INTERNATIONAL ISO
STANDARD 18321
First edition
2015-05-15
Essential oils — Determination of
peroxide value
Huiles essentielles — Détermination de l’indice de peroxyde
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ISO 18321:2015
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Reference number
ISO 18321:2015(E)
© ISO 2015
ISO 18321:2015(E)
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ISO 18321:2015
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ISO 18321:2015(E)
Contents Page
Foreword......................................................................................................................................................................................................................................... iv
1 Scope.................................................................................................................................................................................................................................. 1
2 Normative references....................................................................................................................................................................................... 1
3 Terms and definitions...................................................................................................................................................................................... 1
4 Principle......................................................................................................................................................................................................................... 1
5 Reagents......................................................................................................................................................................................................................... 1
6 Apparatus...................................................................................................................................................................................................................... 2
7 Sampling......................................................................................................................................................................................................................... 2
8 Preparation of test sample.......................................................................................................................................................................... 2
9 Procedure..................................................................................................................................................................................................................... 2
9.1 Test portion................................................................................................................................................................................................. 2
9.2 Determination.......................................................................................................................................................................................... 3
9.3 Automated potentiometric titration...................................................................................................................................... 3
10 Expression of results......................................................................................................................................................................................... 3
10.1 Calculation................................................................................................................................................................................................... 3
10.2 Conversion from mmol/l to meq/kg..................................................................................................................................... 3
11 Precision........................................................................................................................................................................................................................ 4
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11.1 Repeatability.............................................................................................................................................................................................. 4
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11.2 Reproducibility........................................................................................................................................................................................ 4
Bibliography................................................................................................................................................................................................................................. 5
ISO 18321:2015
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ISO 18321:2015(E)
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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see [Link]/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any
patent rights identified during the development of the document will be in the Introduction and/or on
the ISO list of patent declarations received (see [Link]/patents).
Any trade name used in this document is information given for the convenience of users and does not
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For an explanation on the meaning of ISO specific terms and expressions related to conformity
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to Trade (TBT), see the following URL: Foreword — Supplementary information.
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The committee responsible for this document is ISO/TC 54, Essential oils.
ISO 18321:2015
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iv © ISO 2015 – All rights reserved
INTERNATIONAL STANDARD ISO 18321:2015(E)
Essential oils — Determination of peroxide value
1 Scope
This International Standard specifies a method for the determination of the peroxide value in an
essential oil. The peroxide value is a measure of the oxidation present.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
ISO 212, Essential oils — Sampling
ISO 356, Essential oils — Preparation of test samples
ISO 3696, Water for analytical laboratory use — Specification and test methods
3 Terms and definitions
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For the purpose of this document, the following terms and definitions apply.
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3.1
peroxide value ISO 18321:2015
Ip [Link]
number that expresses, in millimolesa443d0ca6995/iso-18321-2015
(or milliequivalents), the quantity of peroxide contained in 1 000 ml
of the substance
4 Principle
It is a redox titration of the iodometry type. The iodide ions added to the essential oil oxidize when
reacting with peroxides, obtaining iodine which is titrated with thiosulphate. It can be carried out by
volumetric or potientiometric titration.
Potientiometric titration is particularly recommended for highly coloured essential oils for which are
difficult to appreciate the end point of the coloured indicator (e.g. vetiver essential oil).
5 Reagents
During the analysis, only reagents of recognized analytical grade and reverse osmosis or distilled or
deionized water of Grade 3, as defined in ISO 3696, should be used.
5.1 Trichloromethane (chloroform), 99 % (volume/fraction), or cyclohexane, 99,5 %
(volume/fraction) for laboratories with restrictions on the use of chloroform.
5.2 Glacial acetic acid, 99,5 % (volume/fraction). Degassed in an ultrasonic bath or by purging with a
current pure and dry inert gas (carbon dioxide or nitrogen).
5.3 Potassium iodide, saturated solution of potassium iodide in deionised water, freshly prepared.
The solution must remain saturated (undissolved crystals must be present). The solution has to be kept
protected from light.
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ISO 18321:2015(E)
5.4 Sodium thiosulfate solution, 0,01 mol/l (0,01 N) or 0,1 mol/l (0,1 N).
5.5 Coloured standard indicator, starch solution 1 % (volume/fraction). It is not necessary for
potentiometric titration.
