INTERNATIONAL ISO

STANDARD 659

Fourth edition
2009-07-01

Oilseeds — Determination of oil content

(Reference method)
Graines oléagineuses — Détermination de la teneur en huile (Méthode
de référence)

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ISO 659:2009
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e7df879be007/iso-659-2009

Reference number
ISO 659:2009(E)

© ISO 2009
ISO 659:2009(E)

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 ISO 659:2009(E)

Contents Page

Foreword............................................................................................................................................................ iv
1 Scope ..................................................................................................................................................... 1
2 Normative references ........................................................................................................................... 1
3 Terms and definitions........................................................................................................................... 1
4 Principle ................................................................................................................................................. 1
5 Reagent .................................................................................................................................................. 2
6 Apparatus .............................................................................................................................................. 2
7 Sampling ................................................................................................................................................ 3
8 Preparation of test sample................................................................................................................... 3
8.1 Reduction of laboratory sample.......................................................................................................... 3
8.2 Predrying ............................................................................................................................................... 3
8.3 Test sample ........................................................................................................................................... 4
9 Procedure .............................................................................................................................................. 5
9.1 General................................................................................................................................................... 5
9.2
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Test portion ........................................................................................................................................... 5
9.3 Determination........................................................................................................................................ 6
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9.4 “Oil content” of impurities................................................................................................................... 7
10 Expression of results ...........................................................................................................................
ISO 659:2009 7
10.1 Method of calculation ........................................................................................................................... 7
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11 e7df879be007/iso-659-2009
Precision .............................................................................................................................................. 10
11.1 Interlaboratory test programme ........................................................................................................ 10
11.2 Repeatability........................................................................................................................................ 10
11.3 Reproducibility.................................................................................................................................... 10
12 Test report ........................................................................................................................................... 10
Annex A (informative) Results of interlaboratory tests on the determination of oil content ................... 11
Bibliography ..................................................................................................................................................... 12

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ISO 659:2009(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.

International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.

The main task of technical committees is to prepare International Standards. 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 document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.

ISO 659 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 2, Oleaginous
seeds and fruits and oilseed meals.
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This fourth edition cancels and replaces the third edition1) (ISO 659:1998), which has been technically revised.
The main change is the inclusion of an additional subclause (Subclause 8.3.5) for the preparation of the test
(standards.iteh.ai)
sample in the case of sunflower seed. This different procedure for sunflower seed includes an extra step, viz
measurement of the moisture content after grinding the seed. This is necessary to correct for the loss of
moisture caused by the heating of the seed which occurs ISO 659:2009
during grinding due to the particular physical nature
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1) Users should note that the front cover and foreword of ISO 659:1998 indicate erroneously that it is the second edition,
whereas it is in fact the third.

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INTERNATIONAL STANDARD ISO 659:2009(E)

Oilseeds — Determination of oil content (Reference method)

1 Scope
This International Standard specifies a reference method for the determination of the hexane extract (or light
petroleum extract), called the “oil content”, of oilseeds used as industrial raw materials. The procedure for
sunflower seed is different from those for other seeds as it includes an additional moisture content
determination after the seed has been ground to prepare the test sample.

The method has been tested on rapeseed, soya beans and sunflower seed. This does not, however, preclude
its applicability to other commercial seeds.

If required, the pure seeds and the impurities (see 9.4) can be analysed separately. In the case of groundnuts
(see 10.1.6), the pure seeds, the total fines, the non-oleaginous impurities and the oleaginous impurities can
be analysed separately.

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Normative references
(standards.iteh.ai)
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
ISO 659:2009
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ISO 658, Oilseeds — Determination of e7df879be007/iso-659-2009
content of impurities

ISO 664, Oilseeds — Reduction of laboratory sample to test sample

ISO 665, Oilseeds — Determination of moisture and volatile matter content

3 Terms and definitions

For the purposes of this International Standard, the following terms and definitions apply.

3.1
hexane extract
“oil content”
all the substances extracted under the operating conditions specified in this International Standard, expressed
as a percentage by mass of the product as received, or on the cleaned seed

NOTE On request, it may be expressed relative to the dry matter.

4 Principle
The oil is extracted from a test portion, in a suitable apparatus, with hexane or light petroleum. The solvent is
removed from the extract and the extract weighed. Due to the varied nature of seeds, there are some
variations in the procedure for different seeds.

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ISO 659:2009(E)

5 Reagent
5.1 Technical hexane, n-hexane or light petroleum, essentially composed of hydrocarbons with six
carbon atoms, of which less than 5 % distils below 40 °C and more than 95 % distils between 40 °C and 60 °C
or between 50 °C and 70 °C, and which has a bromine value of less than 1. The residue on complete
evaporation shall not exceed 2 mg per 100 ml.

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

6.1 Analytical balance, with an accuracy of ± 0,001 g.

6.2 Mechanical mill, easy to clean, appropriate for the nature of the oilseeds and allowing the oilseeds to
be ground to a uniform particle size without appreciable heating or change in moisture, volatile-matter or oil
content.

