Foodstuffs - Determination of vitamin E by high performance liquid chromatography - Measurement of α-, β-, γ- and δ-tocopherol

This European Standard specifies a method for the determination of vitamin E in foods by high performance liquid chromatography (HPLC). The determination of vitamin E content is carried out by measurement of α-, β , γ- and δ-tocopherol. This method has been validated in two interlaboratory studies. The first study was for the analysis of α-tocopherol in margarine and milk powder ranging from 9,89 mg/100 g to 24,09 mg/100 g. The second study was for the analysis of α-, β-, γ- and δ-tocopherol in milk powder and of α-, and β-tocopherol in oat powder ranging from 0,057 mg/100 g (β-tocopherol) to 10,2 mg/100 g (α-tocopherol).
NOTE   The vitamin E activity can be calculated from the tocopherol content assuming appropriate factors as given in [1], [2], [3] and [4].

Lebensmittel - Bestimmung von Vitamin E mit Hochleistungs-Flüssigchromatographie - Bestimmung von α-, β-, γ- und δ-Tocopherol

Diese Europäische Norm legt ein Verfahren zur Bestimmung von Vitamin E in Lebensmitteln mit Hochleistungs-Flüssigchromatographie (HPLC) fest. Die Bestimmung des Vitamin E-Gehalts erfolgt anhand der Bestimmung von -, -, - und -Tocopherol. Diese Bestimmung wurde in 2 Ringversuchen validiert. Im ersten Ringversuch wurde auf Tocopherol in Margarine und Milchpulver von 9,89 mg/100 g bis 24,09 mg/100 g untersucht. Im zweiten Ringversuch wurde -, -, - und -Tocopherol in Milchpulver und - und -Tocopherol in Haferpulver von 0,057 mg/100 g (-Tocopherol) bis 10,2 mg/100 g (-Tocopherol) untersucht.
Die Vitamin E-Aktivität kann aus den Tocopherolgehalten unter Verwendung geeigneter Faktoren, auf die in [1], [2], [3] und [4] verwiesen wird, berechnet werden.

Produits alimentaires - Détermination de la teneur en vitamine E par chromatographie liquide haute performance - Dosage des α-, β-, γ- et δ-tocophérols

La présente Norme européenne spécifie une méthode de dosage de la vitamine E présente dans les produits alimentaires par chromatographie liquide haute performance (CLHP). La détermination de la teneur en vitamine E est effectuée par dosage des α-, β-, γ- et δ-tocophérols. Cette méthode a été validée lors de deux essais interlaboratoires. Le premier essai concernait l’analyse de l’α-tocophérol dans la margarine et le lait en poudre, entre 9,89 mg/100 g et 24,09 mg/100 g. Le deuxième essai s’appliquait à l’analyse des α-, β-, γ- et δ-tocophérols dans le lait en poudre et des α- et β-tocophérols dans la poudre d’avoine, entre 0,057 mg/100 g (β-tocophérol) et 10,2 mg/100 g (α-tocophérol).
L’activité de la vitamine E peut être calculée à partir de la teneur en tocophérol, en utilisant les facteurs appropriés tels que mentionnés en [1], [2], [3] et [4].

Živila - Določevanje vitamina E s tekočinsko kromatografijo visoke ločljivosti - Merjenje α-, β-, γ- in δ-tokoferolov

Ta evropski standard določa metodo za določanje vitamina E v živilih s tekočinsko kromatografijo visoke ločljivosti (HPLC). Določanje vsebnosti vitamina E poteka z merjenjem α-, β-, γ- in δ-tokoferolov. Ta metoda je bila validirana z dvema medlaboratorijskima študijama. Prva študija je bila namenjena analizi α-tokoferolov v margarini in mleku v prahu, ki sta vsebovala od 9,89 mg/100 g do 24,09 mg/100 g α-tokoferolov. Druga študija je bila namenjena analizi α-, β-, γ- in δ-tokoferolov v mleku v prahu ter α- in β-tokoferolov v ovsu v prahu, ki sta vsebovala 0,057 mg/100 g β-tokoferolov do 10,2 mg/100 g α-tokoferolov. OPOMBA Aktivnost vitamina E je mogoče izračunati iz količine tokoferolov, če veljajo ustrezni dejavniki iz [1], [2], [3] in [4].

