Foodstuffs - Determination of sucralose - High performance liquid chromatographic method

This European Standard specifies a method for the determination of sucralose in foodstuffs by high performance liquid chromatography (HPLC) by means of elution from a reversed-phased (RP) column using aqueous methanol, followed by RI detection [1]. This method has been validated in an inter-laboratory study via the analysis of sucralose (from 83 mg/kg to 737 mg/kg) in spiked samples of ketchup, mayonnaise, biscuits, yoghurt, instant beverage powder and sweets.
For further information on the validation results, see Annex A.

Lebensmittel - Bestimmung von Sucralose - Hochleistungsflüssigchromatographisches Verfahren

Diese Europäische Norm legt ein Verfahren zur Bestimmung von Sucralose in Lebensmitteln mit Hochleistungs¬flüssigchromatographie (HPLC) durch Elution von einer Umkehrphasensäule (RP Säule) unter Verwendung von wässrigem Methanol und anschließender RI Detektion [1] fest. Das Verfahren wurde in einem Ringversuch durch Analyse von Sucralose (von 83 mg/kg bis 737 mg/kg) in aufgestockten Proben von Ketchup, Mayonnaise, Keksen, Joghurt, sofort löslichem Getränkepulver und Süßwaren (Komprimaten) validiert.
Hinsichtlich weiterer Informationen zu den Validierungsergebnissen siehe Anhang A.

Produits alimentaires - Détermination du sucralose - Méthode par chromatographie liquide à haute performance

La présente Norme européenne spécifie une méthode de dosage du sucralose dans les produits alimentaires par chromatographie liquide à haute performance (CLHP), par élution sur une colonne en phase inverse (PI) en utilisant du méthanol aqueux, suivie d’une détection de l’indice de réfraction [1]. Cette méthode a été validée lors d’une étude interlaboratoires via l’analyse du sucralose (de 83 mg/kg à 737 mg/kg) dans des échantillons dopés de ketchup, mayonnaise, biscuits, yaourt, poudre pour boisson instantanée et bonbons.
Pour de plus amples informations sur les résultats de validation, voir l’Annexe A.

Živila - Določevanje sukraloze - Tekočinska kromatografija visoke ločljivosti

Ta evropski standard določa metodo za določevanje sukraloze v živilih s tekočinsko kromatografijo visoke ločljivosti (HPLC) prek elucije iz reverzne fazne (RP) kolone z uporabo vodnega metanola in pozneje z določanjem lomnega količnika. Ta metoda je bila validirana z medlaboratorijsko raziskavo prek analize sukraloze (od 83 mg/kg do 737 mg/kg) v primešanih vzorcih kečapa, majoneze, piškotov, jogurta, instantne pijače v prahu in sladkarij. Za več informacij o rezultatih validacije glejte dodatek C.

General Information

Status
Published
Public Enquiry End Date
14-Feb-2011
Publication Date
01-Aug-2012
ICS
67.050 - General methods of tests and analysis for food products
Technical Committee
KŽP - Agricultural food products
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
02-Jul-2012
Due Date
06-Sep-2012
Completion Date
02-Aug-2012
Ref Project
EN 16155:2012 - Foodstuffs - Determination of sucralose - High performance liquid chromatographic method

Overview

EN 16155:2012 (CEN) - Foodstuffs: Determination of sucralose - High performance liquid chromatographic method - specifies a validated laboratory procedure for measuring sucralose in food using reversed‑phase HPLC with aqueous methanol eluent and refractive index (RI) detection (ELSD may be used as an alternative). The method was validated in inter‑laboratory trials for spiked samples in a range of about 83 mg/kg to 737 mg/kg across matrices such as ketchup, mayonnaise, biscuits, yoghurt, instant beverage powder and sweets (see Annex A for validation data).

Key Topics and Requirements

  • Scope & principle: Extraction/dissolution of the food sample, optional protein clarification (modified Carrez), cleanup by C18 solid phase extraction (SPE), elution with methanol/water and HPLC quantification by external calibration.
  • Reagents & standards: Use analytical grade reagents and water complying with EN ISO 3696, sucralose primary standard (stock ≈ 1 000 mg/L) and daily standard solutions (e.g., 20–100 mg/L).
  • Apparatus: HPLC system with pump, injector, column oven, temperature‑controlled RI detector (or ELSD) and a C18 reversed‑phase column (e.g., 250 × 4 mm, 5 µm) with guard column; membrane filters (0.45 µm) and C18 SPE cartridges (500 mg).
  • Sample preparation: Homogenize; dissolve or extract (30 min, 40–60 °C or ultrasound); clarify protein matrices with modified Carrez reagents; condition SPE, load sample, wash and elute; filter eluate before HPLC.
  • Chromatography parameters: Typical settings used in validation - eluent methanol:water 1:3, column oven ~35 °C, RI detector ~30 °C, flow ≈ 1.2 mL/min, injection up to 100 µL (users adjust to instrument sensitivity).
  • Quantification & performance: External calibration (linear fit y = a + bx), integration of sucralose peak area and calculation using standard calibration function. Validation includes repeatability (r) and reproducibility (R) limits - e.g., ketchup r = 8.38 mg/kg, R = 18.16 mg/kg; biscuits r = 31.12 mg/kg, R = 130.89 mg/kg.