NOTE The following mills have been found to give satisfactory results:

⎯ the Christy & Norris 8" laboratory mill 2), with perforated-plate or bar-type screens depending on seed type (e.g.
0,8 mm perforated plate, 3 mm and 6 mm bar screens; see 8.3.2, 8.3.3 and 8.3.4);

⎯ the Retsch ZM 200 ultra-centrifugal mill 2), with a 1 mm screen for sunflower seed or with other screens depending on
seed type.
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Other mills may be satisfactory, provided they conform to the criteria given above.
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6.3 Mechanical microgrinder, capable of producing a fineness of grind of the oilseeds of less than 160 µm,
with the exception of the “shell”, particles of which ISO may659:2009
reach 400 µm. Coffee grinders and similar cutting-
action mechanical grinders shall not be used as microgrinders.
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NOTE A ball mill comprising a stainless-steel tube of volume approximately 120 ml and 1 cm, 2 cm and 3 cm
diameter steel balls (weighing approximately 7 g, 30 g and 130 g), depending on the sample being ground, and utilizing
extremely rapid shaking, has been found to be suitable.

If the microgrinder and its closure are designed for use with solvents, a small amount (approximately 20 ml) of
solvent may be added to the cylinder in order to increase the speed of the extraction.

6.4 Extraction thimble (cellulose) and cotton wool, free from matter soluble in hexane or light petroleum.

6.5 Suitable extraction apparatus, fitted with a flask of capacity 200 ml to 250 ml.

Straight-through extractors, for example the Butt, Smalley or Bolton-Williams type, are preferred for safety
reasons and have been found to give a more efficient extraction.

The use of other extractors is conditional upon the results of a test on a standard material of known oil content
to confirm the suitability of the apparatus. If a Soxhlet syphoning-type extractor is used then, for safety
reasons, a 250 ml flask should be utilized and the volume of solvent used shall be at least 150 ml. This should
prevent rapid boiling on syphoning, which could otherwise cause loss of solvent and/or part of the test portion.

6.6 Pumice stone, in small particles, or other anti-bumping granules, previously dried in an oven at
(130 ± 2) °C and cooled in a desiccator.

2) The Christy & Norris 8" mill and the Retsch ZM 200 mill are examples of suitable products available commercially.
This information is given for the convenience of users of this International Standard and does not constitute an
endorsement by ISO of these products.

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 ISO 659:2009(E)

6.7 Apparatus for safely removing solvent from extraction thimble (for example, in a current of inert
gas).

WARNING — This process should be carried out in a fume hood.

6.8 Electric heating bath (sand bath, water bath, heating mantle, etc.) or hot-plate.

6.9 Electrically heated oven, with thermostatic control, capable of being maintained at (103 ± 2) °C. The
oven shall be capable of being used at either atmospheric or reduced pressure (see 9.3.3).

6.10 Desiccator, containing an efficient desiccant (e.g. silica gel, with orange indicator, or P2O5).

6.11 Electrically heated oven, capable of being maintained at (130 ± 2) °C (for cottonseed, see 8.3.6).

6.12 Metal dish, flat-bottomed, of diameter 100 mm and height approximately 40 mm.

7 Sampling
Sampling is not part of the method specified in this International Standard. A recommended sampling method
is given in ISO 542.

It is important that the laboratory receive a sample which is truly representative and has not been damaged or
changed during transport and storage.

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8 Preparation of test sample
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8.1 Reduction of laboratory sample ISO 659:2009
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Prepare the test sample in accordance with ISO 664. If large non-oleaginous foreign bodies have been
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separated before reduction of the laboratory sample, make allowance for this in the calculation (see 10.1.5).
Depending on the requirements of the contract or the instructions, take the sample as received or after
separation of the impurities.

8.2 Predrying

8.2.1 The moisture content of the test portion (see 9.2) shall be less than a mass fraction of 10 % when
extraction of the oil (see 9.3) is begun.

NOTE Failure to observe this requirement can result in an incorrect result and invalidate the analysis.

Use a rapid screening method to assess the moisture content of the laboratory sample (see 8.1). If the
moisture mass fraction is greater than 10 %, reduce it to less than 10 % by drying the test sample (see 8.1) in
a dish in an oven maintained at a temperature not greater than 80 °C. Store the partially dried material in an
air-tight jar. Determine the moisture contents of the partially dried test sample and the original sample in
accordance with the method given in ISO 665 and make allowance for this in the final calculation (see 10.1.8).

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ISO 659:2009(E)

8.3 Test sample

8.3.1 Preliminary remarks

It is essential that oil extractions are carried out within 30 min of grinding, especially if the free fatty acid
content of the extracted oil is to be determined.

Care shall be taken to clean all mills thoroughly before and after each sample has been ground. Any material
adhering to the mill shall be incorporated into the bulk of the ground material.

NOTE In the following text, “seed” or “kernels” implies both whole seeds or kernels and broken fragments of them.

8.3.2 Palm kernels

Shell is considered to be part of the impurities, even when attached to the kernels. Shell and dirt shall be
separated quantitatively from the kernels before analysis.

