EN 14148:2003

Foodstuffs - Determination of vitamin K1 by HPLC

This draft European Standard specifies a method for the determination of vitamin K1 in foodstuffs by high performance liquid chromatography (HPLC). The determination of Vitamin K1 content is carried out by measurement of reduced phylloquinone. The method has been validated for milk and milk products, however laboratory experiences exist which show that the method is also applicable to other type of foodstuffs.

Lebensmittel - Bestimmung von Vitamin K1 mit HPLC

Diese Europäische Norm legt ein Verfahren zur Bestimmung von Vitamin K1 in Lebensmitteln mit Hochleistungsflüssigchromatographie (HPLC) fest. Der Gehalt an Vitamin K1 wird durch die Messung von reduziertem Phyllochinon bestimmt. Das Verfahren wurde für Milch und Milchprodukte validiert, es gibt jedoch experimentelle Labor-Erfahrungen, die zeigen, dass das Verfahren auch auf andere Lebensmittel anwendbar ist.

Produits alimentaires - Dosage de la vitamine K1 par CLHP

La présente Norme européenne spécifie une méthode de dosage de la vitamine K1 présente dans les produits alimentaires par chromatographie liquide haute performance (CLHP). La détermination de la teneur en vitamine K1 est effectuée par mesurage de la forme réduite de la phylloquinone. Cette méthode a été validée pour le lait et les produits laitiers pour nourrissons. Toutefois, des expériences en laboratoire ont montré que cette méthode est également applicable aux autres catégories de produits alimentaires [10].

Živila – Določevanje vitamina K1 s HPLC

General Information

• Status: Published
• Publication Date: 01-Jul-2003
• ICS: 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
• Completion Date: 17-Sep-2024
• Ref Project: SIST EN 14148:2003 - Foodstuffs - Determination of vitamin K1 by HPLC

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von Vitamin K1 mit HPLCProduits alimentaires - Dosage de la vitamine K1 par CLHPFoodstuffs - Determination of vitamin K1 by HPLC67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 14148:2003SIST EN 14148:2003en01-november-2003SIST EN 14148:2003SLOVENSKI
STANDARD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 14148July 2003ICS 67.050English versionFoodstuffs - Determination of vitamin K1 by HPLCProduits alimentaires - Dosage de la vitamine K1 par CLHPLebensmittel - Bestimmung von Vitamin K1 mit HPLCThis European Standard was approved by CEN on 2 May 2003.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the 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 translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,Hungary, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Slovakia, Spain, Sweden, Switzerland and UnitedKingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2003 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 14148:2003 ESIST EN 14148:2003

Figures.11Annex B (informative)
Precision data.13Annex C (informative)
Alternative HPLC-Systems.14Bibliography.15SIST EN 14148:2003

%4.2.6 Light petroleum, bp. 35 °C to 60 °C, p.a.4.2.7 Potassium hydroxide, w(KOH)
%4.2.8 Potassium hydroxide solution, substance concentration c(KOH) = 10 mol/lSIST EN 14148:2003

%4.2.15 Phosphate buffer pH 7,9 to 8,0Dissolve 54,0 g of potassium dihydrogen phosphate (4.2.9) in approximately 350 ml of water, adjust the pH to 7,9to 8,0 with potassium hydroxide solution (4.2.8) and dilute to 500 ml with water.4.2.16 Zinc chloride-acetate solutionWeigh 13,7 g of zinc chloride (4.2.13), 4,1 g of anhydrous sodium acetate (4.2.11) and 3,0 g of acetic acid (4.2.12)in a 50 ml volumetric flask, dissolve in methanol (4.2.1) and dilute to 50 ml with methanol.4.2.17 Lipase type VIIe.g. from Candida rugosa, activity ca. 1000 U/mg or suitable alternative1); other enzyme sources fromPseudomonas and Rhizopus species can also be used considering the different activity profile.4.2.18 HPLC Mobile phaseMix 100 ml of dichloromethane (4.2.4), 900 ml of methanol (4.2.1) and 5 ml of zinc chloride-acetate solution(4.2.16). Filter through a 0,45 mm filter.4.3 Vitamin K1 standard substance (Phyllochinone, 3-Phythylmenadione), w(C31H46O2) ³ 99%Vitamin K1 can be obtained from various suppliers. The purity of the phylloquinone standard may vary. It istherefore necessary to determine the concentration of the calibration solution by UV-spectrometry (seeconcentration test 4.4.4)4.4 Stock solutions4.4.1 PrecautionsVitamin K1 is very sensitive to light. Measures have to be taken to protect the standard and the correspondingsolutions during the whole procedure e.g. by using generally brown glass ware.4.4.2 Vitamin K1 stock solution I, mass concentration r(C31H46O2) » 1,0 mg/mlWeigh accurately approximately 100 mg of vitamin K1 standard substance (4.3) into a 100 ml volumetric flaskdissolve in methanol (4.2.1) and dilute to 100 ml. This solution can be stored under nitrogen for 3 months at -20 °Cin the dark.NOTEThe amount of vitamin K1 can be difficult to dissolve in methanol.
1)e.g. L-1754; Sigma Chemical Co, P.O. 14508, St. Louis, MO 63178 USA. This product was used in the interlaboratorystudy. This information is given for the convenience of users of this standard and does not constitute an endorsement by CEN ofthe product named. Equivalent products may be used if they can be shown to lead to the same results.SIST EN 14148:2003
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