Lactose - Sugar PAPC

  

Biochemistry and Clinical Pathology -Practical PAPC

Aim: To perform the qualitative identification test for given sample of Carbohydrate.

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.

Requirement:
Chemical: Carbohydrate Testing Reagent, Sulphuric Acid, 0.1N Iodine Solution, Acetic Acid, Phenyl Hydrazine, Sodium Nitrate, Hydrochloric Acid, Sodium Hydroxide, Distill Water.
Apparatus: Test Tube, Beaker, Water Bath, Test Tube Holder, Glass Rod.

Observation Table:
S. No.ExperimentsObservationInference
1.Molisch's TestViolet colour ring between two junctionCarbohydrate Present
2.Iodine TestNo change in colourMono/ Di saccharide Present
3.Benedict’s TestGreen yellow colour seenReducing Sugar Present
4.Barfoed’s TestNo characterstic changeDi-saccharide Present


5.Osazone Testpuff-shaped crystalsLactose confirmed
(Draw the structure of Lactose at blank side.)

Result: On the basis of above observation it may found that the given sample of Carbohydrate was Lactose.


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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. 

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 gylcogen 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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