Name: ___________________________________ Yr.

& Sec: ___________________

Score: _____
Date: ___________________ Instructor ______________________________________

ANALYSIS OF SALIVA

THEORY
Saliva, like urine, is a good barometer of metabolic activity, since both are derived from
plasma. For this reason, analysis of the usual and unusual components of saliva may
help in the diagnosis of a pathologic condition in an individual.

MATERIALS
Saliva sample 0.5 N HCl
3 M HCl 1% Kl – starch solution
3 M NaOH 0.1M FeCl3
1 N HAc Ammonium molybdate
Acetone 5% AgNO3
Benedict’s solution Erlenmeyer flask (100 mL)
2.5 M NaOH Beaker, small
0.05% CuSO4 solution Funnel
Filter paper Evaporating dish
litmus 6 test tubes

PROCEDURES
I. Isolation of Mucin from Saliva
Rinse the mouth with water and collect 15 mL of saliva. Add 3 drops of 1 N
HAc and mix. To 30 mL of acetone, in an Erlenmeyer flask, add the saliva
slowly shaking the flask after each addition. Stopper the flask and set aside for
30 minutes. Filter and wash the precipitate formed, using two 3 mL portions
of acetone. Allow the precipitate to drain completely. Remove the filter paper

ANALYSIS OF SALIVA 1
 from the funnel, spread it out on a watch glass and allow the precipitate to
dry. Use the precipitated music to test for the possible components of saliva.

II. Qualitative Tests for the Compounds of Saliva

1. Carbohydrate
Place about ½ of the isolated music in a test tube. Add 1 mL of 3 M HCl and
immerse in a boiling water bath for 20 to 30 minutes. Cool and make faintly
basic with 3 M NaOH, then add 3 mL of Benedict’s solution. Heat again in
a boiling water bath until a visible change occurs.

Observation: ______________________________________________________________
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__________________________________________________________________________
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2. Protein
Dissolve the remaining portion in 1 mL of 2.5 M NaOH, then add three
drops of 0.05% CuSO4 solution. Mix.
Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
_________________________________________________________________________

3. Nitrate
Mix 1 mL of 0.05 N H2SO4 two drops of 1% Kl, and two drops of starch
solution. Add 2 mL of saliva. Mix.

Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
ANALYSIS OF SALIVA 2
 4. Thiocyanate
Two drops of saliva, add three drops of freshly prepared 0.1 M FeCl3 solution
and a few drops of 1 N HCl.

Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
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Smoke a cigarette outside the laboratory room and immediately after, collect another 2
mL sample of your saliva. Repeat the test for thiocyanate. (NOTE: cigarette smoke
contains traces of CN and it is believed that the presence of CNS is the byproduct of
detoxification mechanisms to rid the body of CN.)

Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
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Glucose
Using a saliva sample collected 15 minutes after eating, add 1 mL of Benedict’s
solution to 2 mL of saliva, then heat in a boiling water bath.

Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
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ANALYSIS OF SALIVA 3
 Inorganic Phosphate
Mix 1 mL of saliva and 1 mL of ammonium molybdate solution. Heat and
observe.

Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
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Chloride
Mix 1 mL of saliva with three to five drops of nitric acid followed by five
drops of AgNO3 solution.

Observation: ______________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
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What ion is present in saliva by above result?

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ANALYSIS OF SALIVA 4
Summarize your result in the table below.

Substances Test for Test Reagents Results Conclusions

Carbohydrate NaOH, Benedict’s

solution, heat

Protein NaOH, CuSO4

Nitrate H2SO4, Kl starch

CNS – FeCl3, HCl

Glucose Benedict’s reagent,

heat

Inorganic Phosphate Ammonium

molybdate

Chloride HNO3, AgNO3

ANALYSIS OF SALIVA 5