EN 12823-1:2014
(Main)Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 1: Measurement of all-E-retinol and 13-Z-retinol
Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 1: Measurement of all-E-retinol and 13-Z-retinol
This European Standard specifies a method for the determination of vitamin A in foodstuffs by high performance liquid chromatography (HPLC). This method has been validated in an interlaboratory study with samples of margarine and milk powder with all-E-retinol levels ranging from 653 µg/100 g to 729 µg/100 g and with 13-Z-retinol levels ranging from 30 µg/100 g to 39 µg/100 g. The determination of vitamin A content is carried out by the measurement of all-E-retinol, 13-Z-retinol and β-carotene. This part covers the measurement of all-E-retinol and 13-Z-retinol.
The extract obtained after saponification in this method can be used for the determination of β-carotene, as described in EN 12823 2:2000 Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 2: Measurements of β-carotene. In this case, the saponification temperature should preferably not exceed 80 °C in order to prevent isomerisation and oxidation of β-carotene.
Lebensmittel - Bestimmung von Vitamin A mit Hochleistungs-Flüssigchromatographie - Teil 1: Bestimmung von all-E-Retinol und 13-Z-Retinol
Diese Europäische Norm legt ein Verfahren zur Bestimmung von Vitamin A in Lebensmitteln mit Hochleistungs-Flüssigchromatographie (HPLC) fest. Diese Bestimmung wurde in einem Ringversuch an Margarine und Milchpulver mit 653 μg/100 g bis 729 μg/100 g all-(E)-Retinol und mit 30 μg/100 g bis 39 μg/100 g 13-Z-Retinol validiert. Die Bestimmung des Vitamin A-Gehaltes erfolgt durch Bestimmung von all-E-Retinol,13-Z-Retinol und von -Carotin. Dieser Teil enthält die Bestimmung von all-E-Retinol und 13 Z-Retinol.
Der in diesem Verfahren nach der Verseifung gewonnene Extrakt kann für die Bestimmung des -Carotins verwendet werden, wie in EN 12823-2:2000 Lebensmittel — Bestimmung von Vitamin A mit Hochleistungs-Flüssigchromatographie — Teil 2: Bestimmung von β-Carotin beschrieben. In diesem Fall sollte die Verseifungstemperatur 80 °C möglichst nicht überschreiten, um eine Isomerisierung und Oxidation von Carotin zu vermeiden.
Produits alimentaires - Détermination de la teneur en vitamine A par chromatographie liquide haute performance - Partie 1: Dosage du tout-E-rétinol et du 13-Z-rétinol
La présente Norme européenne spécifie une méthode de dosage de la vitamine A présente dans les produits alimentaires par chromatographie liquide haute performance (CLHP). Cette méthode a été validée lors d’un essai interlaboratoires portant sur des échantillons de margarine et de lait en poudre avec des concentrations en tout-E-rétinol comprises entre 653 µg/100 g et 729 µg/100 g et des concentrations en 13 Z-rétinol comprises entre 30 µg/100 g et 39 µg/100 g. La détermination de la teneur en vitamine A s’effectue par dosage du tout-E-rétinol, du 13-Z-rétinol et du β-carotène. La présente partie concerne le dosage du
tout-E-rétinol et du 13-Z-rétinol.
L'extrait obtenu après saponification par le biais de cette méthode peut être utilisé pour le dosage du
β-carotène, comme décrit dans l’EN 12823-2:2000, Produits alimentaires – Dosage de la vitamine A par chromatographie liquide haute performance – Partie 2 : Dosage du β-carotène. Dans ce cas, il convient de préférence que la température de saponification ne dépasse pas 80 °C pour éviter l’isomérisation et l’oxydation du β-carotène.
