ANALYSIS OF SALIVA
Activity No. 9
I. PROCEDURE AND OBSERVATIONS:
1. Test for Mucin:
2 ml of saliva + 2 drops of dilute CH3COOH --------------- obs: white precipitates/
coagulates
What is the precipitate formed? Mucin, a glycoprotein which is covalently bonded with
carbohydrate.
What is its function? It functions as lubricant.
2. Test for Proteins:
2 ml of saliva + 3 drops of Biuret reagent -------------------------- obs: violet color
Describe the results. Violet solution is formed. It’s an indication for the presence of
protein in saliva.
3. Test for Calcium
5 ml K2C204 solution.
2 ml saliva + 5 drops of 1% CH3COOH ----------------------------------------
Describe the results : formation of white precipitates
Write the chemical equation.
HoAc
Ca+2 + K2C2O4 ----------------→ CaC2O4 ↓
4. Test for Inorganic Phosphate
3 drops of conc. HNO3
2 ml saliva + (NH4)2MoO4 ------------------------------------
WB
Describe the results: formation of yellow precipitates
Write the chemical equation.
HNO3
PO4-3 + (NH4)2MoO4 -----------------→ (NH4)3 PO4 ·12 MoO4 ↓
5. Digestion of Starch ( Hydrolysis of Starch)
Stir
25 ml of starch paste + 1 ml of saliva---------------------
Wb 40 0C
30 mins
At 1-minute intervals remove using a glass rod. A drop of the mixture and put it into a drop of
iodine solution in a spot plate. Observe the color changes.
Color 1st few minutes ---------------------- blue
_____________ purple
_____________ red
_____________ yellow
______________ colorless
At achromatic point:
wb
5ml of the solution + 5ml Benedict’s reagent --------------------------
Describe the results Orange colored precipitates
Explain. At this point, hydrolysis reaches to a slightly positive transformation from starch
to shorter sugar chains. Benedict’s reagent gives an orange color precipitate upon heating.
QUESTIONS:
1. What is the reaction with iodine when the Benedict’s test becomes positive?
At the achromatic point, iodine solution gives colorless solution. At the same time, giving
a slightly positive result with Benedict’s reagent, an indication of the conversion of starch to
achrodextrin.
2. What is the reduction of Benedict’s reagent due to?
Conversion of starch to glucose by salivary amylase.
3. How long did it take for a complete transformation of the starch into reducing sugar?
Starch beraks down to shorter chain sugar with enzyme salivary amylase. This process is
called hydrolysis. Salivary amylase is an endoamylase attacking the interior of the
polysaccharide chain on α 1-4 glycosidic linkage. Complete hydrolysis of starch requires the
presence of another enzyme maltase which acts on α 1-6 branch points.
4. Is it possible to get a reduction test before the achromatic point is reached?
Yes, when the fragmentation of starch reaches to erythrodextrin, Benedict’s reagent is
reduced giving an orange color precipitates upon heating.
5. What happens to starch when it is digested with saliva?
Starch when hydrolysed by saliva (amylase) is broken down into:
Amylose → amylodextrin → erythrodextrin → achrodextrin → maltose → glucose
6. What substances are responsible for the reaction?
Hydrolysis of starch (salivary digestion) is due to the presence of enzymes. Salivary
amylase commonly known as PTYALIN which is taking place in the buccal cavity and to
a certain extent in the fundic end of the stomach.
7. Would drinking too much liquid ( H20) during meals favor or disrupt digestion?
Drinking water during or after a meal actually aids digestion.
8. Is there any relationship between the pH of saliva and the susceptibility of dental carries?
Yes. Dental caries also known as tooth decay is the breakdown of teeth due to the
activities of bacteria. Acidic environment boosts batcerial growth.
9. Outline the different stages of Starch Digestion with Salivary amylase for both Iodine and
Benedict’s reagent.
Iodine sol’n Benedict’s reagent
Starch blue Negative (blue)
(insoluble in water
↓
Amylose blue Negative (blue)
(soluble in water)
↓
Amylodextrin purple negative (blue)
↓
Erythrodextrin red (+) very slightly positive (green)
↓
Achrodextrin colorless (++) slightly positive (orange)
↓
Maltose colorless (+++) positive (orange)
↓
Glucose colorless (++++) very positive (red orange)
