EN 15911:2010

Foodstuffs - Simultaneous determination of nine sweeteners by high performance liquid chromatography and evaporative light scattering detection

This European Standard specifies a method for the simultaneous determination of nine sweeteners in beverages and canned or bottled fruits by high performance liquid chromatography (HPLC) with evaporative light scattering detection (HPLC-ELSD). This method has been validated in an interlaboratory study via the analysis of spiked samples on the following matrices:
-   acesulfame-K (ACS-K) (from 38,3 mg/l to 383,5 mg/l) in beverages and (from 38,4 mg/kg to 391,3 mg/kg) in canned fruits;
-   alitame (ALI) (from 31,1 mg/l to 114,5 mg/l) in beverages and (from 36 mg/kg to 175,2 mg/kg) in canned fruits;
-   aspartame (ASP) (from 38,1 mg/l to 702 mg/l) in beverages and (from 37,2 mg/kg to 1 120,2 mg/kg) in canned fruits;
-   cyclamic acid (CYC) (from 28,3 mg/l to 307,2 mg/l) in beverages and (from 27,5 mg/kg to 1 100,6 mg/kg) in canned fruits;
-   dulcin (DUL) (from 55,0 mg/l to 115,1 mg/l) in beverages and (from 49,8 mg/kg to 172,6 mg/kg) in canned fruits;
-   neotame (NEO) (from 37,6 mg/l to 115,3 mg/l) in beverages and (from 37,3 mg/kg to 173,7 mg/kg) in canned fruits;
-   neohesperidine dihydrochalcone (NHDC) (from 31,4 mg/l to 59,3 mg/l) in beverages and (from 35,3 mg/kg to 59,3 mg/kg) in canned fruits;
-   saccharin (SAC) (from 36,2 mg/l to 87,6 mg/l) in beverages and (from 44,3 mg/kg to 235,3 mg/kg) in canned fruits;
-   sucralose (SCL) (from 36,8 mg/l to 346,8 mg/l) in beverages and (from 35,3 mg/kg to 462,4 mg/kg) in canned fruits.
For further information on the validation results see Annex C.
NOTE   The method has been fully validated [1] through collaborative trial, according to the IUPAC Harmonised Protocol [2], on analyte-matrix combinations of all nine sweeteners in beverages and canned or bottled fruits.

Lebensmittel - Gleichzeitige Bestimmung von neun Süßungsmitteln mit Hochleistungs-Flüssigchromatographie und Verdampfungs-Lichtstreu-Detektion

Diese Europäische Norm legt ein Verfahren zur gleichzeitigen Bestimmung von neun Intensivsüßungsmitteln in
Getränken und in Dosen konserviertem oder eingemachtem Obst durch Hochleistungs-Flüssigchromatographie
mit Verdampfungs-Lichtstreu-Detektion (HPLC-ELSD) fest. Das Verfahren wurde in einem Ringversuch durch
die Analyse von aufgestockten Proben an den folgenden Matrices validiert:
⎯ Acesulfam-K (ACS-K) (von 38,3 mg/l bis 383,5 mg/l) in Getränken und (von 38,4 mg/kg bis 391,3 mg/kg)
in Dosenobst;
⎯ Alitam (ALI) (von 31,1 mg/l bis 114,5 mg/l) in Getränken und (von 36 mg/kg bis 175,2 mg/kg) in
Dosenobst;
⎯ Aspartam (ASP) (von 38,1 mg/l bis 702 mg/l) in Getränken und (von 37,2 mg/kg bis 1 120,2 mg/kg) in
Dosenobst;
⎯ Cyclohexansulfamidsäure (CYC) (von 28,3 mg/l bis 307,2 mg/l) in Getränken und (von 27,5 mg/kg bis
1 100,6 mg/kg) in Dosenobst;
⎯ Dulcin (DUL) (von 55,0 mg/l bis 115,1 mg/l) in Getränken und (von 49,8 mg/kg bis 172,6 mg/kg) in
Dosenobst;
⎯ Neotam (NEO) (von 37,6 mg/l bis 115,3 mg/l) in Getränken und (von 37,3 mg/kg bis 173,7 mg/kg) in
Dosenobst;
⎯ Neohesperidin-Dihydrochalcon (NHDC) (von 31,4 mg/l bis 59,3 mg/l) in Getränken und (von 35,3 mg/kg
bis 59,3 mg/kg) in Dosenobst;
⎯ Saccharin (SAC) (von 36,2 mg/l bis 87,6 mg/l) in Getränken und (von 44,3 mg/kg bis 235,3 mg/kg) in
Dosenobst;
⎯ Sucralose (SCL) (von 36,8 mg/l bis 346,8 mg/l) in Getränken und (von 35,3 mg/kg bis 462,4 mg/kg) in
Dosenobst.
Weitere Informationen zu den Validierungsergebnissen siehe Abschnitt 8 und Anhang C.
ANMERKUNG Das Verfahren wurde durch einen Ringversuch [1] nach dem IUPAC-Harmonisierungsprotokoll [2] an
Analyt-Matrix-Kombinationen aller neun Süßungsmittel in Getränken und in Dosen konserviertem oder eingemachtem
Obst vollständig validiert.

