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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
enzyme such as amylase which is responsible for
breaking down of starch into simpler sugars that
can be later absorbed by the small intestine.
Mucin is a glycoprotein which can be
precipitated by acetic acid at around pH 4.5 has
a function in saliva. It provides protection in oral
activity. Calcium and inorganic phosphate are
also an essential component of the saliva that
can also be precipitated using acetic acid (for
calcium) and concentrated nitric acid (for
inorganic phosphate).
Objective
The aim of this experiment is to test if
there is a presence of amylase, mucins, calcium
and inorganic phosphate in the saliva which can
be seen as colored precipitates.
The mouth was rinsed to get rid of food
particles. About 10 ml of warm water was taken
into mouth and moved it about by the tongue for
nearly a minute. Fluid was collected in a clean
beaker. The tests were performed with this
diluted saliva.
Test for Amylase
To 2.5 ml of starch solution in a test
tube, 1 ml of 0.1M phosphate buffer, pH 7.0 was
added followed by 1 ml of 0.1 M NaCl solution.
1 ml of salivary fluid was also added to the
mixture. Mixed. At intervals of one minute, one
drop of the mixture was transfer to depressions
in a spot plate, containing a drop of iodine
solution.
Test for Mucins
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%
acetic acid and 5 ml of 2% potassium oxalate
solution were added.
Test for Inorganic Phosphate
To 2 ml of salivary fluids, few drops of
concentrated nitric acid were added followed by
a pinch of ammonium molybdate. The solution
was then warmed.
Results and Discussion
Test for Amylase
which is the main enzyme to breakdown starch.
Amylase hydrolyses α 1,4 glycosidic bonds in
starch at random. The break-down products do
not have capacity to bind iodine. In this test
using iodine there is no large enzymatic reaction
happens in sample A and B because it results a
black blue color. But there is a brown in C
which can determined small amount of
enzymatic reaction.
Test for Mucins

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.

Test for Calcium
Figure 3.Test for calcium shows a white
precipitate formed as calcium oxalate.
After the addition of acetic acid, reaction is
evident. The previously slightly clear solution
turns white. Then after the potassium oxalate
was added small granules of precipitate was
formed. Only, small amount of precipitate was
collected. A faint white precipitate was formed.
Calcium ions are precipitated as calcium oxalate
under neutral or slightly acidic conditions.
Test for Inorganic Phosphate
Figure 4.Test for inorganic phosphate shows a
canary yellow (ammonium phosphomolybdate)
precipitate.
In this experiment, test tube A and C has
the lesser precipitate formed while test tube B
has the greater which indicates a greater amount
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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
in all test generally indicates that these
components are always present in the saliva like
the amylase which in the experiment gives a
black blue color, without the presence of it
starch in the food we eat will never be break
down because starch is necessary to break down
into glucose monomer by amylase through
hydrolyzing the α 1,4 glycosidic bonds of starch
to form glucose monomer. Positive results are in
the form of colored precipitates like canary
yellow colored ammonium phosphomolybdate
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
Cope CL. Determination of inorganic phosphate
in human blood-plasma by micro-
titration. Biochem J. 1931;25(4):1183–1189.
Dodgson KS. Glycosulphatase: observations on
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):324–333

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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