Shell is hard and thus difficult to mill. It is almost impossible to obtain a homogeneous sample if kernels and
impurities are milled together. If the oil content of the whole sample, as received, is required, analyse the
kernels and impurities (shell and dirt) separately and calculate the oil content (see 10.1.3).

Prepare the test sample by grinding, separately, 600 g of the well-mixed kernels and all of the shell and dirt
separated during the quantitative determination of impurities in accordance with the method given in ISO 658.

The following milling techniques have been found to be suitable:

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⎯ for pure kernels: a mechanical mill (6.2) with a 6 mm bar screen;
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⎯ for shell and dirt: grind for 10 min in a microgrinder (6.3) using a 3 cm steel ball.
ISO 659:2009
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8.3.3 Copra (and coconut)
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Freeze the entire sample before milling in the mechanical mill (6.2).

A mechanical mill with a 6 mm screen has been found to be satisfactory. The length of the particles shall be
approximately 2 mm but shall not be greater than 5 mm. Mix the particles carefully and carry out the
determination without delay.

Care shall be taken to avoid condensation of moisture onto the seeds during and after grinding.

8.3.4 Large- and medium-sized seeds (illipe nuts, shea nuts, groundnuts, soya, etc., but NOT
sunflower seed or cottonseed)

Grind the test sample in the mechanical mill (6.2) until the major dimension of the particles obtained is not
greater than 2 mm. Reject the first particles (about one-twentieth of the sample), collect the rest, mix carefully
and carry out the determination without delay.

The following milling techniques have been found to be satisfactory:

⎯ for soya: a mechanical mill with a 0,8 mm perforated plate or with a 1 mm screen;

⎯ for groundnuts: a mechanical mill with a 3 mm bar screen (with samples containing a mass fraction of
more than about 45 % of oil, care shall be taken to avoid, as far as possible, the formation of a pasty
mass);

⎯ other seeds: a mechanical mill with a 6 mm bar screen.

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 ISO 659:2009(E)

The grinding of very oily seeds can often be improved if the seeds are first frozen at −10 °C to −20 °C, but
great care shall be taken to avoid condensation of moisture onto the seeds during and after grinding.

8.3.5 Sunflower seed

Determine the moisture and volatile-matter content of the sunflower seed (as received) on the whole seed in
accordance with ISO 665, taking care to observe strictly the drying periods laid down in the method specified
in ISO 665. Record the result of this moisture and volatile-matter determination (U1).

Grind the test sample in a mechanical mill (6.2). During grinding, the mill shall be fed slowly with the sunflower
seeds to prevent a paste being formed. A mechanical mill with a 1 mm screen has been found to be suitable.
If a screen-type mill is not used, sieve through a 2 mm sieve and regrind the material which does not pass
through the sieve until the major dimension of the particles is not greater than 2 mm. Collect all the sievings,
mix carefully and, without delay, determine the moisture and volatile-matter content in accordance with
ISO 665, taking care to observe strictly the drying periods laid down in ISO 665.

Record the result of the moisture and volatile-matter determination on the ground seed (U2) and carry out the
oil extraction on the ground seed as specified in Clause 9.

8.3.6 Cottonseed

Weigh, to the nearest 1 mg, into a tared metal dish (6.12) about 15 g of the sample as received. Place the
dish and seeds in the oven (6.11), previously heated to 130 °C, and leave to dry for 2 h at (130 ± 2) °C. Then
remove the dish from the oven and allow to cool in air for about 30 min. Transfer the whole test portion to a
mill (6.2) and grind to break up the seeds and linters. Transfer the ground material to an extraction thimble
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(6.4) and proceed with the determination.

8.3.7
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Small seeds (linseed, rapeseed, etc.)

Take a representative portion of approximately ISO 659:2009

100 g from the sample as received and grind it in a mechanical
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mill (6.2) so that no whole seeds remain. Ensure that any fine material remaining in the mill is incorporated
into the bulk of the ground material ande7df879be007/iso-659-2009
that the whole is thoroughly mixed. Care shall be taken that moisture
is not lost from the ground test sample.

The grinding time and speed (if variable) shall be predetermined for a particular mill and seed. The grinding
shall not cause segregation of the “meat” and “husks” of the seed, the test sample shall not be oily and at
least a mass fraction of 95 % shall pass a 1 mm sieve.

If the oil content in the seed free from impurities is required, remove the impurities by the method given in
ISO 658 and prepare at least 30 g of “clean” seed (including broken seed), as it is impracticable to prepare
100 g of very small seed such as sesame seed.

9 Procedure

9.1 General

Carry out two single determinations in accordance with 9.2 and 9.3 and check that the requirement concerning
repeatability (see 11.2) is satisfied. If it is not, repeat the determination on two further test portions taken from
a freshly prepared test sample (see 8.3). If the differences still exceed the value stated in 11.2, take as the
result the arithmetic mean of the four determinations carried out, provided that the maximum difference
between the individual results does not exceed 1,50 % in absolute value.

9.2 Test portion

9.2.1 Weigh, to the nearest 1 mg, (10 ± 0,5) g of the ground test sample (see 8.3). For treatment of
impurities, see 9.4.

© ISO 2009 – All rights reserved 5