and the room air pressure.

Procedure and observations with Hutchinson Spirometer

1.

Clean up the mouthpiece of spirometer with ethyl alcohol and set the pointer at zero.

2. Tidal Volume

Tidal volume is the amount of air moved into and out of the lungs during natural
respiration. With the pointer set at zero mm, breathing as usual (inhale and exhale),
place the spirometer mouthpiece in the mouth, and exhale. Repeat the experiment
three times and records the volume.
3. Inspiratory Capacity

Inspiratory capacity is the maximum amount of air that can be inspired from the
resting expiratory position. With the pointer set at zero mm., inhale as deeply as
possible, place the spirometer mouthpiece in the mouth, hold the nose with nose clips
and exhale normally. Repeat the experiment three times and records the volume.
4. Inspiratory Reserve Volume (IRV)

IRV is the maximum amount of gas that can be inspired from the end-inspiratory
position. IRV equals Inspiratory capacity minus Tidal volume.
5. Expiratory Reserve Volume (ERV)

ERV is the maximum volume of air that can be expired from the end-expiratory

position. With the pointer set at zero mm., exhale normally, place the spirometer
mouthpiece in the mouth, hold the nose with nose clips and exhale as deeply as
possible. Repeat the experiment three times and records the volume.
6. Vital capacity.
Vital capacity is defined as the amount of air that can be exhaled forcibly after the
most forceful inspiration. With the pointer set at zero mm., inhale as deeply as
possible, place the spirometer mouthpiece in the mouth, hold the nose with nose
clips and exhale with maximum effort. Repeat the experiment three times and records the
volume.
Experiment 2

SALIVA AND SALIVARY DIGESTION

Saliva is produced by the three pairs of salivary glands: the submaxillary, the
sublingual, and the parotid. The amount of saliva secreted per day, in man, varies from
1 to 2 L. Normally only a minimal amount is produced until food is placed in the mouth.
Secretion of saliva is simple reflex phenomenon, but salivation can be conditioned; i.e.,
the sight, smell, or even thought of food can produce a copious secretion. Dehydration of
the body and emotional stress inhibit the production of saliva. The volume and character
of the salivary secretion depend upon the relative activity of the different glands and
upon the nature of the autonomic stimulation, parasympathetic or sympathetic.
Something of the character and composition of saliva is illustrated in the following
procedures.
Materials
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.

Litmus paper (pHydrion paper)

Dilute solution of acetic acid
Potassium oxalate solution
1 N HCL
1 N NaOH
1% starch solution
Iodine solution
Benedict's solution
Clean paraffin
Test tubes
Beakers
Porcelain spot plates
37C water bath.
Boiling water bath

Procedure and observations

1. Rinse out your mouth with several washes of distilled water; then chew a piece of

clean paraffin for a few minutes to stimulate the secretion of saliva. Collect the saliva
in a large test tube. Test the reaction (pH) of the freshly secreted saliva. Note the
acidity and viscosity of the saliva in the test tube.
1. Transfer about 5 ml saliva to a clean test tube and then add a few drops of acetic acid
solution. Observe and name the precipitate. Pour the saliva from one tube to another. Note the
viscosity.
2. To 2 ml saliva in a clean test tube, add 1 or 2 drops HCI. Heat the tube in a boiling

water bath for at least 10 min. Neutralize with 1 or 2 drops NaOH and then test for
reducing sugar by adding 10 ml Benedict's reagent and heating for a few minutes. Note the
color changes.
2. Place 25 ml I% starch solution into a beaker. Add 10 ml saliva and stir thoroughly.
After 3 minutes remove a small sample from the beaker and place it into one of the
spot plate depressions. Add to the sample 1-drop iodine solution and observe the blue color.
Repeat at 1-minute intervals until the iodine test for starch is negative. Next, place 5 ml
of the starch-saliva solution into a clean test tube and add 10 ml Benedict's solution. Note the
color changes.
5. Repeat the above experiment (no 4), using saliva that had been previously heated
to the temperature of boiling water for 10 minutes.
6. Test for the presence of calcium salts in freshly collected saliva by adding a few
drops acetic acid and a few drops potassium oxalate to 5 ml saliva.
Result
1. Acidity of the saliva (pH)
Viscosity of the saliva
viscous/liquid
2. Precipitation
+/Viscosity of the saliva
viscous/liquid
3. Color changes
Initial color
Final color
4. The iodine test for starch is negative after ........... Minutes
Color changes (Benedict's reaction)
a. Initial color
Final color
b. Initial color
Final color
5. The iodine test for starch is negative still positive.
Color changes (Benedict's reaction)

a. Initial color

Final color
b. Initial color
Final color
6. Calcium salts + /Conclusion
Experiment 3

DETECTION OF hCG IN URINE

The immunological detection of human chorionic gonadotropin (hCG) is now universally accepted
as the presumptive diagnostic test for pregnancy. The hCG is secreted by the developing placenta
and its appearance in urine can be detected as early as 7 days after conception, doubling in
concentration about every 1 .5 days, thereafter reaching a peak of 100-350 IU/ml by the end of the
first trimester.
Materials
I . Kit for direct latex agglutination test for detection f hCG in urine.
2. Urine samples : i.
Pregnant
ii. Non pregnant
Principles of Test
This kit is a direct latex agglutination test. Polystyrene latex particles have been optimally
coated with purified anti-hCG antibodies. The antibodies are beta chain specific, significantly reducing
the likelihood of false positive results due to LH, FSH and TSH at physiological levels. In the presence
of urine containing hCG an agglutination reaction occurs. Agglutination is considered a positive
result. The sensitivity of this kit is 0.2 IU hCG/ml, a level usually achieved 2-3 days after the first
missed menstrual period.
Procedure and observation
1.
2.
3.
4.

Place one drop of those urine samples on to the reaction area of the slide using a clean disposable
pipette/stirrer.
Shake the latex reagent, then using the dropper provided, add one drop of latex reagent and
mix using the flat end of the plastic pipette/stirrer, used in step 1.
Gently and evenly rock and rotate the test slide for 2 minutes whilst examining the test slide for
agglutination. Read the test at 2 minutes.
At 2 minutes the test should be examined in good light at a normal reading distance. The
appearance of agglutination pattern is indicative of the presence of hCG.

Result.

Urine sample :1. ...............................

2................................
CONCLUSION