Živila - Določevanje vitamina A s tekočinsko kromatografijo visoke ločljivosti - 1. del: Merjenje vseh trans-retinolov in 13-cis retinola
Standard EN 12823-1 navaja metodo za določanje vitamina A v živilih s tekočinsko kromatografijo visoke ločljivosti (HPLC). Ta metoda je bila validirana z medlaboratorijsko študijo prek analize vzorcev margarine in mleka v prahu z nivoji vseh E-retinolov od 653 μg/100 g do 729 μg/100 g in nivoji 13-Z-retinola od 30 μg/100 g do 39 μg/100 g. Določanje deleža vitamina A se izvaja z merjenjem vseh E-retinolov, 13-Z-retinola in β-karotena. Ta del zajema merjenje vseh E-retinolov in 13-Z-retinola. S to metodo po umiljenju pridobljeni ekstrakt se lahko uporabi za določanje β-karotena, kot je opisano v standardu EN 12823-2:2000, Živila - Določevanje vitamina A s tekočinsko kromatografijo visoke ločljivosti - 2. del: Merjenje β-karotena. V tem primeru naj temperatura umiljenja po možnosti ne presega 80° C, da se prepreči izomerizacija in oksidacija β-karotena.
General Information
Relations
Overview
EN 12823-1:2014 (CEN) describes a validated analytical method for the determination of vitamin A in foodstuffs by high performance liquid chromatography (HPLC). Part 1 covers the measurement of all‑E‑retinol (all‑trans retinol) and 13‑Z‑retinol (13‑cis retinol). The method was validated in an interlaboratory study using margarine and milk powder with all‑E levels ~653–729 µg/100 g and 13‑Z levels ~30–39 µg/100 g. The procedure includes sample saponification, extraction, chromatographic separation and quantification by UV or fluorescence detection.
Key Topics and Requirements
- Analytical scope: Measurement of all‑E‑retinol and 13‑Z‑retinol by HPLC; β‑carotene measurement is covered in EN 12823‑2.
- Sample preparation: Saponification (reflux or alternative conditions), antioxidant use, solvent extraction and concentration of extracts for HPLC analysis.
- Saponification guidance: Typical conditions 15–45 min at 80–100 °C; if the extract will be used for β‑carotene determination, saponification preferably should not exceed 80 °C to avoid isomerisation/oxidation.
- Chromatography: Normal‑phase silica columns (examples: LiChrospher® Si 60 / LiChrosorb® Si 60) and mobile phases such as n‑hexane/2‑propanol or iso‑octane/2‑propanol; baseline resolution of all‑E and 13‑Z is required.
- Detection & quantification: UV‑VIS or fluorescence detection; identification by retention time and quantification by external standard using peak areas or heights.
- Standards & stability: Preparation of all‑E and 13‑Z stock/standard solutions; spectrometric concentration and purity checks (absorbance maxima around 325–326 nm for all‑E; molar extinction coefficients provided). Store vitamin A standards protected from light, under inert gas, typically at −20 °C.
- Quality reagents & water: Use analytical grade reagents and water meeting EN ISO 3696.
Applications and Who Uses This Standard
- Food testing laboratories performing routine or regulatory analysis of vitamin A in fortified and unfortified food matrices (dairy, margarine, powders).
- Quality control teams in food manufacturing verifying label claims and nutritional content.
- Regulatory agencies and research organizations conducting intercomparison studies and method validation.
- Analysts seeking a harmonized, CEN‑endorsed HPLC procedure for vitamin A determination in foodstuffs.
Practical Value
- Provides a harmonized, validated HPLC workflow ensuring comparable results across laboratories.
- Offers practical guidance for sample handling (light/oxygen protection, nitrogen blanketing), chromatographic conditions, and standard preparation to minimize isomerisation and degradation.
- Facilitates compliance with nutrition labeling and food safety programs.
Related Standards
- EN 12823‑2:2000 - Determination of vitamin A by HPLC - Part 2: Measurement of β‑carotene (useful when the same extract is analyzed for provitamin A carotenoids).
- EN ISO 3696 - Water for analytical laboratory use.
