EN ISO 29822:2014

SLOVENSKI STANDARD
01-september-2014
5DVWOLQVNHPDãþREHLQROMD,]RPHULþQLGLDFLOJOLFHUROL'RORþHYDQMHUHODWLYQLK
GHOHåHYLQGLDFLOJOLFHURORY,62
Vegetable fats and oils - Isomeric diacylglycerols - Determination of relative amounts of
1,2- and 1,3-diacylglycerols (ISO 29822:2009)
Tierische und pflanzliche Fette und Öle - Isomere Diacylglyceride - Bestimmung der
relativen Menge von 1,2- und 1,3- Diacylglyceride (ISO 29822:2009)
Corps gras d'origine végétale - Diacylglycérols isomériques - Détermination des teneurs
relatives en 1,2- et 1,3-diacylglycérols (ISO 29822:2009)
Ta slovenski standard je istoveten z: EN ISO 29822:2014
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 29822
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2014
ICS 67.200.10
English Version
Vegetable fats and oils - Isomeric diacylglycerols - Determination
of relative amounts of 1,2- and 1,3-diacylglycerols (ISO
29822:2009)
Corps gras d'origine végétale - Diacylglycérols isomériques Tierische und pflanzliche Fette und Öle - Isomere
- Détermination des teneurs relatives en 1,2- et 1,3- Diacylglyceride - Bestimmung der relativen Menge von 1,2-
diacylglycérols (ISO 29822:2009) und 1,3- Diacylglyceride (ISO 29822:2009)
This European Standard was approved by CEN on 11 April 2014.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 29822:2014 E
worldwide for CEN national Members.

Contents Page
Foreword .3
Foreword
The text of ISO 29822:2009 has been prepared by Technical Committee ISO/TC 34 “Food products” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 29822:2014 by
Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products -
Methods of sampling and analysis” the secretariat of which is held by AFNOR.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by October 2014, and conflicting national standards shall be withdrawn at
the latest by October 2014.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 29822:2009 has been approved by CEN as EN ISO 29822:2014 without any modification.

INTERNATIONAL ISO
STANDARD 29822
First edition
2009-03-15
Vegetable fats and oils —
Isomeric diacylglycerols —
Determination of relative amounts
of 1,2- and 1,3-diacylglycerols
Corps gras d'origine végétale — Diacylglycérols isomériques —
Détermination des teneurs relatives en 1,2- et 1,3-diacylglycérols

Reference number
ISO 29822:2009(E)
©
ISO 2009
ISO 29822:2009(E)
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ii © ISO 2009 – All rights reserved

ISO 29822:2009(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 29822 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
INTERNATIONAL STANDARD ISO 29822:2009(E)

