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Osazone Formation

1. This experiment identifies unknown sugars through osazone formation, a reaction where sugars react with phenylhydrazine at high temperatures to form characteristic osazone crystals. 2. Sugars containing an aldehyde or ketone group will undergo this reaction. The procedure involves mixing the sugar with phenylhydrazine hydrochloride and sodium acetate, then heating and observing the formation and properties of any precipitates. 3. Two example sugars tested were arabinose, forming an oily precipitate in 9 minutes, and galactose, forming white crystalline precipitates in 5.5 minutes, identifying them based on reaction time and crystal properties.

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1K views

Osazone Formation

1. This experiment identifies unknown sugars through osazone formation, a reaction where sugars react with phenylhydrazine at high temperatures to form characteristic osazone crystals. 2. Sugars containing an aldehyde or ketone group will undergo this reaction. The procedure involves mixing the sugar with phenylhydrazine hydrochloride and sodium acetate, then heating and observing the formation and properties of any precipitates. 3. Two example sugars tested were arabinose, forming an oily precipitate in 9 minutes, and galactose, forming white crystalline precipitates in 5.5 minutes, identifying them based on reaction time and crystal properties.

Uploaded by

inam ullah
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Title

Identification of Unknown Sugars by Osazone Formation


Principle
In this experiment, a monosaccharide reacts with phenyl hydrazine which is a crystalline
compound. The sugars which will reduce as a result of this reaction will result in the formation
of osazones.

Apparatus
1. Beakers
2. Conical Flask
3. Spatula
4. Measuring Flask
5. Gloves
6. Funnel
7. Water Bath
8. Test Tube
Chemicals used
1. Water
2. Carbohydrate (sugar)
3. Phenylhydrazine hydrochloride
4. Sodium Acetate
HSE related to Chemicals
1. Carbohydrate: A biomolecule consisting of C, H and O. Most common source of energy
for most organisms. Build macromolecules and spare protein and fat for other uses. Bad
carbohydrates are rapidly digested and increases the risk of heart diseases, obesity and
diabetes.
2. Phenylhydrazine hydrochloride: Molecular formula is C6H9CIN2. Used to form
phenylhydrazones of natural mixtures of simple sugars. Widely used reagent in
conjunction with aldehydes and ketones. Breathing it can irritate nose, throat and lungs
causing coughing or shortness of breath. May cause a skin allergy.
3. Sodium Acetate: An organic sodium salt with formula CH3COONa. Used as additives in
food industry, concrete manufacture, heating pads and in buffer solutions. Inhalation may
cause a cough and sore throat. Direct skin or eye contact may cause redness and irritation.
Theory
Osazones are formed when sugars react with a compound known as phenylhydrazine at boiling
point.

The sugars which react are the ones with either a free aldehyde or a ketone group. Osazone
formation involves hydrazine formation at C-1 of an aldose and oxidation of C-2 of an alcohol
group to a ketone. The new carbonyl group is also converted to a hydrazone. Osazone crystals
have a characteristic shape under light microscope and help in identification of sugar type.
Procedure
1. Place 0.2 g of carbohydrate, 0.4g of pure white phenylhydrazine hydrochloride and 0.6g
of crystallized sodium acetate in test tubes.
2. Add 4mL of water in a dry test tube.
3. Stopper the test tube loosely with a cork and put the solution in water bath at 60-70 °C.
4. Note the time of immersion and the time when osazone first separates.
5. Shake the tube occasionally in order to prevent super-saturation.
6. Note whether the precipitates is white crystalline or oily.
Precautions
1. Wear personal protective equipment
2. Handle chemicals with care.
3. Always wear a mask.
4. Use gloves while heating test tubes.
Result and Conclusion
Test tube A is arabinose osazone which separates first as an oil and takes approximately around 9
minute. While test tube B is galactose which separates out as white crystals and takes around 5.5
minute.

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