SIST EN 16466-2:2013

Vinegar - Isotopic analysis of acetic acid and water - Part 2: 13C-IRMS analysis of acetic acid

This European standard specifies an isotopic method to control the authenticity of vinegar. This method is applicable on acetic acid of vinegar (from cider, alcohol, wine, etc.) in order to characterise the botanical origin of acetic acid and to detect adulterations of vinegar using synthetic acetic acid or acetic acid from not allowed origin (together with the method described in FprEN 16466-1).
The isotopic analysis of the extracted acetic acid by 13C-IRMS is based on a similar method already normalised for wine analysis [2].

Essig - Isotopenanalyse von Essigsäure und Wasser - Teil 2: 13C-IRMS-Analyse von Essigsäure

Diese Europäische Norm legt ein isotopenanalytisches Verfahren zur Überprüfung der Authentizität von Essig fest. Dieses Verfahren ist anwendbar auf Essigsäure aus Essig (aus Apfelwein, Alkohol, Wein usw.), um den botanischen Ursprung der Essigsäure zu charakterisieren und um Verfälschungen von Essig durch Verwendung synthetischer Essigsäure oder Essigsäure aus einem nicht zulässigen Ursprung nachzuweisen (in Kombination mit dem in EN 16466 1 beschriebenen Verfahren).
Die Isotopenanalyse der extrahierten Essigsäure durch 13C IRMS beruht auf einem ähnlichen Verfahren, das bereits für die Weinanalyse genormt wurde [2].

Vinaigre - Analyse isotopique de l'acide acétique et de l'eau - Partie 2 : Analyse SMRI-13C de l'acide acétique

La présente Norme européenne spécifie une méthode isotopique de contrôle de l’authenticité du vinaigre. La présente méthode s’applique à l’acide acétique de vinaigre (de cidre, d’alcool, de vin, etc.) afin de caractériser l’origine botanique de l’acide acétique et de détecter des adultérations du vinaigre utilisant de l’acide acétique synthétique ou de l’acide acétique d’une origine non autorisée (en complément de la méthode décrite dans l’EN 16466-1).
L’analyse isotopique par SMRI-13C de l’acide acétique extrait est fondée sur une méthode similaire à celle de l’analyse du vin, déjà normalisée [2].

Kis - Izotopska analiza ocetne kisline in vode - 2. del: 13C-IRMS-analiza ocetne kisline

Ta evropski standard določa izotopsko metodo za nadzorovanje pristnosti kisa. Ta metoda se uporablja za ocetno kislino v kisu (iz mošta, alkohola, vina itd.), da se opredelijo značilnosti botaničnega izvora ocetne kisline in zazna ponarejanje kisa z uporabo sintetične ocetne kisline ali ocetne kisline nedovoljenega izvora (skupaj z metodo iz standarda EN 16466-1). Izotopska analiza ekstrahirane ocetne kisline s 13C-IRMS temelji na podobni metodi, ki je že normalizirana za analizo vina.

General Information

• Status: Published
• Publication Date: 21-May-2013
• ICS: 67.220.20 - Food additives
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 25-Apr-2013
• Due Date: 30-Jun-2013
• Completion Date: 22-May-2013
• Ref Project: EN 16466-2:2013 - Vinegar - Isotopic analysis of acetic acid and water - Part 2: 13C-IRMS analysis of acetic acid

Overview

SIST EN 16466-2:2013 is a European standard developed by CEN that specifies a precise isotopic analysis method for verifying the authenticity of vinegar. This standard focuses on the 13C Isotope Ratio Mass Spectrometry (13C-IRMS) analysis of acetic acid extracted from different vinegars including cider, alcohol, wine vinegars, and others. The method allows for characterization of the botanical origin of acetic acid and detects adulterations, particularly the use of synthetic acetic acid or acetic acid from prohibited sources.

The standard supports quality control and authenticity verification in vinegar production by leveraging isotopic techniques already established for wine analysis. It is part of a comprehensive European framework for vinegar authenticity composed of several related parts analyzing different isotopes.

Key Topics

• Scope and Applicability
  The method is applicable to vinegar acetic acid derived from natural fermentation sources and excludes non-permitted industrial or synthetic origins.

• Principle of Analysis
  Acetic acid is extracted from vinegar samples via liquid-liquid extraction using diethyl ether or similar solvents. The extracted acetic acid is purified by distillation, then combusted to CO2. The 13C/12C isotope ratio is measured by IRMS to determine the isotopic signature.

• Reagents and Apparatus

  • Use of diethyl ether, CO2, helium, oxygen gases of high purity
  • Equipment includes liquid-liquid extractors, distillation apparatus (spinning band or Vigreux columns), elemental analyzers, and isotope ratio mass spectrometers equipped with triple collectors and automatic samplers.

• Test Procedure
  Detailed steps cover extraction, purification, combustive conversion, and isotope ratio measurement, with an emphasis on sample preparation and instrument calibration for precision.

• Data Analysis and Reporting
  Results are expressed in delta notation (δ13C‰) relative to an internationally accepted reference (V-PDB). Linear interpolation corrects instrumental drift using working standards measured throughout the sample batch. The standard mandates precise reporting criteria including sample identification, test conditions, and any deviations from the method.

