ISO 29822-2:2026

International
Standard
ISO 29822-2
First edition
Vegetable fats and oils —
2026-03
Determination of the content
and relative amounts of 1,2- and
1,3-diacylglycerols —
Part 2:
Isolation by solid phase extraction
(SPE)
Reference number
© ISO 2026
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ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Reagents . 2
6 Apparatus . 3
7 Sample . 3
7.1 Sampling .3
7.2 Preparation of test sample .3
8 Procedure . 4
8.1 Separation of the fraction containing non-polar lipids .4
8.2 Preparation of trimethylsilyl ethers (silylation) .4
8.3 Gas chromatography .4
8.4 Identification of diacylglycerol isomers .5
8.5 Initial .5
8.6 Determination of the peak areas of 1,2- and 1,3- diacylglycerols in the oil.5
9 Precision of the method . 5
9.1 Interlaboratory testing .5
9.2 Repeatability .6
9.3 Reproducibility .6
10 Test report . 6
Annex A (informative) Example of a typical chromatogram . 7
Annex B (informative) Results of an interlaboratory study . 8
Bibliography .10

iii
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
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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 document should be noted. This document was drafted in accordance with the editorial rules of the
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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
A list of all parts in the ISO 29822 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
Introduction
The ISO 29822 series can be used for the determination of the content and relative amounts of 1,2- and 1,3-
diacylglycerols.
ISO 29822 uses a miniaturized column chromatography on a silica gel column 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 C32-, C34- and C36-diacylglycerols
are taken into account.
This document separates the diacylglycerol fraction by means of a solid phase extraction (SPE) column with
diol phase, followed by analysis on capillary gas chromatography after silylation. Only the peak areas of 1,2-
and 1,3- isomers of C34- and C36-diacylglycerols are taken into account. Less solvent and preparation are
needed than with silica gel column separation where careful preparation with the appropriate percentage of
water is necessary to avoid isomerization.

v
International Standard ISO 29822-2:2026(en)
Vegetable fats and oils — Determination of the content and
relative amounts of 1,2- and 1,3-diacylglycerols —
Part 2:
Isolation by solid phase extraction (SPE)
1 Scope
This document specifies the determination of the absolute value of diacylglycerols as well as 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.
NOTE Results obtained using this document have not been compared with results obtained using ISO 29822.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements 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 document, the following terms and definitions apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1
degree of isomerization
mass fraction of the peak areas of all 1,2-diacylglycerols (C34, C36) relative to the sum of the peaks of all
diacylglycerols (C34, C36)
Note 1 to entry: The mass fraction is expressed as a percentage to one decimal place.
Note 2 to entry: For oils with high content of palmitic acid, 1,2-diacylglycerols needs to include C32-diacylglycerols.
3.2
absolute content
mass fraction of peak areas of all diacylglycerols (C34, C36) relative to the sum of the peaks of all
diacylglycerols (C34, C36)
Note 1 to entry: The mass fraction is expressed in mg/kg to one decimal place.
Note 2 to entry: For oils with high content of palmitic acid, 1,2-diacylglycerols needs to include C32-diacylglycerols.

4 Principle
The diacylglycerol fraction is isolated by means of a solid phase extraction column with diol phase,
followed by analysis on capillary gas chromatography after silylation. Only the peak areas of 1,2- and 1,3-
isomers of C34- and C36-diacylglycerols are taken into account. For oils with high content of palmitic acid,
1,2-diacylglycerols should include C32-diacylglycerols.
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.
1)
5.1 Reference substances.
5.1.1 Dipalmitin (1,3-dipalmitoyl-sn-glycerol).
5.1.2 Dilaurin (1,3-dilauroyl-sn-glycerol). Internal standard used for oils with high content of palmitic
acid.
5.1.3 Dipalmitin (1,2- and 1,3-dipalmitoyl-sn-glycerol, mixed isomers, w ≈ 99 % mass fraction).
5.1.4 Distearin (1,2- and 1,3-distearoyl-sn-glycerol, mixed isomers, w ≈ 99 % mass fraction).
5.1.5 Diolein (1,2- and 1,3-dioleoyl-sn-glycerol, mixed isomers, w ≈ 99 % mass fraction).
5.2 Internal standard solutions.
To facilitate the dissolution of the internal standards, use an ultrasound bath.
5.2.1 Dipalmitin (1,3-dipalmitoyl-sn-glycerol) in diethyl ether (mass concentration, ρ = 0,1 mg/ml).
5.2.2 Dilaurin (1,3-dilauroyl-sn-glycerol) in diethyl ether (mass concentration, ρ = 0,1 mg/ml).
5.2.3 Dinonadecanoin (1,3-Dinonadecanoyl-rac-glycerol) in diethyl ether (mass
...