Frequently Asked Questions
EN 12823-1:2014 is a standard published by the European Committee for Standardization (CEN). Its full title is "Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 1: Measurement of all-E-retinol and 13-Z-retinol". This standard covers: This European Standard specifies a method for the determination of vitamin A in foodstuffs by high performance liquid chromatography (HPLC). This method has been validated in an interlaboratory study with samples of margarine and milk powder with all-E-retinol levels ranging from 653 µg/100 g to 729 µg/100 g and with 13-Z-retinol levels ranging from 30 µg/100 g to 39 µg/100 g. The determination of vitamin A content is carried out by the measurement of all-E-retinol, 13-Z-retinol and β-carotene. This part covers the measurement of all-E-retinol and 13-Z-retinol. The extract obtained after saponification in this method can be used for the determination of β-carotene, as described in EN 12823 2:2000 Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 2: Measurements of β-carotene. In this case, the saponification temperature should preferably not exceed 80 °C in order to prevent isomerisation and oxidation of β-carotene.
This European Standard specifies a method for the determination of vitamin A in foodstuffs by high performance liquid chromatography (HPLC). This method has been validated in an interlaboratory study with samples of margarine and milk powder with all-E-retinol levels ranging from 653 µg/100 g to 729 µg/100 g and with 13-Z-retinol levels ranging from 30 µg/100 g to 39 µg/100 g. The determination of vitamin A content is carried out by the measurement of all-E-retinol, 13-Z-retinol and β-carotene. This part covers the measurement of all-E-retinol and 13-Z-retinol. The extract obtained after saponification in this method can be used for the determination of β-carotene, as described in EN 12823 2:2000 Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 2: Measurements of β-carotene. In this case, the saponification temperature should preferably not exceed 80 °C in order to prevent isomerisation and oxidation of β-carotene.
EN 12823-1:2014 is classified under the following ICS (International Classification for Standards) categories: 67.040 - Food products in general; 67.050 - General methods of tests and analysis for food products. The ICS classification helps identify the subject area and facilitates finding related standards.
EN 12823-1:2014 has the following relationships with other standards: It is inter standard links to EN 12823-1:2000. Understanding these relationships helps ensure you are using the most current and applicable version of the standard.
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Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von Vitamin A mit Hochleistungs-Flüssigchromatographie - Teil 1: Bestimmung von all-trans-Retinol und 13-cis-RetinolProduits alimentaires - Dosage de la vitamine A par chromatographie liquide haute performance - Partie 1: Dosage du tout-trans-rétinol et du 13-cis-rétinolFoodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 1: Measurement of all-trans-retinol and 13-cis-retinol67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 12823-1:2014SIST EN 12823-1:2014en,fr,de01-julij-2014SIST EN 12823-1:2014SLOVENSKI
STANDARDSIST EN 12823-1:20001DGRPHãþD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 12823-1
May 2014 ICS 67.050 Supersedes EN 12823-1:2000English Version
Foodstuffs - Determination of vitamin A by high performance liquid chromatography - Part 1: Measurement of all-E-retinol and 13-Z-retinol
Produits alimentaires - Détermination de la teneur en vitamine A par chromatographie liquide haute performance - Partie 1: Dosage du tout-E-rétinol et du 13-Z-rétinol
Lebensmittel - Bestimmung von Vitamin A mit Hochleistungs-Flüssigchromatographie - Teil 1: Bestimmung von all-E-Retinol und 13-Z-Retinol This European Standard was approved by CEN on 24 April 2014.
