EN ISO 18395:2007
(Main)Animal and vegetable fats and oils - Determination of monoacylglycerols, diacylglycerols, triacylglycerols and glycerol by high-performance size-exclusion chromatography (HPSEC) (ISO 18395:2005)
Animal and vegetable fats and oils - Determination of monoacylglycerols, diacylglycerols, triacylglycerols and glycerol by high-performance size-exclusion chromatography (HPSEC) (ISO 18395:2005)
ISO 18395:2005 specifies a method for the determination of monoacylglycerols, diacylglycerols and triacylglycerols and also free glycerol by high-performance size-exclusion chromatography. It is applicable to products (e.g. emulsifiers) comprising monoacylglycerols and diacylglycerols as main constituents in concentrations greater than 10 %, and to triacylglycerols in a proportion of less than 20 %.
The method is not applicable to dairy fats or fats and oils having a wide range of fatty acid chain lengths, since diacylglycerols of short fatty acids have a lower molecular mass than monoacylglycerols of long-chain fatty acids.
The method has restricted applicability to acylglycerol mixtures based on caprylic and capric acids. Here, only the monoacylglycerol content and the free glycerol content can be determined.
Tierische und pflanzliche Fette und Öle - Bestimmung von Monoacylglycerinen, Diacylglycerinen, Triacylglycerinen und Glycerin durch Hochleistungs-Größenausschluss-Chromatographie (HPSEC) (ISO 18395:2005)
Corps gras d'origines animale et végétale - Détermination de la teneur en monoacylglycérides, en diacylglycérides, en triacylglycérides et en glycérol par chromatographie d'exclusion haute performance (CLHP d'exclusion) (ISO 18395:2005)
L'ISO 18395:2005 spécifie une méthode permettant de déterminer la teneur en monoacylglycérides, en diacylglycérides, en triacylglycérides et en glycérol libre par chromatographie liquide d'exclusion haute performance. Elle s'applique aux produits (par exemple les émulsifiants) renfermant comme constituants principaux des monoacylglycérides et des diacylglycérides à une concentration supérieure à 10 % ainsi que des triacylglycérides à une teneur inférieure à 20 %.
La méthode ne s'applique pas aux corps gras laitiers ou aux corps gras et aux huiles ayant une gamme étendue d'acides gras, car les diacylglycérides présents dans les acides gras à chaîne courte peuvent avoir une masse moléculaire inférieure à celle des monoacylglycérides présents dans les acides gras à chaîne longue.
L'application de la méthode est limitée à ce qui concerne les mélanges glycéridiques à base d'acides caprylique et caprique. Dans ceux-ci, seule la teneur en monoacylglycérides et en glycérol libre peut être déterminée.
Rastlinske in živalske maščobe in olja - Določevanje monoacilglicerolov, diacilglicerolov, triacilglicerolov in glicerola s kromatografijo visoke ločljivosti z ločevanjem po velikosti (HPSEC) (ISO 18395:2005)
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-november-2007
5DVWOLQVNHLQåLYDOVNHPDãþREHLQROMD'RORþHYDQMHPRQRDFLOJOLFHURORY
GLDFLOJOLFHURORYWULDFLOJOLFHURORYLQJOLFHURODVNURPDWRJUDILMRYLVRNHORþOMLYRVWL]
ORþHYDQMHPSRYHOLNRVWL +36(& ,62
Animal and vegetable fats and oils - Determination of monoacylglycerols, diacylglycerols,
triacylglycerols and glycerol by high-performance size-exclusion chromatography
(HPSEC) (ISO 18395:2005)
Tierische und pflanzliche Fette und Öle - Bestimmung von Monoacylglycerinen,
Diacylglycerinen, Triacylglycerinen und Glycerin durch Hochleistungs-Größenausschluss
-Chromatographie (HPSEC) (ISO 18395:2005)
Corps gras d'origines animale et végétale - Détermination de la teneur en
monoacylglycérides, en diacylglycérides, en triacylglycérides et en glycérol par
chromatographie d'exclusion haute performance (CLHP d'exclusion) (ISO 18395:2005)
Ta slovenski standard je istoveten z: EN ISO 18395:2007
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 18395
NORME EUROPÉENNE
EUROPÄISCHE NORM
August 2007
ICS 67.200.10
English Version
Animal and vegetable fats and oils - Determination of
monoacylglycerols, diacylglycerols, triacylglycerols and glycerol
by high-performance size-exclusion chromatography (HPSEC)
(ISO 18395:2005)
Corps gras d'origines animale et végétale - Détermination Tierische und pflanzliche Fette und Öle - Bestimmung von
de la teneur en monoacylglycérides, en diacylglycérides, en Monoacylglycerinen, Diacylglycerinen, Triacylglycerinen
triacylglycérides et en glycérol par chromatographie und Glycerin durch Hochleistungs-Größenausschluss-
d'exclusion haute performance (CLHP d'exclusion) (ISO Chromatographie (HPSEC) (ISO 18395:2005)
18395:2005)
This European Standard was approved by CEN on 26 July 2007.
