Hydrolysis of Starch by Salivary Amylase - PAPC

  

Biochemistry and Clinical Pathology -Practical PAPC

Aim: To study hydrolysis of Starch from salivary amylase.

Reference:

1. Rao. G. Devela. ‘A Manual of Practical Biochemistry’. Birla Publication. PVT LTD. Delhi. Second Edition. 2008. 04-10.
2. Chaudhari M.R. et.al. ‘Biochemistry and Clinical Pathology’. Nirali Prakashan. Pune. Thirty Fourth Edition. 2018. 3.1 – 3.14.
3. Indian Pharmacopeia. ' Ministry of Health and Family Welfare'. Indian Pharmacopeia Commission. Gaziabad. Vol I. 888.

Requirement:
Chemical: Strach, Salivary amylase diluted, 0.1N Iodine Solution, Acetic Acid, Hydrochloric Acid, Sodium Hydroxide, Phosphate buffer 6.8 pH, Distill Water etc.
Apparatus: Test Tube, Beaker, Water Bath, Watch Glass, Test Tube Holder, Glass Rod.

Principle:
Enzyme are responsible to regulate rate of reaction and Amylase break down Starch to simple sugar. On digestion of starch chracterstic inference will not be observed.

Procedure:
> Take 4 test tube with 1 ml of 1 % Starch solution and add 1 ml of Diluted salivary amylase into it.
> Add 1 ml of phosphate buffer into each test tube.
> Put them at different temperature as 0° C, 25° C, 37° C and 95° C for few minutes.
>  After that add 1 ml of salivary amylase to each and wait for a minute.
> 1 drop of mixture to watch glass and add one drop of Iodine  and observe the colour.

Observation Table:
S. No.Test TubeObservationInference
1.    I
Very light blue colourPartial digestion
2.II
No change in colourComplete digestion
3.
4.		III
IV		No change in colour
Very light blue colour 		Complete digestion
Partial digestion

Draw the following:


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Result: Effect of Salivary Amylase on hydrolysis of Starch was studied.

Saliva digest Strach at 25oC and 37oC degree centigrade temperature, but 0oC and 97o C diminished the activity of Enzyme.


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Summary

Carbohydrates are substance that possess a general formula CnH2nOn.

Test for carbohydrates can be performed by preparation of 1% solution. 

Hydrolysis:

Hydrolysis is uncontrolled process can be done by addition of HCl conc. and after that netralize by Sod. Carbonate with reference to litmus paper.

Different Reagents used in identification are as:

1. Molisch Reagent

2. Barfoed Reagent

3. Benedict Reagent

4. Scliwanoff Reagnet

5. Fehling A and B and other

Steps included are:

Molisch Test àIodine Test àBenedict Test à Barfoed Test àScliwanoff Test and àOsazone Test

Colours Produced are:

> Violet colour ring between two junction confirms Carbohydate presence.

> Blue and Reddish brown colour indicate the presence of starch or glycogen respectively by Iodine test and in case of no colour mono or di saccharide may be present.

> Reducing sugar shows Green yellow or Red colour precipitate with Benedict's Test.

> A brick red ppt indicate mono-saccharide presence with Barfoed Reagent.

> Scliwanoff reagent produces cherry red colour i.e. positive for keto sugar.

> Osazone Crystals

- Needle Shape à Glucose/ Fructose

- Sunflower Shape à Maltose

- Cottonball Shape à Lactose

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