Practical 2: Determination of the number of acetyl groups in an acetylated sugar

Lecturer: Ms Mokhosi

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 PLAGIARISM
The module outline provides information about plagiarism (under “Warning”), and further
instructions will be provided during lectures and practical's. Any form of copying or cheating will
not be tolerated in this module and will be dealt with according to the University guidelines.
Copying or cheating is a form of fraud, which is a criminal offence, and it does not help you in the
long term, and has very serious effects on the reputation of our University, and therefore the
reputation of your degree. We will take strong action against any student caught cheating or
copying, and will make every effort to ensure that cheating students are caught and prosecuted.
Several students in Semester 1 have been sent to the proctor and will face the consequences of
their behaviour.

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 Background
What is acetylation
 Acetylation- chemical reaction in which an acetyl group is added to a
compound
 Acetyl group- chemical fuctional group composed of a carbonyl group (C=O)
and a methyl group (CH3)

Determination of acetylated sugar

 Generally, an acetylated compound is hydrolysed with a standard amount of
alkali and the excess alkali is back-titrated with a standard acid.
 however, this approach is NOT USED with acetylated sugars, because the
liberated sugar undergoes slight resinfication by the present alkali, resulting
to a brown solution, with end point of titration difficult to detect accurately.

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Cont…
Acid hydrolysis
 Acetylated sugars hydrolysed with 1N H2SO4 instead of alkali,
resulting to rapid hydrolysis with clear solution
 Liberated acetic acid and excess h2so4 is back titrated with 1N
NaOH using phenolphthalein as an indicator.

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METHODOLOGY AND REAGANTS

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 KNOW YOUR REAGENTS

1. Acetylated sugar
2. Phenolpthalein (indicator)
3. 1N Sulfuric acid -direct contact with the eyes can cause permanent
blindness. Ingestion may cause internal burns, irreversible organ
damage, and possibly death.Respiratory tract irritation if inhaled
4. 1N Sodium hydroxide (NaOH)- severe burns (skin contact), vomiting,
nausea, diarrhea, etc (ingestion)
2-3 drops

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 Method
1. Label two 250 ml conical flasks A and B.
2. In A, place 1.5 g of the given acetylated sugar and 25 ml of 1N H2SO4.
3. In B place only 25 ml of 1N H2SO4.
4. Attach an air-condenser, add some boiling chips and reflux A gently for
30-40 minutes (until sugar dissolves).
5. Wash down the condenser with 10 ml distilled water.
6. Add 10 ml of distilled water to flask B to make the same vol.
7. Cool and titrate flasks A and B with 1 N NaOH using phenolphthalein as
8-10 an indicator. A faint pink color denotes the end point of the titration.
boiling
chips

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 Calculations and Results
Mass of unknown acetylated sugar is ? g

Flask A requires x ml 1N NaOH

Flask B requires y ml 1N NaOH
Difference in volume = (x-y) ml
 
1000 ml of 1N NaOH is equivalent to 1 acetyl group.
 
Therefore (x-y) ml of 1N NaOH ≡ (x-y) ml acetyl groups
1000
Therefore ? g of unknown acetylated sugar contains: (x-y) ml acetyl groups
1000

Therefore the unknown acetylated sugar has: (x-y) X mw of acetylated sugar

1000 ?g
  ≡ ________ acetyl groups
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Knowns…
1. Acetylated sugar has
already been weighed out
into vials (1.5g).

2. Molar mass of acetylated

sugar = 390.34 g/mol

3. Transfer all the sugar from

the vial into the conical
flask – rinse out the vial
with a little distilled water if
necessary (2-3 ml)

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