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 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 Management Centre has the same status as the
official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2007 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 18395:2007: E
worldwide for CEN national Members.
Foreword
The text of ISO 18395:2005 has been prepared by Technical Committee ISO/TC 34 "Agricultural
food products” of the International Organization for Standardization (ISO) and has been taken
over as EN ISO 18395:2007 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 February 2008, and conflicting national
standards shall be withdrawn at the latest by February 2008.
"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, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United
Kingdom.
Endorsement notice
The text of ISO 18395:2005 has been approved by CEN as EN ISO 18395:2007 without any
modifications.
INTERNATIONAL ISO
STANDARD 18395
First edition
2005-04-15
Animal and vegetable fats and oils —
Determination of monoacylglycerols,
diacylglycerols, triacylglycerols and
glycerol by high-performance size-
exclusion chromatography (HPSEC)
Corps gras d'origines animale et végétale — Détermination de la teneur
en monoacylglycérides, en diacylglycérides, en triacylglycérides et en
glycérol par chromatographie liquide d'exclusion (CLHP d'exclusion)
Reference number
ISO 18395:2005(E)
©
ISO 2005
ISO 18395:2005(E)
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
© ISO 2005
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
ISO's member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2005 – All rights reserved
ISO 18395:2005(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 18395 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
INTERNATIONAL STANDARD ISO 18395:2005(E)
Animal and vegetable fats and oils — Determination of
monoacylglycerols, diacylglycerols, triacylglycerols and
glycerol by high-performance size-exclusion chromatography
(HPSEC)
1 Scope
This International Standard specifies a method for the determination of monoacylglycerols, diacylglycerols and
triacylglycerols and also free glycerol by high-performance size-exclusion chromatography. It is applicable to
products (e.g. emulsifiers) comprising monoacylglycerols and diacylglycerols as main constituents in
concentrations >10 %, and to triacylglycerols in a proportion of < 20 %.
The method is not applicable to dairy fats or fats and oils having a wide range of fatty acid chain lengths, since
diacylglycerols of short fatty acids have a lower molecular mass than monoacylglycerols of long-chain fatty
acids.
The method has restricted applicability to acylglycerol mixtures based on caprylic and capric acids. Here, only
the monoacylglycerol content and the free glycerol content can be determined.
NOTE References [1] to [4] give background information.
2 Normative references
The following referenced document is 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 document, the following terms and definitions apply.
3.1
contents of monoacylglycerols, diacylglycerols, triacylglycerols and glycerol
proportion of monoacylglycerols, diacylglycerols, triacylglycerols and glycerol determined under the conditions
of this International Standard
NOTE The contents are expressed as a mass fraction (grams per 100 g) or as a percentage of all peaks.