General Information

Status
Published
Publication Date
03-Jun-2014
ICS
67.040 - Food products in general
 
67.050 - General methods of tests and analysis for food products
Technical Committee
CEN/TC 275 - Food analysis - Horizontal methods
Drafting Committee
CEN/TC 275/WG 9 - Vitamins and Carotenoids
Current Stage
9093 - Decision to confirm - Review Enquiry
Start Date
28-Oct-2020
Completion Date
23-Sep-2025
Ref Project
SIST EN 12822:2014 - Foodstuffs - Determination of vitamin E by high performance liquid chromatography - Measurement of α-, β-, γ- and δ-tocopherol

Relations

Replaces
EN 12822:2000 - Foodstuffs - Determination of vitamin E by high performance liquid chromatography - Measurement of alpha-, beta-, gamma-, and delta-tocopherols
Effective Date
16-Jul-2011

Overview

EN 12822:2014 (CEN) is a European standard that specifies a validated method for the determination of vitamin E in foodstuffs by high performance liquid chromatography (HPLC). The method measures individual tocopherols - α‑, β‑, γ‑ and δ‑tocopherol - and has been validated in interlaboratory studies for common food matrices such as margarine, milk powder and oat powder. The standard supports reliable quantification of tocopherols to express vitamin E content and, where required, convert tocopherol mass to vitamin E activity using published factors.

Key topics and technical requirements

  • Analytes covered: α‑, β‑, γ‑ and δ‑tocopherol (individual tocopherol measurement by HPLC).
  • Analytical principle: HPLC separation with fluorescence detection (preferred - excitation 295 nm, emission 330 nm) or UV detection (e.g., 292 nm) using external (or validated internal) standards and peak-area quantification.
  • Sample preparation: Homogenization, protection from light, saponification and solvent extraction where needed; special procedures for oils, margarine and butter (fat isolation) and for complex matrices. Typical saponification times 15–40 min at 80–100 °C.
  • Chromatography systems: Normal phase (silica) and reversed phase (C18) columns are allowed; mobile phases and separation parameters are specified (examples: n‑hexane with modifiers for NP, methanol/water mixes for RP). Annex C describes alternative HPLC systems.
  • Standards and calibration: Use certified tocopherol standards; check stock solution concentration and purity by UV spectrometry. Store standards protected from light and below 4 °C.
  • Quality and precision: Method validated in two interlaboratory studies - α‑tocopherol in margarine and milk powder (≈9.89–24.09 mg/100 g) and multi‑tocopherol analyses including lower levels (down to 0.057 mg/100 g up to 10.2 mg/100 g). Annex B provides precision data.
  • Safety and good practice: Use analytical-grade reagents and protect samples from degradation (light, heat, oxygen); the standard includes a safety warning about hazardous materials and operations.

Applications and who uses it

  • Food testing and quality control laboratories performing vitamin E / tocopherol analysis.
  • Nutritional labeling and regulatory compliance testing for food manufacturers and inspection bodies.
  • Research laboratories analyzing lipid‑soluble antioxidants in dairy, margarine, cereals (e.g., oat powder), infant formula and other food matrices.
  • Method validation teams and proficiency testing organizers comparing laboratory performance.

Related standards

  • EN ISO 3696 - Water for analytical laboratory use (referenced for reagent water quality).
  • EN 12822 includes informative Annexes A (chromatograms), B (precision data) and C (alternative HPLC systems).
  • The standard refers to literature for conversion of tocopherol mass to vitamin E activity.

Keywords: EN 12822:2014, vitamin E determination, HPLC, tocopherol analysis, α‑tocopherol, food analysis, CEN standard, saponification, fluorescence detection.

Frequently Asked Questions

EN 12822:2014 is a standard published by the European Committee for Standardization (CEN). Its full title is "Foodstuffs - Determination of vitamin E by high performance liquid chromatography - Measurement of α-, β-, γ- and δ-tocopherol". This standard covers: This European Standard specifies a method for the determination of vitamin E in foods by high performance liquid chromatography (HPLC). The determination of vitamin E content is carried out by measurement of α-, β , γ- and δ-tocopherol. This method has been validated in two interlaboratory studies. The first study was for the analysis of α-tocopherol in margarine and milk powder ranging from 9,89 mg/100 g to 24,09 mg/100 g. The second study was for the analysis of α-, β-, γ- and δ-tocopherol in milk powder and of α-, and β-tocopherol in oat powder ranging from 0,057 mg/100 g (β-tocopherol) to 10,2 mg/100 g (α-tocopherol). NOTE The vitamin E activity can be calculated from the tocopherol content assuming appropriate factors as given in [1], [2], [3] and [4].

This European Standard specifies a method for the determination of vitamin E in foods by high performance liquid chromatography (HPLC). The determination of vitamin E content is carried out by measurement of α-, β , γ- and δ-tocopherol. This method has been validated in two interlaboratory studies. The first study was for the analysis of α-tocopherol in margarine and milk powder ranging from 9,89 mg/100 g to 24,09 mg/100 g. The second study was for the analysis of α-, β-, γ- and δ-tocopherol in milk powder and of α-, and β-tocopherol in oat powder ranging from 0,057 mg/100 g (β-tocopherol) to 10,2 mg/100 g (α-tocopherol). NOTE The vitamin E activity can be calculated from the tocopherol content assuming appropriate factors as given in [1], [2], [3] and [4].

