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Analysis of Saliva
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John Emmanwel Aljhu C. Del Rosario
Department of Biology, College of Science, PUP Sta. Mesa, Manila

ABSTRACT

Saliva which contains mucins, amylase and other proteins is a viscous fluid with a pH around 6.8.
In this experiment collection of three samples of saliva was done to test each of its components such as
amylase, mucins, calcium and inorganic phosphate and to determine if these are present in the samples.
Results (in the form of colored precipitate and changes in color of the solution) that were obtained were
all positive for each component of the saliva which indicates that these components are present in the
samples.

Key words: Saliva, mucins, amylase, calcium, inorganic phosphate

Introduction Methodology

Saliva contributes to the digestion of Collection of Saliva


food because it lubricates and allows the
swallowing of food more convenient. It contains The mouth was rinsed to get rid of food
enzyme such as amylase which is responsible for particles. About 10 ml of warm water was taken
breaking down of starch into simpler sugars that into mouth and moved it about by the tongue for
can be later absorbed by the small intestine. nearly a minute. Fluid was collected in a clean
Mucin is a glycoprotein which can be beaker. The tests were performed with this
precipitated by acetic acid at around pH 4.5 has diluted saliva.
a function in saliva. It provides protection in oral
Test for Amylase
activity. Calcium and inorganic phosphate are To 2.5 ml of starch solution in a test
also an essential component of the saliva that tube, 1 ml of 0.1M phosphate buffer, pH 7.0 was
can also be precipitated using acetic acid (for added followed by 1 ml of 0.1 M NaCl solution.
calcium) and concentrated nitric acid (for 1 ml of salivary fluid was also added to the
inorganic phosphate). mixture. Mixed. At intervals of one minute, one
Objective drop of the mixture was transfer to depressions
in a spot plate, containing a drop of iodine
The aim of this experiment is to test if solution.
there is a presence of amylase, mucins, calcium
and inorganic phosphate in the saliva which can Test for Mucins
be seen as colored precipitates. A 1% acetic acid was added from a 2mL
of salivary fluid drop by drop.
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Test for Calcium Saliva was rich in the enzyme amylase


To 2 ml of salivary fluids, 5 drops of 1% which is the main enzyme to breakdown starch.
acetic acid and 5 ml of 2% potassium oxalate Amylase hydrolyses 1,4 glycosidic bonds in
solution were added. starch at random. The break-down products do
Test for Inorganic Phosphate not have capacity to bind iodine. In this test
To 2 ml of salivary fluids, few drops of using iodine there is no large enzymatic reaction
concentrated nitric acid were added followed by happens in sample A and B because it results a
a pinch of ammonium molybdate. The solution black blue color. But there is a brown in C
was then warmed. which can determined small amount of
enzymatic reaction.
Results and Discussion
Test for Mucins
Test for Amylase

B
Figure 2.Test for mucins shows a thread-like
precipitate in the three samples of salivary
solution.

A thread-like precipitate was formed


which indicates the presence of mucins. Test
tube C has the longest thread-like precipitate
C
which indicates a greater amount of mucin is
present on it. Test tube A and B are the same.
Mucin is a glycoprotein constituent of mucus.
The precipitate was formed because of the acetic
acid which is around pH 4.5. Test of Mucins is
performed to know if there are any symptoms of
abnormalities that are happening to your body.
Figure 1.Test for amylase with different saliva Increased mucin production occurs in
sample. many adenocarcinomas, including cancers of the
pancreas, lung, breast, ovary, colon and other
tissues.
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Test for Calcium of inorganic phosphate present in the saliva.


Inorganic phosphate combined with ammonium
molybdate to form a yellow colored ammonium
phosphomolybdate. The precipitate ammonium
phosphomolybdate is
the inorganic salt of phosphomolybdic acid. It
contains the remarkable phosphomolybdate ion
complex.

Conclusion

These particular tests for the different


components of the saliva which gave a positive
Figure 3.Test for calcium shows a white in all test generally indicates that these
precipitate formed as calcium oxalate. components are always present in the saliva like
the amylase which in the experiment gives a
After the addition of acetic acid, reaction is black blue color, without the presence of it
evident. The previously slightly clear solution starch in the food we eat will never be break
turns white. Then after the potassium oxalate down because starch is necessary to break down
was added small granules of precipitate was into glucose monomer by amylase through
formed. Only, small amount of precipitate was hydrolyzing the 1,4 glycosidic bonds of starch
collected. A faint white precipitate was formed. to form glucose monomer. Positive results are in
Calcium ions are precipitated as calcium oxalate the form of colored precipitates like canary
under neutral or slightly acidic conditions. yellow colored ammonium phosphomolybdate
Test for Inorganic Phosphate in the test for inorganic phosphate while a white
thread-like precipitate was formed in the test for
mucins which indicates the presence of mucins.
All in all, analysis of saliva is necessary to
determine what substances are present in saliva.

References

Hayman, J.M Jr., and Johnston, Sara M.: The


Excretion of Inorganic Phosphate, J. Clin
investigation 11:607 (May) 1932

Power, M. H., and Wakefield, E. G.: A, Proc.


Staff Meet, Msyo Clin. 6: 401 (July 8) 1931
Figure 4.Test for inorganic phosphate shows a
canary yellow (ammonium phosphomolybdate) Cope CL. Determination of inorganic phosphate
precipitate. in human blood-plasma by micro-
titration. Biochem J. 1931;25(4):11831189.
In this experiment, test tube A and C has
the lesser precipitate formed while test tube B
Dodgson KS. Glycosulphatase: observations on
has the greater which indicates a greater amount
the activity of partially purified preparations
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towards the sulphate esters of certain


monosaccharides and steroids. Biochem J. 1961
Feb;78(2):324333

DODGSON KS, SPENCER B. Assay of


sulfatases. Methods Biochem Anal.1957;4:211
255.

McPherson RA, Ben-Ezra J. Basic examination


of saliva. In: McPherson RA, Pincus MR,
eds. Henry's Clinical Diagnosis and
Management by Laboratory Methods. 22nd ed.
Philadelphia, PA: Elsevier Saunders; 2011:chap
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