Pharmaceutical Chemistry III

PHCMp 773

Oil Analysis (Part 1)

Practical Session 6

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Intended Learning outcomes By
By the end of this session, the students should be able to:  
a. Knowledge and Understanding
a.1 Recognize the basic principles of pharmaceutical definitions in terms of fats/oils and acid, saponification and ester
values, official pharmacopoeia methods of control and qualitative tests and handling complete monographs.
a.2 Explain the general pharmacopoeia statements, methods of performing quantitative tests and fat analysis.
a.4 Explain the importance of and the principles of various techniques in the determination of oil samples quality.
b. Professional and Practical skills
b.1 Use the proper pharmaceutical terms and abbreviations related to the official methods of control.
b.2 Practice handling and disposing chemicals, samples and oil preparations safely, according to GLP guidelines.
b.4 Apply techniques used to accurately evaluate the quality of an oil sample using acid value, saponification value and
ester value.
b.5 Judge the quality of a specific pharmaceutical/oil sample based on the results obtained and compose an official
report to communicate the results and conclusions made.  
c. Intellectual skills
c.1 Apply GLP (Good Laboratory Practice) guidelines while working in the lab.
c.3 Analyze and interpret observed experimental results to judge the oils’ purity.  
d. General and transferrable skills
d.1 Communicate clearly by verbal means through laboratory work and submitting a laboratory report.
d.2 Retrieve and evaluate quality information from experimental results from students' work.
d.3 Work effectively individually and in a team.
d.6 Implement documentation skills through submitting a lab report including interpretation of results of his\her oil
experiments.
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 Introduction
Why do we need Fat/oil Analysis?
Fats/oils are raw materials used in many pharmaceutical preparations e.g.
emulsions, creams, ointments & bases of suppositories.
So their quality needs to be checked.

Fats/Oils are:
Esters of 3 fatty acids and Alcohol (glycerol).

Fats (solid) are esters of SATURATED FAs while Oils (liquid) are esters of
UNSATURATED FAs.

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There are FIVE standard values to assess quality of oils. THREE of them are
discussed this lab.

1. Acid Value
2. Saponification Value
3. Ester Value
4. Iodine Value
5. Peroxide Value

4
 Acid Value

Definition:
The acid value IA is the number of mg of potassium hydroxide required to
neutralize the free acid in 1 g of the substance.
O O

R OH + KO H R O - +K + H2 O

Significance:
High acid value indicates high hydrolysis of ester back to acid and alcohol
and thus low oil quality.
This may be attributed to degradation by moisture due to bad storage.

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 Acid Value
General Method

Unless otherwise specified in the monograph, weigh 10 g (or mg) of the

substance being examined and add 50 ml of a mixture of equal volumes of
ethanol (96%) and ether R that has been neutralized with 0.1M potassium
hydroxide VS using 0.5 ml of phenolphthalein solution R as indicator. When
the substance has completely dissolved, titrate with 0.1M potassium
hydroxide, shaking constantly until a pink color that persists for at least 15
seconds is produced (n ml of 0.1 M KOH).
Calculate the acid value from the expression

IA = 5.610 n/m

Where n is the volume, in ml, of potassium hydroxide solution required and

m is the weight, in g, of substance taken.

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 Acid Value
Determination of the Acid Value for virgin linseed oil sample

Procedure
Dissolve 5.0 g of the linseed oil sample in 50 ml of neutral alcohol R ether R
mixture. 4.95-5.05 g
Titrate with 0.1M potassium hydroxide, using 2 ml phenolphthalein solution
R1 as an indicator.
Shake constantly until a pink color that persists for at least 15 seconds is
produced.
Calculate the acid value from the expression
IA = 5.610 n/m
where n is the volume, in ml, of potassium hydroxide solution required and m is the
weight, in g, of substance taken.

For virgin linseed oil, the acid value is < 4.5

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 Acid Value
Determination of the Acid Value for virgin linseed oil sample

Washing
Flask washed well with soap and
tap water and rinsed with distilled
water

Drying
Flask dried by little amount of ethanol

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 Acid Value
Determination of the Acid Value for virgin linseed oil sample

Weighing the Sample

Accepted range of error ?! 4.95-5.05 g

 Make sure all drops transferred to bottom
of flask and none hangs on the walls !!!
 Record the actual weight to use in
calculations.

5.0 g oil

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 Acid Value
Determination of the Acid Value for virgin linseed oil sample

The role of (alcohol/ether mixture)

 Ether is an organic solvent that dissolves

the oil.
 Alcohol is a co-solvent that allows
miscibility / homogenity between organic
layer (oil in ether) & aqueous layer (aq. KOH +
H2O produced in reaction).

