EN ISO 11701:2009
(Main)Vegetable fats and oils - Determination of phospholipids content in lecithins by HPLC using a light-scattering detector (ISO 11701:2009)
Vegetable fats and oils - Determination of phospholipids content in lecithins by HPLC using a light-scattering detector (ISO 11701:2009)
ISO 11701:2009 specifies a method for the quantitative determination of phospholipids content by high performance liquid chromatography (HPLC) using a diol column and a light-scattering detector.
The method is applicable to crude, oil-containing lecithins, and to oil-free lecithins and lecithin fractions from vegetable fats and oils.
The method is not applicable to animal and ruminant lecithins and enzymatically hydrolysed lecithins as the peak separation of lysophosphatidylethanolamine (LPE), lysophosphatidylinositol (LPI) and lysophosphatidic acid (LPA) is insufficient.
Pfanzliche Fette und Öle - Bestimmung des Gehaltes an Phospholipiden in Lecithinen durch HPLC mittels eines Lichtstreudetektors (ISO 11701:2009)
Diese Internationale Norm legt ein Verfahren zur quantitativen Bestimmung des Gehaltes an Phospholipiden durch Hochleistungsflüssigkeitschromatographie (HPLC) unter Anwendung einer Diol-Säule und eines Licht¬streudetektors fest.
Das Verfahren ist auf rohe ölhaltige Lecithine, ölfreie Lecithine und Lecithin-Fraktionen aus pflanzlichen Fetten und Ölen anwendbar.
Das Verfahren ist nicht anwendbar auf tierische Lecithine, Lecithine in Futtermitteln für Wiederkäuer und enzymatisch hydrolysierte Lecithine, da die Peaktrennung von Lysophosphatidylethanolamin (LPE), Lyso-phosphatidylinositol (LPI) und Lysophosphatidsäure (LPA) unzureichend ist.
Corps gras d'origine végétale - Détermination de la teneur en phospholipides dans les lécithines par CLHP avec détecteur à diffusion de la lumière (ISO 11701:2009)
L'ISO 11701:2009 spécifie une méthode de dosage quantitatif des phospholipides par chromatographie liquide à haute performance (CLHP) en utilisant une colonne de diol et un détecteur à diffusion de la lumière.
La méthode est applicable aux lécithines brutes contenant de l'huile, aux lécithines exemptes d'huile et aux fractions de lécithine provenant de corps gras d'origine végétale.
La méthode n'est applicable ni aux lécithines d'origine animale et provenant de ruminants, ni aux lécithines hydrolysées par des enzymes, la séparation des pics de la lysophosphatidyléthanolamine (LPE), du lysophosphatidylinositol (LPI) et de l'acide lysophosphatidique (LPA) étant insuffisante.
Rastlinske in živalske maščobe in olja - Določevanje fosfolipidov v lecitinih s tekočinsko kromatografijo visoke ločljivosti (HPLC) in z uporabo detektorja razpršitve svetlobe (ISO 11701:2009)
Ta mednarodni standard določa metodo za kvantitativno določanje vsebnosti fosfolipidov v lecitinih s tekočinsko kromatografijo visoke ločljivosti (HPLC), ki uporablja diol kolono in detektor razpršitve svetlobe. Ta metoda velja za surove lecitine, ki vsebujejo olje, in lecitine, ki ne vsebujejo olja ter frakcije lecitinov iz rastlinskih maščob in olj. Ta metode na velja za živalske in prežvekovalske lecitine in encimatske hidrolizirane lecitine, ker ločljivost vrha lisofosfatdiletanolamina (LPE), lisofosfatdilinositola (LPI) in lisofosfatne kisline (LPA) ne zadostuje.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-marec-2010
5DVWOLQVNHLQåLYDOVNHPDãþREHLQROMD'RORþHYDQMHIRVIROLSLGRYYOHFLWLQLKV
WHNRþLQVNRNURPDWRJUDILMRYLVRNHORþOMLYRVWL +3/& LQ]XSRUDERGHWHNWRUMD
UD]SUãLWYHVYHWOREH ,62
Vegetable fats and oils - Determination of phospholipids in lecithins by high performance
liquid chromatography (HPLC) using a light scattering detector (ISO 11701:2009)
Tierische und pflanzliche Fette und Öle - Bestimmung von Phospholipiden in pflanzlichen
Lecithinen durch HPLC mittels eines Lichtstreudetektors (ISO 11701:2009)
Corps gras d'origine végétale - Détermination des phospholipides dans les lécithines par
chromatographie liquide à haute performance (CLHP) utilisant un détecteur à diffusion
de la lumière (ISO 11701:2009)
Ta slovenski standard je istoveten z: EN ISO 11701:2009
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 11701
NORME EUROPÉENNE
EUROPÄISCHE NORM
December 2009
ICS 67.200.10
English Version
Vegetable fats and oils - Determination of phospholipids content
in lecithins by HPLC using a light-scattering detector (ISO
11701:2009)
Corps gras d'origine végétale - Détermination de la teneur Pfanzliche Fette und Öle - Bestimmung des Gehaltes an
en phospholipides dans les lécithines par CLHP avec Phospholipiden in Lecithinen durch HPLC mittels eines
détecteur à diffusion de la lumière (ISO 11701:2009) Lichtstreudetektors (ISO 11701:2009)
This European Standard was approved by CEN on 18 November 2009.