Vegetable fats and oils — Isomeric diacylglycerols —
Determination of relative amounts of 1,2- and 1,3-
diacylglycerols
1 Scope
This International Standard specifies the determination of the degree of isomerization of diacylglycerols in
vegetable fats and oils. 1,2-Diacylglycerols are transformed to the more stable 1,3-isomers during storage or
due to acidic catalysed reaction.
The mass fraction of 1,2-diacylglycerols can be used as a quality criterion for vegetable fats and oils.
2 Normative references
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 661, Animal and vegetable fats and oils — Preparation of test sample
3 Terms and definitions
For the purposes of this International Standard, the following terms and definitions apply.
3.1
degree of isomerization
mass fraction of the peak areas of all 1,2-diacylglycerols (C , C , C ) relative to the sum of the peaks of all
32 34 36
diacylglycerols (C , C , C ).
32 34 36
NOTE The mass fraction is expressed as a percentage to one decimal place.
4 Principle
A miniaturized column chromatography on a silica gel column is used to separate the isomeric diacylglycerols
as the more polar fraction from the major part of other lipids. The peak areas of 1,2- and 1,3-isomers are
determined by gas chromatography after silylation. Only C -, C - and C -diacylglycerols are taken into
32 34 36
account.
5 Reagents
WARNING — Attention is drawn to the regulations which specify the handling of hazardous
substances. Technical, organizational and personal safety measures shall be followed.
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and distilled or
demineralized water or water of equivalent purity.
ISO 29822:2009(E)
1)
5.1 Silica gel 60 , for column chromatography, (0,063 to 0,100 mm).
5.1.1 Silica gel 60, with a moisture content, w = 5 % mass fraction.
H O
Activate the silica gel by heating overnight at (160 ± 5) °C. After heating, place the silica gel in a desiccator for
cooling and then transfer the silica gel to a stoppered flask. Add 5 % mass fraction of water and shake until no
lumps can be seen and the powder flows freely (1 h in an automatic shaking machine). Store the conditioned
silica gel overnight before use.
5.2 Cotton wool, defatted.
2)
5.3 Reference substances .
5.3.1 Dilaurin (1,3-dilauroyl-sn-glycerol).
5.3.2 Dipalmitin (1,2- and 1,3-dipalmitoyl-sn-glycerol, mixed isomers, w ≈ 99 % mass fraction).
5.3.3 Distearin (1,2- and 1,3-distearoyl-sn-glycerol, mixed isomers, w ≈ 99 % mass fraction).
5.3.4 Diolein (1,2- and 1,3-dioleoyl-sn-glycerol, mixed isomers, w ≈ 99 % mass fraction).
5.4 Reference solutions:
For the reference substances dipalmitin and distearin, prepare a solution in toluene (mass concentration,
ρ = 1 mg/ml).
5.5 Isooctane (2,2,4-trimethylpentane).
5.6 Diethyl ether, eluent.
5.7 Diisopropyl ether.
5.8 Solvent mixture: isooctane/diisopropyl ether [volume fraction ϕ (isooctane) = 85 ml/100 ml and
ϕ (diisopropyl ether) = 15 ml/100 ml].
5.9 1-Methylimidazole.
5.10 N-Methyl-N-(trimethyl-silyl)heptafluorobutyramide (MSHFBA).
5.11 Silylating reagent: 50 µl 1-methylimidazole in 1 ml of MSHFBA.
5.12 Toluene.
6 Apparatus
Usual laboratory equipment and, in particular, the following.
6.1 Analytical balance, capable of weighing to the nearest 0,001 g and displaying 0,000 1 g.

1) Merck Art.-No 612-1484 or 613-2623 (www.merck.de) 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. Equivalent products may be used if they can be shown to lead to the same results.
2) A suitable supplier is Sigma-Aldrich (www.sigmaaldrich.com). This information is given for the convenience of users
of this International Standard and does not constitute an endorsement by ISO of this supplier.
2 © ISO 2009 – All rights reserved

ISO 29822:2009(E)
6.2 Desiccator, for storing adsorbents after heating.
3)
6.3 Pipette tip , capacity 5 ml, length approx. 150 mm.
6.4 Pointed flask, capacity 10 ml, with ground-glass neck and rounded bottom.
6.5 Beaker, capacity 10 ml.
6.6 Rotary evaporator.
6.7 Gas chromatograph for capillary columns, split injector, flame ionization detector (FID) and suitable
integration system.
Do not use on-column or splitless injectors, as this results in isomerization of 1,2-diacylglycerols.
6.8 Fused silica capillary column, for gas chromatography (internal diameter, 0,25 mm or 0,32 mm,
length 12 m, 15 m, 30 m or 60 m) coated with 5 % diphenyl-, 95 % dimethylpolysiloxane, 0,1 µm film thickness.
Other columns of similar polarity and selectivity may also be used.
7 Sample
7.1 Sampling
Sampling is not part of the method specified in this International Standard. A recommended sampling method
[1]
is given in ISO 5555 .
It is important that the laboratory receive a sample which is truly representative and has not been damaged or
changed during transport or storage.
7.2 Preparation of test sample
Prepare the test sample in accordance with ISO 661.
8 Procedure
8.1 Preparation of the silica gel chromatography column
Insert a small stopper of cotton wool (5.2) approx. 5 mm high, in the lower part of a pipette tip (6.3) followed by
1 g silica gel 60 (5.1). Cover the silica layer with a 5 mm high stopper of cotton wool. Compress the filling by
slight pressing with a flat ended rod.
8.2 Separation of the fraction containing non polar lipids
8.2.1 Weigh, to the nearest 0,1 mg, about 100,0 mg of the test sample into a beak
...