• Performance and Precision
  Collaborative studies show excellent repeatability (0.45‰) and reproducibility (0.91‰) in δ13C measurements, confirming method reliability for food authenticity control.

Applications

• Vinegar Authenticity Control
  Ensures vinegars marketed comply with natural fermentation requirements and are free from synthetic acetic acid adulteration. Essential for regulatory compliance and consumer protection.

• Botanical Origin Verification
  Differentiates vinegars derived from various botanical sources such as grapes, apples, or cereals based on characteristic 13C isotopic ratios, aiding provenance labeling and fraud detection.

• Food Quality and Safety Laboratories
  Provides a standardized analytical protocol for commercial and regulatory laboratories performing vinegar authenticity testing and certification.

• Wine Industry Utilization
  Uses isotopic methodologies aligned with standardized wine ethanol testing, facilitating consistent cross-product authenticity assessment.

Related Standards

• EN 16466-1
  Details 2H-NMR isotopic analysis of acetic acid, complementing the 13C-IRMS method in authentication procedures.

• EN 16466-3
  Specifies 18O-IRMS analysis of water in vinegar, contributing additional isotopic signatures for comprehensive authenticity evaluation.

• EN 13188 and EN 13189
  Define vinegar and food-grade acetic acid product requirements, including origin and production process criteria aiding in enforcement of standards.

• EU Regulations 479/2008 and 491/2009
  Provide legal definitions and specifications for wine vinegars within the European market, reinforcing the technical requirements for vinegar authenticity tested under EN 16466-2.

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The SIST EN 16466-2:2013 standard is a critical tool for ensuring the integrity and transparency of vinegar products across the European Union, supporting trustworthy labeling and safeguarding against adulteration through robust, scientifically validated isotopic analysis.

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Kis - Izotopska analiza ocetne kisline in vode - 2. del: 13C-IRMS-analiza ocetne kislineEssig - Isotopenanalyse von Essigsäure und Wasser - Teil 2: 13C-IRMS-Analyse von EssigsäureVinaigre - Analyse isotopique de l'acide acétique et de l'eau - Partie 2 : Analyse SMRI-13C de l'acide acétiqueVinegar - Isotopic analysis of acetic acid and water - Part 2: 13C-IRMS analysis of acetic acid67.220.20Dodatki jedemFood additivesICS:Ta slovenski standard je istoveten z:EN 16466-2:2013SIST EN 16466-2:2013en,fr,de01-junij-2013SIST EN 16466-2:2013SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16466-2
January 2013 ICS 67.220.10
English Version
Vinegar - Isotopic analysis of acetic acid and water - Part 2:
13C-IRMS analysis of acetic acid
Vinaigre - Analyse isotopique de l'acide acétique et de l'eau - Partie 2 : Analyse SMRI-13C de l'acide acétique
Essig - Isotopenanalyse von Essigsäure und Wasser - Teil 2: 13C-IRMS-Analyse von Essigsäure This European Standard was approved by CEN on 3 November 2012.
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
Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16466-2:2013: E SIST EN 16466-2:2013

Results of the collaborative study (2009) . 9 Bibliography. 10
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 July 2013, and conflicting national standards shall be withdrawn at the latest by July 2013. 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. The European standard, Vinegar — Isotopic analysis of acetic acid and water, consists of the following parts:  Part 1: 2H-NMR analysis of acetic acid;  Part 2: 13C-IRMS analysis of acetic acid;  Part 3: 18O-IRMS analysis of water. According to the CEN/CENELEC Internal Regulations, the national standards organisations 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.
a) transformation of sugars to ethanol and
b) transformation of ethanol to acetic acid.
Conversely EN 13189 defines acetic acid as "Product made from materials of non-agricultural origin".
Wine vinegar is defined by the European Regulations 479/2008 and 491/2009 as the product obtained exclusively from the acetous fermentation of wine, which is in turn defined as the product exclusively obtained from the alcoholic fermentation of fresh grapes, whether crushed or not, or of grape must.
In all types of vinegar, both the ethanol and the acetic acid should be obtained by a biotechnological process, and the use of acetic acids obtained from either petroleum derivatives or the pyrolysis of wood is not permitted according to the above definitions.
The isotopic analysis of acetic acid extracted from vinegar by 2H-SNIF-NMR and 13C-IRMS enables the distinction of grape origin from other sources, such as beet, cane, malt, apple and synthesis [1].
4 Reagents All reagents and consumables used shall meet stated requirements of the used method/apparatus (as specified by the manufacturer). However, all reagents and consumables can be replaced by items with similar performance. 4.1 Diethyl ether For analysis. 4.2 Carbon dioxide For analysis, used as secondary reference gas for the determination of 13C/12C ratio. Purity 5.2 minimum. 4.3 Helium For analysis. Purity 5.6 minimum. 4.4 Oxygen For analysis. Purity 5.0 minimum. SIST EN 16466-2:2013

5.1.1 Liquid-liqui
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