Fat Hydrolysis

Investigating the Hydrolysis of Fats and Oils
Background
Fats and oils are esters of triglycerols and fatty acids. Hydrolysis can break down
a fat or oil and release the triglycerol and fatty acids. The acids can be separated
and identified and this information can be used to identify the original fat or oil.
Practical Techniques
You will need to find out about volumetric analysis (titrations) and how to make
up accurate solutions. You also need to find out about using a pH meter
Where to start
An enzyme called lipase catalyses the hydrolysis of the fats and oils. When the
hydrolysis occurs the fatty acids will be released and the acidity of the reaction
mixture will rise. An alkali can be added to the reaction mixture to neutralise the
fatty acids. The rate at which the alkali needs to be added can be used as a
measure of the rate of the hydrolysis.
Plan an experiment to hydrolyse an oil and measure the rate of hydrolysis.
Possible Investigations
• Investigate the effect of changing the concentration of the enzyme on the

rate of the reaction.
• Investigate the effect of changing the concentration of the oil on the rate of
the reaction.
• Investigate the effect of changing the concentration and /or type of the
detergent on the rate of the reaction.
• Investigate the effect of changing the temperature on the rate of the

reaction.
• Investigate the effect of changing the type of oil on the rate of the reaction.
• Investigate the effect of changing the pH at which the experiment is run at.
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Sources of Information
• Thorpe A., Making a standard solution, Chemistry Review, November

2002
• Shipton M., Fats and Oils, Unilever Educational Booklet: Advanced Series,
• The Chemistry Video Consortium and The Royal Society of Chemistry

(2000) Practical Chemistry for Schools and Colleges CD ROM
• Ferguson M., Volumetric Analysis, Chemistry Review, September 1996
• Denby D., Investigating Enzymes, Chemistry Review,
• Lindsey D., Measuring pH, Chemistry Review, September 1998
• Vokins M. (ed), (1994), Nuffield Advanced Science Chemistry Students

Book, Longman (Third edition)
• Faust C.B., Jassal S.S., (1993) Lipids – a consumer’s guide, Education in

Chemistry, Vol 30,1
• Thorpe A., Assessing the risks in practical work, Chemistry Review,

September 2000
• Thorpe A., Experimental error and error analysis: just how good are those
results, Chemistry Review, November 2001
23
Teachers' Notes
General
The Unilever Fats and Oils Booklet is an excellent source for this
investigation and has full experimental details for the starter experiment.
Students can find it initially difficult to carry out the technique but with
practise, clear results can be obtained.
Each experiment can take up to 1 hour.
Chemical Principles
Esters, hydrolysis, kinetics
Essential Equipment
Burette, pH meter, magnetic stirrer
Essential Chemicals
sodium hydroxide, solid lipase, calcium chloride, sodium lauryl sulphate,

glycerol triethanoate (or triethanoyl glycerol or triacetin)
Safety
No risk assessment has been given. It is essential that students prepare a

detailed risk assessment before they start. Teachers must be satisfied that
this is suitable for the proposed investigation.
24
Starter Experiment Sheet - Investigating hydrolysis of fats and oils
A basic procedure is given below to measure the rate of hydrolysis is given

below.
Prepare the following solutions
• 0.01 mol dm-3 sodium hydroxide solution (placed in a burette)

• 0.5g solid lipase (the enzyme) in 10 cm3 of water – add 0.1g calcium
chloride to activate
• 5% sodium lauryl sulphate detergent
You will need to think about how much of each solution to prepare. This will
depend on how much of the solution is used in each experiment and how many
experiments you do (including any repeats).
You will also need glycerol triethanoate (or triethanoyl glycerol or triacetin) – this
is the oil that you will hydrolyse.
Take the enzyme solution and carefully adjust the pH to 8, using drops of the
sodium hydroxide solution.
Place equal volumes of the oil and the detergent solution into a beaker and
shake vigorously. This is now an emulsion of the oil in water. Carefully adjust the
pH to 8 using drops of the sodium hydroxide solution.
Place 20 cm3 of the emulsion and another 10 cm3 of the detergent into a beaker
and stir continuously with a magnetic stirrer.
Place a pH electrode into the mixture and measure the pH. Carefully adjust the
pH to 8 using drops of the sodium hydroxide solution.
Add 2 cm3 of water to the mixture and carefully adjust the pH to 8 using drops of
the sodium hydroxide solution and start a stop clock.
You are now ready to start taking readings. At regular time intervals add the
sodium hydroxide to re adjust the pH to 8. (It may be difficult to reach exactly 8
so you may have to approximate). Continue for about 5 minutes.
You now need to repeat the experiment by substituting 2 cm3 of enzyme for the
water.
You can plot a graph of volume of alkali added against time and the gradient will
give an initial rate of reaction in cm3 of 0.01 mol dm-3 of alkali per second. You
can convert this into moles of acid produced per second.
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Investigating the Hydrolysis of Fats and Oils

Plan an experiment to hydrolyse an oil and measure the rate of hydrolysis.

• Investigate the effect of changing the concentration of the enzyme on the

• Investigate the effect of changing the temperature on the rate of the

• Thorpe A., Making a standard solution, Chemistry Review, November

• The Chemistry Video Consortium and The Royal Society of Chemistry

• Ferguson M., Volumetric Analysis, Chemistry Review, September 1996

• Denby D., Investigating Enzymes, Chemistry Review,

• Lindsey D., Measuring pH, Chemistry Review, September 1998

• Vokins M. (ed), (1994), Nuffield Advanced Science Chemistry Students

• Faust C.B., Jassal S.S., (1993) Lipids – a consumer’s guide, Education in

• Thorpe A., Assessing the risks in practical work, Chemistry Review,

Esters, hydrolysis, kinetics

Burette, pH meter, magnetic stirrer

sodium hydroxide, solid lipase, calcium chloride, sodium lauryl sulphate,

No risk assessment has been given. It is essential that students prepare a

A basic procedure is given below to measure the rate of hydrolysis is given

Prepare the following solutions

• 0.01 mol dm-3 sodium hydroxide solution (placed in a burette)

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