INTERNATIONAL ISO

STANDARD 709

Second edition
2001-09-01

Essential oils — Determination of ester

value
Huiles essentielles — Détermination de l'indice d'ester

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ISO 709:2001
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Reference number
ISO 709:2001(E)

© ISO 2001
ISO 709:2001(E)

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Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO
member bodies). The work of preparing International Standards is normally carried out through ISO technical
committees. Each member body interested in a subject for which a technical committee has been established has
the right to be represented on that committee. International organizations, governmental and non-governmental, in
liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical
Commission (IEC) on all matters of electrotechnical standardization.

International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 3.

Draft International Standards adopted by the technical committees are circulated to the member bodies for voting.
Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote.

Attention is drawn to the possibility that some of the elements of this International Standard may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights.

International Standard ISO 709 was prepared by Technical Committee ISO/TC 54, Essential oils.

This second edition cancels and replaces the first edition (ISO 709:1980), which has been technically revised.
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Annex A of this International Standard is for information only.
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ISO 709:2001
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INTERNATIONAL STANDARD ISO 709:2001(E)

Essential oils — Determination of ester value

1 Scope
This International Standard specifies a method for the determination of the ester value of an essential oil.

This method is not applicable to essential oils containing lactones or an appreciable proportion of aldehydes.

2 Normative references
The following normative documents contain provisions which, through reference in this text, constitute provisions of
this International Standard. For dated references, subsequent amendments to, or revisions of, any of these
publications do not apply. However, parties to agreements based on this International Standard are encouraged to
investigate the possibility of applying the most recent editions of the normative documents indicated below. For
undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC
maintain registers of currently valid International Standards.
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ISO 356, Essential oils — Preparation of test samples
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ISO 385-1, Laboratory glassware — Burettes — Part 1: General requirements
ISO 709:2001
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ISO 1242, Essential oils — Determination of acid value
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3 Term and definition

For the purposes of this International Standard, the following term and definition applies.

3.1
ester value
EV
number of milligrams of potassium hydroxide required to neutralize the acids liberated by the hydrolysis of esters
present in 1 g of the essential oil

4 Principle
The esters present in the essential oil are hydrolysed by heating under specified conditions with an excess of a
standard volumetric ethanolic potassium hydroxide solution. The excess alkali is determined by back titration with a
standard solution of hydrochloric acid.

5 Reagents
During the analysis, use only reagents of recognized analytical grade and distilled or demineralized water or water
of equivalent purity.

5.1 Ethanol, 95 % (volume fraction) at 20 °C, freshly neutralized with the potassium hydroxide solution (5.2), in
the presence of the coloured indicator (5.4) used for the determination.

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ISO 709:2001(E)

5.2 Potassium hydroxide, standard volumetric ethanolic solution, c(KOH) = 0,5 mol/l at 20 °C, freshly
restandardized before each series of tests.

5.3 Hydrochloric acid, standard volumetric solution, c(HCI) = 0,5 mol/l at 20 °C.

It is important that the reagent be taken at the specified temperature of 20 °C, particularly the ethanolic solution of
potassium hydroxide, as the volume varies greatly with temperature.

5.4 Coloured indicator.

Use phenolphthalein, 2 g/l solution in ethanol (5.1), or phenol red, 0,4 g/l solution in ethanol, 20 % (volume
fraction), if the essential oil has components that contain phenol groups.

NOTE This particular case will be specified in the specific standard for the essential oil concerned.

6 Apparatus
Usual laboratory apparatus and, in particular, the following.

6.1 Saponification flask, with ground glass neck, of alkali-resistant glass, of capacity 100 ml to 250 ml, to which
can be fitted a ground glass air condenser at least 1 m in length with 1 cm to 1,5 cm internal diameter.

If necessary, and particularly for the essential oils with high light fractions and depending on the time placed in the
boiling water bath, the glass tube may be replaced by a water-cooled reflux condenser.
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6.2 Test tubes, of capacity 5 ml.
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6.3 Burettes, of capacity 25 ml, graduated in 0,05 ml, complying with the requirements of ISO 385-1, class B.
ISO 709:2001
6.4 Boiling water bath. [Link]
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6.5 Analytical balance, accurate to the nearest 0,001 g.