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: Avenue Marnix 17, B-1000 Brussels
© 2009 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11701:2009: E
worldwide for CEN national Members.
Contents Page
Foreword .3
Foreword
This document (EN ISO 11701:2009) has been prepared by Technical Committee ISO/TC 34 "Food products"
in collaboration with 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 June 2010, and conflicting national standards shall be withdrawn at
the latest by June 2010.
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 the United Kingdom.
Endorsement notice
The text of ISO 11701:2009 has been approved by CEN as a EN ISO 11701:2009 without any modification.
INTERNATIONAL ISO
STANDARD 11701
First edition
2009-12-15
Vegetable fats and oils — Determination
of phospholipids content in lecithins by
HPLC using a light-scattering detector
Corps gras d'origine végétale — Détermination de la teneur en
phospholipides dans les lécithines par CLHP avec détecteur à diffusion
de la lumière
Reference number
ISO 11701:2009(E)
©
ISO 2009
ISO 11701:2009(E)
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ii © ISO 2009 – All rights reserved
ISO 11701:2009(E)
Contents Page
Foreword .iv
1 Scope.1
2 Normative references.1
3 Terms and definitions .1
4 Principle .1
5 Reagents .2
6 Apparatus.2
7 Sampling .3
8 Preparation of test sample .3
9 Procedure.3
10 Calculation and expression of results .5
11 Precision of the method.5
12 Test report.6
Annex A (informative) HPLC chromatogram.7
Annex B (informative) Results of an interlaboratory test .8
Bibliography.11
ISO 11701:2009(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 11701 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
iv © ISO 2009 – All rights reserved
INTERNATIONAL STANDARD ISO 11701:2009(E)
Vegetable fats and oils — Determination of phospholipids
content in lecithins by HPLC using a light-scattering detector
1 Scope
This International Standard specifies a method for the quantitative determination of phospholipids content by
high performance liquid chromatography (HPLC) using a diol column and a light-scattering detector.
The method is applicable to crude, oil-containing lecithins, and to oil-free lecithins and lecithin fractions from
vegetable fats and oils.
The method is not applicable to animal and ruminant lecithins and enzymatically hydrolysed lecithins as the
peak separation of lysophosphatidylethanolamine (LPE), lysophosphatidylinositol (LPI) and lysophosphatidic
acid (LPA) is insufficient.
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 document, the following terms and definitions apply.
3.1
content of an individual phospholipid
mass fraction of N-acyl-phosphatidylethanolamine (N-acyl-PE) or phosphatidylcholine (PC) or
phosphatidylethanolamine (PE) or phosphatidylinositol (PI) or phosphatidic acid (PA) or
lysophosphatidylcholine (LPC), determined in accordance with the method specified in this International
Standard
NOTE The content is expressed in grams per 100 g, numerically equal to a percentage mass fraction.
4 Principle
The individual phospholipids are separated by HPLC using a diol column and a light-scattering detector. For
the purpose of quantification, a certified reference mixture is used.
ISO 11701:2009(E)
5 Reagents
WARNING — Comply with any local 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.
5.1 Water, HPLC grade.
5.2 n-Hexane, HPLC grade.
5.3 2-Propanol, HPLC grade.
5.4 Acetic acid, w , 99,8 % mass fraction.
min
5.5 Triethylamine.
5.6 Solvent mixture: A mixture of 80 ml n-hexane (5.2) and 20 ml 2-propanol (5.3) (volume fraction
ϕ = 80 ml/100 ml for n-hexane and ϕ = 20 ml/100 ml for 2-propanol) is used to dissolve the standards and
sample.
1)
5.7 Reference substance (external standard) ILPS-LE01 , mixed soy phospholipid reference standard,
is a certified reference mixture with defined contents of N-acyl-PE, PA, PE, PC, PI, and LPC.
5.8 Mobile phase for the HPLC.
5.8.1 Eluent A. Mix 814,2 ml of n-hexane (5.2), 170,0 ml of 2-propanol (5.3), 15 ml of acetic acid (5.4), and
0,8 ml of triethylamine (5.5) (volume fraction ϕ = 81,42 ml/100 ml for n-hexane, ϕ = 17,00 ml/100 ml for 2-
propanol, ϕ = 1,50 ml/100 ml for acetic acid, and ϕ = 0,08 ml/100 ml for triethylamine).
In order to obtain a reproducible eluent composition, it is recommended that the solvents be weighed out
taking into account their densities. For a batch size of 2,5 l: 1 341,4 g of n-hexane, 331,5 g of 2-propanol,
39,4 g of acetic acid, and 1,45 g (2,0 ml) of triethylamine.
5.8.2 Eluent B. Mix 844,2 ml of 2-propanol (5.3), 140 ml of water (5.1), 15,0 ml of acetic acid (5.4) and
0,8 ml of triethylamine (5.5) (volume fraction ϕ = 8
...
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