Denrées alimentaires - Détermination simultanée de neuf édulcorants par chromatographie liquide haute performance et détection à diffusion de lumière

La présente Norme européenne spécifie une méthode de dosage simultané de neuf édulcorants dans des
boissons et des fruits en conserve ou en bouteille par chromatographie liquide haute performance (CLHP)
avec détection à diffusion de lumière (CLHP-ELSD). La présente méthode a été validée dans le cadre d'une
étude interlaboratoires par le biais de l'analyse d'échantillons supplémentés sur les matrices suivantes :
- acésulfame-K (ACS-K) (de 38,3 mg/l à 383,5 mg/l) dans les boissons et (de 38,4 mg/kg à 391,3 mg/kg)
dans les fruits en conserve ;
- alitame (ALI) (de 31,1 mg/l à 114,5 mg/l) dans les boissons et (de 36 mg/kg à 175,2 mg/kg) dans les fruits
en conserve ;
- aspartame (ASP) (de 38,1 mg/l à 702 mg/l) dans les boissons et (de 37,2 mg/kg à 1 120,2 mg/kg) dans
les fruits en conserve ;
- acide cyclamique (CYC) (de 28,3 mg/l à 307,2 mg/l) dans les boissons et (de 27,5 mg/kg à
1 100,6 mg/kg) dans les fruits en conserve ;
- dulcine (DUL) (de 55,0 mg/l à 115,1 mg/l) dans les boissons et (de 49,8 mg/kg à 172,6 mg/kg) dans les
fruits en conserve ;
- néotame (NEO) (de 37,6 mg/l à 115,3 mg/l) dans les boissons et (de 37,3 mg/kg à 173,7 mg/kg) dans les
fruits en conserve ;
- néohespéridine dihydrochalcone (NHDC) (de 31,4 mg/l à 59,3 mg/l) dans les boissons et (de 35,3 mg/kg
à 59,3 mg/kg) dans les fruits en conserve ;
- saccharine (SAC) (de 36,2 mg/l à 87,6 mg/l) dans les boissons et (de 44,3 mg/kg à 235,3 mg/kg) dans les
fruits en conserve ;
- sucralose (SAC) (de 36,8 mg/l à 346,8 mg/l) dans les boissons et (de 35,3 mg/kg à 462,4 mg/kg) dans les
fruits en conserve.
Pour plus d'informations sur la validation, voir l'Article 8 et l'Annexe C.
NOTE La méthode a été intégralement validée [1] dans le cadre d’une étude interlaboratoires, conformément au
Protocole harmonisé IUPAC [2], sur des combinaisons analyte/matrice des neuf édulcorants dans des boissons et des
fruits en conserve ou en bouteille.

Živila - Simultano določevanje devetih sladil z metodo tekočinske kromatografije visoke ločljivosti in z detekcijo disperzije svetlobe (ELS)

