Osazone formation

Introduction
• Osazones are formed when the sugars react with a compound known
as phenylhydrazine at boiling point.
• General Reaction:
• These sugars are reducing ones which have either a free aldehyde or
a ketone group to react with the phenylhydrazine.
• The reaction is stepwise
• Osazone formation involves hydrazone formation at C-1 of an aldose
(or C-2 of a ketose) and oxidation of C-2 (or C-1) of an alcohol group
to a ketone (or an aldehyde). The new carbonyl group is also
converted to a hydrazone.
• Osazone crystals have a characteristic shape under the light
microscope and help in the identification of the sugar type.
• Time to Form
• The time needed to create osazone crystals varies among the various
sugars involved, but helps to identify the sugars being tested. For an
osazone crystal to be presented from a hot solution will take as long
as follows:
 Title: Identification of unknown sugars by osazone formation
Apparatus:
• Test tubes
• Conical flask
• Spatula
• Measuring flask
• Gloves
• Funnel
• Water bath
• beaker
Chemicals:
• Carbohydrate (sugar)
• phenylhydrazine hydrochloride
• sodium acetate
• water
 Procedure
• Place 0.2g of carbohydrate,
• 0.4g of pure white phenylhydrazine hydrochloride (Caution: handle with great care),
• 0.6g of crystallized sodium acetate
• 4ml of water in a dry test tube(weigh the quantities with in accuracy 0.01g)
• stopper the tube loosely with a cork and put the solution in waterbath at 60-70C.
• Note the time of immersion and the time when osazone first separates. Shake the tube ocassionaly.(without
removing it from the boiling water) in order to prevent supersaturation. The precipitates separates quite
suddenly
• Note weather the precipitates is white crystalline or oily.
Result
• The product from mannose is simple hydrazone and is practically white. Arabinose osazone separates first as
an oil, while that from galactose is highly crystalline. Lactose and maltose give no precipitation from hot
solution.