6.6 Potentiometer.

7 Sampling
Sampling is not included in the method specified in this International Standard. A recommended sampling method
is given in ISO 2121).

It is important that the laboratory receive a representative sample, not damaged or modified during transport or
storage before the arrival at the laboratory.

8 Preparation of test sample

The test sample shall be prepared according to ISO 356.

1) ISO 212, Essential oils — Sampling.

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 ISO 709:2001(E)

9 Procedure

9.1 Test portion

Weigh, to the nearest 0,005 g, 2 g of the test sample.

The test portion may be different from this, if so specified in the specific standard for the essential oil concerned.

9.2 Blank test

Carry out a blank test, in parallel with the determination (9.3), under the same conditions and using the same
reagents. (See 9.3.3.)

9.3 Determination

9.3.1 Introduce the test portion (9.1) into the saponification flask (6.1). Add from the burette (6.3) 25 ml of the
potassium hydroxide solution (5.2) (see note) and fragments of pumice stone or porcelain.

NOTE If the test portion has been retained from the determination of the acid value, it will not be necessary to neutralize it
before adding the potassium hydroxide.

For oils with a high ester value, increase the volume of the potassium hydroxide solution (5.2) used so that
(V0 – V1) (see clause 10) is at least equal to 10 ml.

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For oils with a low ester value, increase the mass of the test portion used.

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Attach the air condenser or water-cooled reflux condenser, and place the flask in the boiling water bath (6.4) for a
time depending on the essential oil analysed. This time is mentioned in the specification for the oil to be tested.
ISO 709:2001
Allow to cool and remove the tube. Add 20 ml of water and 5 drops of the phenolphthalein solution, or of the phenol
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red solution (5.4) if the essential oil contains phenols or compounds with phenolic groups.
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9.3.2 Titrate the excess potassium hydroxide with the hydrochloric acid (5.3).

9.3.3 This determination may be carried out with the solution resulting from the determination of the acid value,
which can be used as the blank test, by adding 5 ml of ethanol (5.1) in this blank test before the addition of the
25 ml of potassium hydroxide solution (this volume corresponds to the volume introduced during the determination
of the acid value).

9.4 Potentiometry

Potentiometry may be used for all the essential oils, but it is particularly recommended for highly coloured essential
oils for which it is difficult to appreciate the endpoint of the coloured indicator (e.g. vetiver oil). In this case, the
same reagents and apparatus shall be used.

NOTE These special cases will be established in the specific standards for the essential oils concerned.

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ISO 709:2001(E)

10 Expression of results

10.1 Calculation

10.1.1 Ester value

The ester value (EV) is given by the formula

28,05
EV =
m
V 0 – V1  – AV

where

V0 is the volume, in millilitres, of hydrochloric acid (5.3) used for the blank test (9.2);

V1 is the volume, in millilitres, of hydrochloric acid (5.3) used for the determination (9.3.2);

m is the mass, in grams, of the test portion;

AV is the acid value determined according to ISO 1242.

The mass fraction of ester, w, as a percentage, with respect to a stated ester, is given by the formula

M r × EV
w=
561
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where
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ISO 709:2001
Mr is the relative molecular mass of the ester used to express the results conventionally;
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EV is the ester value calculated as above.

Express the ester value to two significant figures when it is less than 100, and to three significant figures when it is
100 or more.

10.1.2 Ester value determined after the acid value

When the determination is carried out on the solution resulting from the determination of the acid value, the ester
value (EV) is obtained by the formula

28,05
EV = (V 0 - V1 ¢ )
m

where V1' is the volume, in millilitres, of hydrochloric acid (5.3) used in the new determination.

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 ISO 709:2001(E)

11 Test report

The test report shall state:

¾ all information necessary for the complete identification of the sample;

¾ the sampling method used, if known;

¾ the test method used, with reference to this International Standard;

¾ all operating details not specified in this International Standard, or regarded as optional, together with details of
any incidents which may have influenced the test result(s);

¾ the test result(s) obtained.

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