Ta evropski standard določa metodo za simultano določevanje devetih sladil v pijačah in konzerviranega ali stekleničenega sadja s tekočinsko kromatografijo visoke ločljivosti (HPLC) z detekcijo disperzije svetlobe (HPLC-ELSD). Ta metoda je bila potrjena v medlaboratorijski študiji z analizo vzorcev z internimi dodatki na naslednjih matricah:
– acusulfam-K (ACS-K) (od 38,3 mg/l do 383,5 mg/l) v pijačah in (od 38,4 mg/kg do 391,3 mg/kg) v konzerviranem sadju;
– alitam (ALI) (od 31,1 mg/l do 114,5 mg/l) v pijačah in (od 36 mg/kg do 175,2 mg/kg) v konzerviranem sadju;
– aspartam (ASP) (od 38,1 mg/l do 702 mg/l) v pijačah in (od 37,2 mg/kg do 1.120,2 mg/kg) v konzerviranem sadju;
– ciklaminska kislina (CYC) (od 28,3 mg/l do 307,2 mg/l) v pijačah in (od 27,5 mg/kg do 1.100,6 mg/kg) v konzerviranem sadju;
– dulcin (DUL) (od 55,0 mg/l do 115,1 mg/l) v pijačah in (od 49,8 mg/kg do 172,6 mg/kg) v konzerviranem sadju;
– neotam (NEO) (od 37,6 mg/l do 115,3 mg/l) v pijačah in (od 37,3 mg/kg do 173,7 mg/kg) v konzerviranem sadju;
– neohesperidin dihidrokalcon (NHDC) (od 31,4 mg/l do 59,3 mg/l) v pijačah in (od
35,5 mg/kg do 59,3 mg/kg) v konzerviranem sadju;
– saharin (SAC) (od 36,2 mg/l do 87,6 mg/l) v pijačah in (od 44,3 mg/kg do 235,3 mg/kg) v konzerviranem sadju;
– sukraloza (SCL) (od 36,8 mg/l do 346,8 mg/l) v pijačah in (od 35,3 mg/kg do 462,4 mg/kg) v konzerviranem sadju. Za dodatne informacije o potrjevanju glej klavzulo 8 in dodatek C.

General Information

• Status: Published
• Publication Date: 19-Oct-2010
• ICS: 67.180.10 - Sugar and sugar products
• Technical Committee: CEN/TC 275 - Food analysis - Horizontal methods
• Drafting Committee: CEN/TC 275/WG 2 - Sweeteners
• Current Stage: 9093 - Decision to confirm - Review Enquiry
• Completion Date: 02-Jul-2021
• Ref Project: SIST EN 15911:2011 - Foodstuffs - Simultaneous determination of nine sweeteners by high performance liquid chromatography and evaporative light scattering detection

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Gleichzeitige Bestimmung von neun Süßungsmitteln mit Hochleistungs-Flüssigchromatographie und Verdampfungs-Lichtstreu-DetektionDenrées alimentaires - Dosage simultané de neuf édulcorants par chromatographie liquide haute performance et détection à diffusion de lumière par évaporationFoodstuffs - Simultaneous determination of nine sweeteners by high performance liquid chromatography and evaporative light scattering detection67.180.10Sladkor in sladkorni izdelkiSugar and sugar productsICS:Ta slovenski standard je istoveten z:EN 15911:2010SIST EN 15911:2011en,fr,de01-marec-2011SIST EN 15911:2011SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 15911
October 2010 ICS 67.180.10 English Version
Foodstuffs - Simultaneous determination of nine sweeteners by high performance liquid chromatography and evaporative light scattering detection
Denrées alimentaires - Détermination simultanée de neuf édulcorants par chromatographie liquide haute performance et détection à diffusion de lumière
Lebensmittel - Gleichzeitige Bestimmung von neun Süßungsmitteln mit Hochleistungs-Flüssigchromatographie und Verdampfungs-Lichtstreu-Detektion This European Standard was approved by CEN on 18 September 2010.
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 and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2010 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 15911:2010: ESIST EN 15911:2011

Table A.1
Suitable method conditions . 16Annex B (informative)
Examples of chromatograms . 17Annex C (informative)
Precision data . 19Annex D (informative)
Present EU limits for the nine sweeteners . 29Bibliography . 30 SIST EN 15911:2011