EN 12822:2014 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 12822:2014 has the following relationships with other standards: It is inter standard links to EN 12822:2000. Understanding these relationships helps ensure you are using the most current and applicable version of the standard.

You can purchase EN 12822:2014 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 CEN standards.

Standards Content (Sample)


2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 12822:2014SIST EN 12822:2014en,fr,de01-september-2014SIST EN 12822:2014SLOVENSKI
STANDARDSIST EN 12822:20001DGRPHãþD

EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 12822
June 2014 ICS 67.050 Supersedes EN 12822:2000
English Version
Foodstuffs - Determination of vitamin E by high performance liquid chromatography - Measurement of $-, ß-, - and /-tocopherol
Produits alimentaires - Détermination de la teneur en vitamine E par chromatographie liquide haute performance - Dosage des $-, ß-, - et /-tocophérols
Lebensmittel - Bestimmung von Vitamin E mit Hochleistungs-Flüssigchromatographie - Bestimmung von $-, ß-, - und /-Tocopherol This European Standard was approved by CEN on 17 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 12822:2014 E SIST EN 12822:2014

Examples of HPLC chromatograms . 14 Annex B (informative)
Precision data . 16 Annex C (informative)
Alternative HPLC systems . 18 Bibliography . 19
1) This information is given for convenience of users of this European Standard and does not and does not constitute and endorsement by CEN. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 12822:2014

-tocopherol. If an UV-detector is used to monitor the chromatography, a more concentrated solution shall be used. The standard solution shall be stored protected from light and at a temperature below 4 °C and should be checked as described in 4.10.5. 4.11.2 ptandard solution of a mixture of .--- and i-tocopherol Pipette e.g. 10 ml of each of the stock solutions (4.10) into a one-mark 100 ml volumetric flask and dilute to the mark with the appropriate solvent (for NP e.g. n-hexane, for RP e.g. methanol). The standard solution should have a mass concentration of 1
2) Lichrosorb® Si 60, Spherisorb® Si, Hypersil® Si, Lichrospher® 100 DIOL, Spherisorb® ODS and Hypersil® ODS are examples of suitable products available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN. SIST EN 12822:2014
...

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제목: EN 12822:2014 - 식품 - 고성능 액체 크로마토그래피에 의한 비타민 E의 측정 - α-, β-, γ-, δ-토코페롤의 측정 내용: 이 유럽 표준은 고성능 액체 크로마토그래피(HPLC)를 통한 식품에서 비타민 E를 측정하기 위한 방법을 명시한다. 비타민 E 함량은 α-, β-, γ-, δ-토코페롤의 측정을 통해 수행된다. 이 방법은 두 개의 실험실 간 연구에서 검증되었다. 첫 번째 연구는 마가린과 분유에서 9.89 mg/100 g에서 24.09 mg/100 g까지의 α-토코페롤 분석을 위한 것이었다. 두 번째 연구는 분유에서 α-, β-, γ-, δ-토코페롤과 오트분말에서 α- 및 β-토코페롤의 분석을 위한 것이었다. 분석범위는 0.057 mg/100 g(β-토코페롤)에서 10.2 mg/100 g(α-토코페롤)까지였다. 참고: 적절한 요인 [1], [2], [3], [4]을 가정하여 토코페롤 함량으로부터 비타민 E의 활성도를 계산할 수 있다.

記事のタイトル：EN 12822:2014 - 食品 - 高性能液体クロマトグラフィーによるビタミンEの測定 - α-, β-, γ-, δ-トコフェロールの測定 記事の内容：この欧州標準は、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Eを測定する方法を規定しています。ビタミンEの含有量は、α-, β-, γ-, δ-トコフェロールの測定によって行われます。この方法は、2つの相互検証研究によって検証されました。第1の研究では、マーガリンとミルクパウダー中のα-トコフェロールの分析を9.89 mg/100 gから24.09 mg/100 gまでの範囲で行いました。第2の研究では、ミルクパウダー中のα-, β-, γ-, δ-トコフェロールおよびオートパウダー中のα-およびβ-トコフェロールの分析を0.057 mg/100 g（β-トコフェロール）から10.2 mg/100 g（α-トコフェロール）までの範囲で行いました。 注記：適切な係数[1]、[2]、[3]、[4]を前提として、トコフェロールの含有量からビタミンEの活性度を計算することができます。

The article discusses the European Standard EN 12822:2014, which provides a method for determining the vitamin E content in food using high performance liquid chromatography (HPLC). The method measures α-, β-, γ-, and δ-tocopherol. The validity of the method was confirmed through two interlaboratory studies. The first study focused on the analysis of α-tocopherol in margarine and milk powder, while the second study analyzed α-, β-, γ-, and δ-tocopherol in milk powder and α- and β-tocopherol in oat powder. The vitamin E activity can be calculated based on the tocopherol content, using appropriate factors provided in references 1, 2, 3, and 4.