6 Apparatus
Usual laboratory apparatus and, in particular, the following.
6.1 Balance, ±1 mg.
6.2 Erlenmeyer flask, of capacity 250 ml.
6.3 Shaker.
6.4 Pipettes, of capacity 1 ml, 10 ml, graduated in 0,1 ml.
6.5 Burette, of capacity 10 ml graduated in 0,05 ml.
6.6 Test tubes, of capacity 50 ml, 100 ml.
6.7 Potentiometer. iTeh STANDARD PREVIEW
7 Sampling
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Sampling is not included in the method specified ISO 18321:2015
in this International Standard. A recommended
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sampling method is given in ISO 212.
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It is important that the laboratory receives a representative sample, not damaged or modified during
the transport or storage, before the arrival at the laboratory.
8 Preparation of test sample
The test sample shall be prepared according to ISO 356.
The test sample for the determination of peroxide value shall be taken first and the peroxide value shall
be determined immediately. Homogenize the sample, preferably without heating and without aireation.
Avoid direct solar radiation. Heat solid samples carefully to 10 °C above their melting point. Samples
with visible impurities shall be filtered.
For some products, the amount of extracted essential oil can be lower than 5 g, or the peroxide value of
the essential oil can be over 30 meq of active oxygen per kilogram. In these cases, the user should choose
a smaller sample mass.
9 Procedure
9.1 Test portion
Introduce in an Erlenmeyer flask 10 ml of essential oil to be titrated (see 6.1, 6.3).
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ISO 18321:2015(E)
9.2 Determination
Add with a test tube (see 6.6) 20 ml of trichloromethane or cyclohexane, 30 ml of glacial acetic acid
and 1 ml of potassium iodide, saturated solution (see 5.1, 5.2, 5.3) and add 2 drops starch solution if
volumetric titration is carried out.
Shake for approximately 1 min. The dissolution acquires an orangey colour.
Add about 100 ml of distilled water.
Tritate with sodium thiosulfate solution. Use 0,1 mol/l when the expected peroxide value are over 20 mmol/l.
Use 0,01 mol/l sodium thiosulfate solution when the expected peroxide value are less than 20 mmol/l.
The end point is obtained when the dissolution becomes white.
Carry out a blank titration under the same conditions. No more than 0,5 ml of 0,01 mol/l sodium
thiosulfate solution should be consumed for this purpose.
9.3 Automated potentiometric titration
Many laboratories often use automated equipment for titration. In this case, some points have to be
taken into consideration.
— If the system works with autosampler, it is strictly necessary to use amber beaker glasses in order
to stop the formation of Iodine while the sample remains in the tray.
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— If the addition of potassium iodide wants to be automated, prepare 70 % (volume/fraction) solution
in distilled water instead of saturated solution, fill an opaque bottle, and replace it every week.
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Therefore, the addition of potassium iodide in 9.2 shall be 10 ml instead of 1 ml.
ISO 18321:2015
10 Expression of results
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10.1 Calculation
The peroxide value, Ip, in mmol/l, is given by Formula (1):
Ip = ( V1 − V0 ) × ( N titrant ) × 50 (1)
where
V1 is the volume, in millilitres, of titrant sodium thiosulphate, used in the main test;
V0 is the volume, in millilitres, of titrant sodium thiosulphate, used in the blank test;
Ntitrant is the concentration of titrant, sodium thiosulphate, 0,1 N (see 5.4).
10.2 Conversion from mmol/l to meq/kg
The conversion from mmol/l to meq/kg is calculated by Formula (2):
meq/kg = mmol/l × 2/density (kg/l) (2)
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11 Precision
11.1 Repeatability
The absolute difference between two independent single test results, obtained using this method on the
same essential oil tested in the same laboratory by the same operator using the same equipment within
a short period of time, was not, in more than 5 % in absolute values or 0,1 in relative values
11.2 Reproducibility
The absolute difference between two single test results, obtained using the same method on the same
essential oil tested in different laboratories with different operators using different equipment, was not,
in more than 10 % in absolute values or 0,3 in relative values.
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Bibliography
[1] ISO 385-11), Laboratory glassware — Burettes — Part 1: General requirements
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1) This standard has been replaced by ISO 385.
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