98,0 %. 4.8 Sodium cyclamate, w ≥
99,0 %. 4.9 Sucralose, w ≥ 99,0 %. 4.10 Formic acid, HCOOH for HPLC. 4.11 Triethylamine, (C2H5)3N, w ≥ 99,5 %. 4.12 Methanol, for HPLC. 4.13 Acetone, for HPLC. 4.14 Buffer solution (pH = 4,5). Dissolve 4 ml of formic acid (4.10) in 5 l of water. Adjust to pH 4,5 with approximately 12,5 ml triethylamine (4.11). 4.15 HPLC mobile phase A, methanol/buffer solution/acetone 69:24:7 (v/v/v). Mix 690 ml of methanol (4.12) with 240 ml of buffer solution (4.14) and with 70 ml of acetone (4.13). Degas by sonication for 10 min. 4.16 HPLC Mobile phase B, methanol/buffer solution/acetone 11:82:7 (v/v/v). Mix 110 ml of methanol (4.12) with 820 ml of buffer solution (4.14) and with 70 ml of acetone (4.13). Degas by sonication for 10 min. 4.17 Mixed stock solution, containing ACS-K, ALI, ASP, CYC-Na, DUL, NEO, NHDC, SAC-Na and SCL; mass concentration ρ(sweetener i) = 30 µg/ml to 250 µg/ml. Prepare a mixed stock solution of all nine sweeteners by weighing the given masses of the individual sweetener standards (Table 1) first into a 100 ml beaker and dissolving them in 50 ml of methanol/water (1:1) until complete dissolution. Then transfer the obtained solution quantitatively into a 500 ml volumetric flask and make up to the mark with the buffer solution (4.14). Mix thoroughly by sonication until complete dissolution. SIST EN 15911:2011

NOTE In case of cyclamic acid and saccharin, their sodium salts are used, since they are either not available in free form or poorly soluble. The final concentrations of the individual sweeteners in micrograms per millilitre in the mixed stock solution have to be calculated by using the actually weighed masses. 4.18 Standard solutions. From the mixed stock solution (4.17) prepare a series of standard solutions containing the sweeteners at levels fitting appropriate limits, e.g. the highest concentration of the calibration shall be at least equivalent to 125 % of the given limits, such as those in Commission Directives [3], [4], [5] (see Table D.1), whilst taking the dilution steps within the procedure into account (see Table 2). For sweeteners not authorised by the current EU legislation (ALI, DUL and NEO) fictitious maximum usable dosages (MUD) are assumed at approximately 200 mg/l or 200 mg/kg. The user of the standard has to check whether the limits in Table D.1 are still valid. If not, the mass concentration of the standard substance in the calibration solution shall be adjusted to meet the current MUDs. NOTE The present procedure is simplified by preparing one calibration series for both food matrices. The described calibration series is fitted to canned fruits as the MUDs for canned fruits are in some cases higher than the MUDs for beverages. In case only the latter matrix is analysed the calibration series can be fitted to the MUDs of beverages. Pipette the following volumes (see Table 2) from the mixed stock solution (4.17) into appropriate volumetric flasks (10 ml to 50 ml) and make up to the mark with buffer solution (4.14) and shake thoroughly. SIST EN 15911:2011

Table 3 details the concentration of sweetener i in each calibration standard following preparation described in Table 2. If not all of the sweeteners covered by this standard are the subject of analysis in routine use of the method, when applied to a particular set of samples consideration may be given to reduce the levels of the calibration solutions used for those samples. Table 3 — Concentration of sweetener i in the individual standard solutions Sweetener Calibration solution mg/ml
1 2 3 4 5 6 7 8 ACS-K 90,0 72,0 54,0 36,0 18,0 10,8 5,4 2,7 a ALI 50,0 40,0 30,0 20,0 10,0 6,0 3,0 a 1,5 a ASP 250,0 200,0 150,0 100,0 50,0 30,0 15,0 7,5 CYC 249,4 199,5 149,7 99,8 49,9 29,9 15,0 7,5 DUL 50,0 40,0 30,0 20,0 10,0 6,0 a 3,0 a 1,5 a NEO 50,0 40,0 30,0 20,0 10,0 6,0 3,0 a 1,5 a NHDC 30,0 24,0 18,0 12,0 6,0 3,6 a 1,8 a 0,9 a SAC 53,2 42,5 31,9 21,3 10,6 6,4 3,2 a 1,6 a SCL 100,0 80,0 60,0 40,0 20,0 12,0 6,0 3,0 a a The concentration level might be below the limit of quantification (LOQ). If yes, the result obtained by HPLC analysis is not included in the construction of the calibration graph, e.g. in case of ACS-K a seven point calibration is performed, ignoring the result obtained for calibration solution 8. 5 Apparatus and equipment Usual laboratory apparatus and, in particular, the following: 5.1 Common laboratory glassware, such as graduated cylinders, volumetric pipettes, glass beakers. 5.2 Analytical balance, capable of weighing to 0,01 mg. 5.3 Laboratory balance, capable of weighing to 0,01 g. 5.4 Positive displacement pipette, or equ
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