EN ISO 11701:2009

• Abstract:

  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.

• Status: Published
• Publication Date: 14-Dec-2009
• Withdrawal Date: 29-Jun-2010
• ICS: 67.200.10 - Animal and vegetable fats and oils
• Technical Committee: CEN/TC 307 - Oilseeds, vegetables and animal fats and oils and their by-products - Methods of sampling and analys
• Drafting Committee: CEN/TC 307 - Oilseeds, vegetables and animal fats and oils and their by-products - Methods of sampling and analys
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 15-Dec-2009
• Completion Date: 15-Dec-2009
• Ref Project: SIST EN ISO 11701:2010 - Vegetable fats and oils - Determination of phospholipids in lecithins by high performance liquid chromatography (HPLC) using a light scattering detector (ISO 11701:2009)

• Refers: ISO 661:2003 - Animal and vegetable fats and oils — Preparation of test sample
• Effective Date: 25-Aug-2026
• Consolidated By: ISO 11701:2009 - Vegetable fats and oils — Determination of phospholipids content in lecithins by HPLC using a light-scattering detector
• Effective Date: 12-Feb-2026

Overview

EN ISO 11701:2009 is an internationally recognized standard developed by CEN and ISO for the quantitative determination of phospholipid content in lecithins derived from vegetable fats and oils. The standard specifies a validated method that uses high performance liquid chromatography (HPLC) with a diol column and a light-scattering detector. This analytical approach ensures accurate identification and quantification of individual phospholipids, supporting quality control and compliance in the food, feed, and lecithin processing industries.

Keywords: vegetable lecithins, phospholipid determination, HPLC, light-scattering detector, vegetable oils, lecithin analysis, standard methods

Key Topics

• Scope and Applicability

  • Applies to crude, oil-containing lecithins, oil-free lecithins, and lecithin fractions from vegetable sources.
  • Not suitable for animal lecithins, ruminant lecithins, or enzymatically hydrolysed lecithins due to insufficient peak separation for certain lysophospholipids.

• Principle of Method

  • Phospholipids in lecithins are separated on a diol HPLC column.
  • Quantification is achieved using a calibrated light-scattering detector and certified reference mixtures.

• Test Sample Preparation

  • Importance of representative, homogenized samples.
  • Samples are dissolved in specific solvent mixtures prior to analysis.

• Precision and Repeatability

  • Method precision is defined through international interlaboratory tests.
  • Parameters for repeatability and reproducibility ensure reliability across laboratories.

Applications

• Quality Control in Lecithin Production

  • Ensures accurate phospholipid profiling in food-grade and technical-grade lecithins.
  • Helps manufacturers verify that lecithin products meet regulatory and customer specifications.

• Food and Feed Industry

  • Useful for verifying the composition of lecithins used as emulsifiers in processed foods.
  • Enables producers to standardize product functionality and performance.

• Research and Development

  • Supports technical research in the development of new lecithin fractions or applications.
  • Facilitates investigation of phospholipid composition for nutritional labeling or functional food innovation.

• Supply Chain Compliance

  • Assists in certifying the authenticity and purity of vegetable lecithin fractions supplied to various industries.
  • Meets international requirements for consistent product analysis and reporting.

Related Standards

• ISO 661: Animal and Vegetable Fats and Oils - Preparation of Test Sample

  • Provides guidance on test sample preparation, an essential pre-requisite for reliable HPLC analysis.

• ISO 5555: Animal and Vegetable Fats and Oils - Sampling

  • Establishes standardized procedures for sample collection, crucial for representative testing.

• ISO 5725 Series: Accuracy (Trueness and Precision) of Measurement Methods and Results

  • Defines methodology for assessing repeatability and reproducibility of analytical methods.

• ISO 1042: Laboratory Glassware - One-mark Volumetric Flasks

  • Specifies requirements for volumetric equipment used in sample and standard solution preparation.

Practical Value

Adhering to EN ISO 11701:2009 provides the following practical advantages:

• Assures Global Acceptance: Use of a harmonized, internationally recognized test provides credibility in global supply chains.
• Supports Regulatory Compliance: Provides a scientific basis for compliance with food safety and ingredients regulations.
• Enhances Product Consistency: Detailed phospholipid profiling helps maintain product quality and consistency, key for industrial applications.
• Reduces Analytical Uncertainty: Standardized methodology minimizes variability and sources of error in phospholipid determination.

By implementing EN ISO 11701:2009, laboratories and industry stakeholders ensure that the determination of phospholipids in vegetable lecithins is accurate, repeatable, and compliant with international best practices.

SLOVENSKI STANDARD
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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
...