This results in:

50 ml ethanol/ether
(1) allowing titration and completion of
mixture
acid base reaction.
5.0 g oil
(2) avoiding turbidity that might hinder
end point visualization.
Shake well 10
 Acid Value
Determination of the Acid Value for virgin linseed oil sample

Titrant is 0.1 M AQUEOUS KOH

End point is first persistent pink

Type of titration
Direct Acid Base titration

2 ml ph.ph Blank?
50 ml ethanol/ether Everything except the oil
mixture
5.0 g oil

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There are FIVE standard values to assess quality of oils. THREE of them are
discussed this lab.

1. Acid Value
2. Saponification Value
3. Ester Value
4. Iodine Value
5. Peroxide Value

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 Saponification Value
Ester value + Acid value

Definition:
The saponification value IS is the number of mg of potassium hydroxide
required to neutralize the free acids and to saponify the esters in 1 g of
the substance.

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 Saponification Value
General Method

Introduce the prescribed quantity of the substance to be examined (mg)

into a 250 ml glass flask fitted with a reflux condenser. Add 25.0 ml of
0.5 M alcoholic potassium hydroxide and boil under a reflux condenser
for 30 minutes. Unless otherwise prescribed, add 1 ml of
phenolphthalein solution R and titrate immediately (while still hot) with
0.5M hydrochloric acid (n1) Repeat the operation without the substance
being examined (n2).
Calculate the saponification value from the expression

Is = 28.05 (n2-n1)/m

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 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample
Procedure

Introduce 1.0 g of virgin linseed oil into a 250 ml glass flask fitted with a reflux
condenser. 0.95-1.05 g
Add 25.0 ml of 0.5 M alcoholic potassium hydroxide and boil under a reflux
condenser for 30 minutes.
Add 1 ml of phenolphthalein solution R1 and titrate immediately (while still hot)
with 0.5 M hydrochloric acid (n1)
Repeat the operation without the substance being examined (n2).
Calculate the saponification value from the expression
Is = 28.05 (n2-n1)/m
where n1 is the volume, in ml, of potassium hydroxide solution required, n2 is the volume,
in ml, of the blank , 28.05 is the equivalent factor and m is the weight, in g, of substance
taken.
For virgin linseed oil, the saponification value is 188-195 15
 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample

HCl

25.0 ml 0.5M alc. KOH

1.0 g oil

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 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample

Accepted
range of
error ?!
0.95-1.05 g

1.0 g oil

Sample Blank 17
 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample

Bulb pipette !!

25.0 ml 0.5M alc. KOH

1.0 g oil 25.0 ml 0.5M alc. KOH

Sample Blank 18
 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample

Boil on hot plate

(150 oC) for 30
minutes.

If flasks dried up,

add ethanol

25.0 ml 0.5M alc. KOH

1.0 g oil 25.0 ml 0.5M alc. KOH

Sample Blank 19
 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample

1 ml ph.ph
Excess un-reacted alc. KOH 1 ml ph.ph

R-COO-K+ + R’OH ≤ 25 ml 0.5 M alc. KOH

Sample Blank 20
 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample

Titrant is 0.5 M HCl.

End point is colorless.

Type of titration
Back Acid Base
titration

1 ml ph.ph
Excess un-reacted alc. KOH 1 ml ph.ph

R-COO-K+ + R’OH ≤ 25 ml 0.5 M alc. KOH

Sample Blank 21
 Saponification Value
Determination of the Saponification Value for virgin linseed oil sample
Why do we need a Back titration?
Very Slow Reaction ( 30 min)
Need heat (boiling)

Why do we use alcoholic KOH in the Saponification value determination?

Reagent required for Saponification reaction to occur
Acts as co-solvent (same concept as in acid value)

Why do we do a blank?
To compensate for loss of KOH due to reaction with atmospheric CO2 & formation
of KHCO3 /K2CO3

Requirements for a Saponification reaction:

Heat
Time (30 min)
Alcoholic KOH
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There are FIVE standard values to assess quality of oils. THREE of them are
discussed this lab.

1. Acid Value
2. Saponification Value
3. Ester Value
4. Iodine Value
5. Peroxide Value

23
 Ester value
Definition:
The ester value IE is the number of mg of potassium hydroxide required to
saponify the esters present in 1 g of the substance.

Determination of the ester value for virgin linseed oil sample

IE = IS-IA

For virgin linseed oil, the ester value is 183 to 190

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 General Report for Oil
Calculation

RULE = ………show substitution…….= …… (unitless)

e.g. Is = 28.05 (n2-n1)/m = 28.05 (24.7-17) / 1.01 = 213.85

Comment

The …………. value is below/within/above the range ( ……write range….)

Recommendation

Sample is accepted/ rejected.

N.B. NO Abbreviations!!
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 Report
Calculation

Acid Value:

Saponification value:

Ester Value:

Comment

Recommendation

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