4 Principle
The sample is dissolved in tetrahydrofuran (THF). The solution obtained is analysed by gel permeation
chromatography (GPC) using THF as the mobile phase. The acylglycerols and glycerol are separated
according to their molecular size. Detection is by means of a refractive index detector.
ISO 18395:2005(E)
5 Reagents
WARNING — Attention is drawn to regulations concerning the handling of hazardous substances.
Observe all technical, organizational and personal protective measures.
Use only reagents of recognized analytical grade, unless otherwise specified.
5.1 Tetrahydrofuran (THF), stabilized with 250 µl/l of BHT.
5.2 Standard substances
5.2.1 Glycerol (w W 99,5 %).
1)
5.2.2 Monoacylglycerols, diacylglycerols, triacylglycerols .
Standards used should be monoacylglycerols, diacylglycerols and triacylglycerols having a fatty acid
distribution as similar as possible to those present in the sample. However, the acylglycerol composition does
not have to be reproduced exactly. For most samples, monopalmitate and/or monostearate and/or
monooleate, dipalmitate and/or distearate and/or dioleate, tripalmitate and/or tristearate and/or trioleate are
sufficient.
The response factors for glycerol, monoacylglycerols, diacylglycerols and triacylglycerols are generally the
same under the conditions indicated in this International Standard, so that in most cases the use of
quantitative reference solutions is unnecessary and percentages by area may be used. However, it is
necessary to determine a response factor for glycerol when the concentration in the sample is > 3 %.
No response factors at all are employed in the European Pharmacopoeia for acylglycerols. This shall be
indicated in the test report.
6 Apparatus
Usual laboratory equipment and, in particular, the following.
6.1 Analytical balance, with a readability of 0,1 mg.
6.2 Volumetric flask, of 10 ml capacity.
6.3 Pipette, of 5 ml capacity.
6.4 Ultrasonic bath.
6.5 HPLC/GPC pump.
6.6 Injector, equipped with a sample loop of 20 µl capacity.
6.7 Detector, differential refractometer (RI detector).
6.8 GPC column combination, with an effective molecular mass up to 4 000 Da (e.g. three columns of
2)
300 mm × 7,5 mm Plgel , 5 µm, 100 Å).
It is also possible to use other columns (or column combinations) provided that the separation of the
monoacylglycerols and diacylglycerols and also of the diacylglycerols and triacylglycerols is ensured. The use
of only two columns is also possible to achieve the required separation.
1) Available for example from Sigma-Aldrich (http://www.sigmaaldrich.com/).
2) Available for example from Polymer Laboratories (http://www.polymerlabs.com/).
This information is given for the convenience of users of this International Standard and does not constitute an
endorsement by ISO of these products.
2 © ISO 2005 – All rights reserved
ISO 18395:2005(E)
6.9 Column oven (column thermostat).
6.10 Data integration and evaluation system.
7 Sampling
A representative sample should have been sent to the laboratory. It should not have been damaged or
changed during transport or storage.
Sampling is not part of the method specified in this International Standard. A recommended sampling method
[5]
is given in ISO 5555 .
8 Preparation of test sample
Prepare the test sample in accordance with ISO 661.
Before taking the test portion from the sample, mix the sample thoroughly to ensure homogeneity. For the
same reason, melt solid samples completely for proper mixing.
9 Procedure
9.1 Preparation of the sample solution
Weigh, to the nearest 0,1 mg, about 100 mg of the sample into a 10 ml volumetric flask. The concentration is
reported in milligrams of sample per 10 ml. After diluting to the mark with THF, place the flask in the ultrasonic
bath for 5 min to 10 min.
The clear solution obtained is directly injected into the GPC system.
9.2 Preparation of reference solutions
Weigh, to the nearest 0,1 mg, amounts of glycerol, monoacylglycerols, diacylglycerols and triacylglycerols
above and below the expected concentration in the sample into 10 ml volumetric flasks. After diluting to the
mark with THF, place the flasks in the ultrasonic bath for 5 min to 10 min.
The clear soluti
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.