Applications and Users

  • Routine testing of sucralose content in food and beverage products for compliance, labeling verification and quality control.
  • Intended users: food testing laboratories, quality assurance teams in manufacturers, regulatory agencies, and method developers implementing official HPLC assays for sweetener analysis.
  • Useful for product development, market surveillance, and forensic food analysis where sucralose quantification is required.

Related Standards

  • Normative reference: EN ISO 3696:1995 (Water for analytical laboratory use).
  • Prepared by CEN/TC 275 “Food analysis - Horizontal methods”; national implementation by CEN members (see EN 16155:2012 foreword).

For full analytical details, validation statistics and reporting requirements, consult the complete EN 16155:2012 text and Annex A.

Frequently Asked Questions

SIST EN 16155:2012 is a standard published by the Slovenian Institute for Standardization (SIST). Its full title is "Foodstuffs - Determination of sucralose - High performance liquid chromatographic method". This standard covers: This European Standard specifies a method for the determination of sucralose in foodstuffs by high performance liquid chromatography (HPLC) by means of elution from a reversed-phased (RP) column using aqueous methanol, followed by RI detection [1]. This method has been validated in an inter-laboratory study via the analysis of sucralose (from 83 mg/kg to 737 mg/kg) in spiked samples of ketchup, mayonnaise, biscuits, yoghurt, instant beverage powder and sweets. For further information on the validation results, see Annex A.

This European Standard specifies a method for the determination of sucralose in foodstuffs by high performance liquid chromatography (HPLC) by means of elution from a reversed-phased (RP) column using aqueous methanol, followed by RI detection [1]. This method has been validated in an inter-laboratory study via the analysis of sucralose (from 83 mg/kg to 737 mg/kg) in spiked samples of ketchup, mayonnaise, biscuits, yoghurt, instant beverage powder and sweets. For further information on the validation results, see Annex A.

SIST EN 16155:2012 is classified under the following ICS (International Classification for Standards) categories: 67.050 - General methods of tests and analysis for food products. The ICS classification helps identify the subject area and facilitates finding related standards.

You can purchase SIST EN 16155:2012 directly from iTeh Standards. The document is available in PDF format and is delivered instantly after payment. Add the standard to your cart and complete the secure checkout process. iTeh Standards is an authorized distributor of SIST standards.

Standards Content (Sample)


2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von Sucralose - Hochleistungsflüssigchromatographisches VerfahrenProduits alimentaires - Détermination du sucralose - Méthode par chromatographie liquide à haute performanceFoodstuffs - Determination of sucralose - High performance liquid chromatographic method67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 16155:2012SIST EN 16155:2012en,fr,de01-september-2012SIST EN 16155:2012SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16155
April 2012 ICS 67.050 English Version
Foodstuffs - Determination of sucralose - High performance liquid chromatographic method
Produits alimentaires - Dosage du sucralose - Méthode par chromatographie liquide à haute performance
Lebensmittel - Bestimmung von Sucralose - Hochleistungsflüssigchromatographisches Verfahren This European Standard was approved by CEN on 25 February 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, 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 © 2012 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16155:2012: ESIST EN 16155:2012

Statistical data of the inter-laboratory trial .9Bibliography . 11 SIST EN 16155:2012

EN ISO 3696:1995, Water for analytical laboratory use — Specification and test methods (ISO 3696:1987) 3 Principle Depending on their consistency, the samples are either dissolved in water or diluted with water and, if appropriate, they are filtered or clarified with modified Carrez solutions. Afterwards, they are extracted on a solid phase extraction column and eluted with a mixture of methanol/water. The sucralose content is determined by high-performance liquid chromatography (HPLC) by means of elution from a reversed-phased (RP) column using aqueous methanol, followed by RI (refractive index) detection. Alternatively, an ELSD (Evaporative Light Scattering Detector) may be used. Quantification is performed by applying the external standard method. The content of sucralose in foodstuffs means identifying the content of 1,6-dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4-deoxy-α-D-galactopyranoside, as determined in accordance with the method described in this document.
4 Reagents Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696:1995, unless otherwise specified. Solvents shall be of the same quality used for HPLC analysis, unless otherwise specified. Commercially available solutions with equivalent properties to the reagents listed may be used.
4.1 Sucralose, (C12H19Cl3O8, MW: 397,63). 4.2 Potassium hexacyanoferrate(II), K4[Fe(CN)6] ⋅ 3H2O. 4.3 Zinc nitrate, Zn(NO3)2 ⋅ 6H2O. 4.4 Methanol for HPLC. 4.5 Stock solution (Suc ≈ 1 000 mg/l). Weigh approximately 100 mg of sucralose to the nearest 0,1 mg in a 100 ml volumetric flask; dissolve in a small amount of water and dilute to the calibration mark with water. Prepare the solution fresh every day. The water content and the purity of the standard substance shall be taken into consideration.
Prepare from the stock solution (4.5) at least five standard solutions by diluting in such a way that sucralose concentrations of e.g., 20 mg/l, 40 mg/l, 60 mg/l, 80 mg/l and 100 mg/l are obtained. Prepare these solutions fresh every day of the analysis. 4.7
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