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
CEN-CENELEC Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 12823-1:2014 ESIST EN 12823-1:2014
Examples of HPLC chromatograms . 13 Annex B (informative)
Precision data . 14 Annex C (informative)
Alternative HPLC systems . 15 Bibliography . 16
3 Principle Retinol is saponified by using methanolic or ethanolic potassium hydroxide solution and extracted by an appropriate solvent. The determination is carried out by high performance liquid chromatography (HPLC) with either fluorometric (F) or ultraviolet (UV) detection. The substances are identified on the basis of the retention times and determined by the external standard procedure using peak areas or heights, see [1] to [4]. 4 Reagents During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and water of at least grade 1 according to EN ISO 3696. 4.1 Methanol. 4.2 Ethanol absolute, volume fraction, m(C2H5OH) = 100 %. 4.3 Ethanol, m(C2H5OH) = 96 %. 4.4 Sodium sulfate, anhydrous. 4.5 KOH solution for saponification, in suitable mass concentrations, e.g. (KOH) = 50 g/100 ml or 60 g/100 ml, or alcoholic solutions, e.g. 28 g KOH in 100 ml of a mixture of 9 parts per volume of ethanol and 1 part per volume of water. 4.6 Antioxidants, such as ascorbic acid (AA), sodium ascorbate, sodium sulfide (Na2S), butylated hydroxytoluene (BHT), pyrogallol or hydroquinone. 4.7 Solvents and extraction solvents, such as diethyl ether (peroxide-free), di-isopropylether, light petroleum (boiling range of 40 °C to 60 °C), n-hexane, butanol, iso-octane or appropriate mixtures thereof. SIST EN 12823-1:2014
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The article discusses EN 12823-1:2014, a European Standard that outlines a method for measuring vitamin A levels in food using high performance liquid chromatography (HPLC). The method has been validated using samples of margarine and milk powder. The measurement of vitamin A is done by analyzing all-E-retinol, 13-Z-retinol, and β-carotene. This specific part of the standard focuses on the measurement of all-E-retinol and 13-Z-retinol. Additionally, the article mentions that the extract obtained after saponification can be used to determine β-carotene levels, but it is important to maintain a saponification temperature below 80 °C to prevent isomerisation and oxidation.
기사 제목: EN 12823-1:2014 - 식품 - HPLC에 의한 비타민 A 측정 - 제 1부: all-E-레티놀과 13-Z-레티놀의 측정 기사 내용: 이 유럽 표준은 HPLC(고성능 액체 크로마토그래피)를 사용하여 식품에서 비타민 A를 측정하는 방법을 명시하고 있다. 이 방법은 마가린과 분유를 포함한 샘플을 사용하여 모든 E-레티놀 농도가 653 µg/100 g에서 729 µg/100 g 사이이고, 13-Z-레티놀 농도가 30 µg/100 g에서 39 µg/100 g 사이인 교차 실험실 분석으로 검증되었다. 비타민 A 함량은 all-E-레티놀, 13-Z-레티놀 및 β-카로틴의 측정을 통해 수행된다. 이 부분은 all-E-레티놀과 13-Z-레티놀의 측정을 다룬다. 본 방법에서 사포니화 후 얻은 추출물은 EN 12823-2:2000 "식품 - HPLC에 의한 비타민 A의 측정 - 제 2부: β-카로틴의 측정"에 설명된 대로 β-카로틴의 측정에 사용될 수 있다. 이 경우, 사포니화 온도는 β-카로틴의 이성질화와 산화를 방지하기 위해 가능한 80°C를 넘지 않도록 주의해야 한다.
記事のタイトル：EN 12823-1:2014 - 食品 - 高性能液体クロマトグラフィーによるビタミンAの測定 - 第1部：all-Eレチノールと13-Zレチノールの測定 記事の内容：この欧州標準では、高性能液体クロマトグラフィー（HPLC）を使用して食品中のビタミンAを測定する方法が定められています。この方法は、マーガリンとミルクパウダーのサンプルを使用して検証されました。 all-Eレチノールの濃度は653 µg/100 gから729 µg/100 g、13-Zレチノールの濃度は30 µg/100 gから39 µg/100 gに及ぶものです。ビタミンAの含有量は、all-Eレチノール、13-Zレチノール、およびβ-カロチンの測定によって行われます。この部分では、all-Eレチノールと13-Zレチノールの測定が扱われます。 また、この方法で皂化後に得られた抽出物は、EN 12823 2:2000「食品 - 高性能液体クロマトグラフィーによるビタミンAの測定 - 第2部：β-カロチンの測定」で説明されているように、β-カロチンの測定に使用することができます。この場合、β-カロチンの異性化と酸化を防ぐために、皂化温度は80°C以下とする必要があります。
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