EN 14122:2014
(Main)Foodstuffs - Determination of vitamin B1 by high performance liquid chromatography
Foodstuffs - Determination of vitamin B1 by high performance liquid chromatography
This European Standard specifies a method for the determination of vitamin B1 in food by high performance liquid chromatography (HPLC) with enzymatic treatment and pre- or post-column derivatization. This method has been validated in two interlaboratory studies. The first study was for the analysis of samples of whole meal flour, milk powder/spray dried milk, freeze-dried mixed vegetables and freeze-dried pig's liver ranging from 0,295 mg/100 g to 0,807 mg/100 g. The second study was for the analysis of samples of tube feeding solution, baby food with vegetables, powdered milk, meal with fruits, yeast, cereal, chocolate powder and food supplement ranging from 0,11 mg/100 g to 486 mg/100 g. Vitamin B1 is the mass fraction of total thiamin including its phosphorylated derivatives.
For further information on the validation, see Clause 8 and Annex B.
Lebensmittel - Bestimmung von Vitamin B1 mit Hochleistungs-Flüssigchromatographie
Diese Europäische Norm legt ein Verfahren zur Bestimmung von Vitamin B1 in Lebensmitteln mit Hochleistungsflüssigchromatographie (HPLC) mit Enzymbehandlung und Vor- oder Nachsäulenderivati¬sierung fest. Diese Bestimmung wurde in zwei Ringversuchen validiert. In dem einen Ringversuch wurde Vollkornmehl, Milchpulver/sprühgetrocknete Milch, gefriergetrocknetes Mischgemüse und gefrier¬getrocknete Schweineleber mit Gehalten von 0,295 mg/100 g bis 0,807 mg/100 g analysiert. Im zweiten Ringversuch wurde Krankenhausnahrung, Babynahrung mit Gemüse, Milchpulver, Fertignahrungsmittel mit Früchten, Hefe, Getreide, Schokoladenpulver und Nahrungsergänzungsmittel von 0,11 mg/100 g bis 468 mg/100 g untersucht. Vitamin B1 ist der Massenanteil an Gesamtthiamin einschließlich der phosphorylierten Derivate.
Produits alimentaires - Détermination de la teneur en vitamine B1 par chromatographie liquide haute performance
La présente Norme européenne spécifie une méthode de dosage de la vitamine B1 présente dans les produits alimentaires par chromatographie liquide haute performance (CLHP) avec traitement enzymatique et dérivation pré- ou post-colonne. Cette méthode a été validée lors de deux essais interlaboratoires. Le premier essai concernait l’analyse d’échantillons de farine complète, de lait en poudre/lait séché par atomisation, d’assortiments de légumes lyophilisés et de foie de porc lyophilisé allant de 0,295 mg/100 g à 0,807 mg/100 g. Le deuxième essai s’appliquait à l’analyse d’échantillons de solution pour alimentation entérale, d’aliment pour bébé aux légumes, de lait en poudre, de substitut de repas aux fruits, de levure, de céréale, de chocolat en poudre et de complément alimentaire allant de 0,11 mg/100 g à 486 mg/100 g. La vitamine B1 correspond à la fraction massique de thiamine totale, y compris ses dérivés phosphorylés.
Pour plus d'informations sur la validation, voir l'Article 8 et l'Annexe B.
Živila - Določevanje vitamina B1 s tekočinsko kromatografijo visoke ločljivosti
Standard EN 14122 navaja metodo za določanje vitamina B1 v živilih s tekočinsko kromatografijo visoke ločljivosti (HPLC) z encimsko obdelavo ter pred- ali post-kolonsko derivatizacijo. Ta metoda je bila validirana z dvema medlaboratorijskima študijama. Prva študija je bila namenjena analizi vzorcev polnozrnate moke, mleka v prahu/atomiziranega mleka, zmrznjene mešane zelenjave in zmrznjenih prašičjih jeter, ki vsebujejo od 0,295 mg/100 g do 0,807 mg/100 g vitamina B1. Druga študija je bila namenjena analizi vzorcev raztopin za krmljenje, zelenjavnih formul za dojenčke, mleka v prahu, grobe moke s sadjem, kvasa, žitaric, čokolade v prahu in prehranskih nadomestil, ki vsebujejo od 0,11 mg/100 g do 486 mg/100 g vitamina B1. Vitamin B1 je masni delež celotne vrednosti tiamina, vključno z njegovimi fosforiliranimi derivati. Za več informacij glede validacije glejte točko 8 in dodatek B.
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2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von Vitamin B1 mit Hochleistungs-FlüssigchromatographieProduits alimentaires - Dosage de la vitamine B1 par chromatographie liquide haute performanceFoodstuffs - Determination of vitamin B1 by high performance liquid chromatography67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 14122:2014SIST EN 14122:2014en,fr,de01-september-2014SIST EN 14122:2014SLOVENSKI
STANDARDSIST EN 14122:2003/AC:2006SIST EN 14122:20031DGRPHãþD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 14122
June 2014 ICS 67.050 Supersedes EN 14122:2003English Version
Foodstuffs - Determination of vitamin B1 by high performance liquid chromatography
Produits alimentaires - Détermination de la teneur en vitamine B1 par chromatographie liquide haute performance Lebensmittel - Bestimmung von Vitamin B1 mit Hochleistungs-Flüssigchromatographie 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.
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Examples of HPLC chromatograms . 13 Annex B (informative)
Precision data . 15 Annex C (informative)
Alternative HPLC systems . 18 Annex D (informative)
Vitamin B1 compound: 2-(1-hydroxyethyl)thiamin (HET) performing post-column derivatization . 19 Bibliography . 21
1) The information of the suppliers of Taka-Diastase, Pfaltz & Bauer, Waterbury, CT 06708, USA (No T00040), and Serva is given for the convenience of users of this European standard and does not constitute an endorsement by CEN of the product named. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 14122:2014
HCl) ≈ 0,1 mg/ml. Dissolve an accurately weighed amount of the thiamin chloride hydrochloride standard substance (4.23.1) in a defined volume of an appropriate solvent, for example 10 mg of vitamin B1 standard substance in 100 ml of hydrochloric acid solution (4.6). This solution can be stored for four weeks at + 4 °C. 4.24.2 Thiamin monophosphate stock solution, (C12H17ClN4O4PS) ≈ 0,1 mg/ml. Dissolve an accurately weighed amount of the thiamin monophosphate chloride (4.23.2) in a defined volume of an appropriate solvent, for example 10 mg of thiamin monophosphate chloride in 100 ml of hydrochloric acid solution (4.6). This solution can be stored for four weeks at - 20 °C. 4.24.3 Thiamin pyrophosphate stock solution, (C12H19ClN4O7P2S) ≈ 0,1 mg/ml. Dissolve an accurately weighed amount of the thiamin pyrophosphate chloride (4.23.3) in a defined volume of an appropriate solvent, for example 10 mg of the thiamin pyrophosphate chloride in 100 ml of hydrochloric acid solution (4.6). 4.24.4 Concentration tests - thiamin chloride hydrochloride Dilute 10 ml of the thiamin chloride hydrochloride solution (4.24.1) with hydrochloric acid solution (4.6) in a 100 ml volumetric flask to the mark. Measure the absorbance of this solution at the maximum of about 247 nm (A247) in a 1 cm cell against hydrochloric solution (4.6) in the reference cell using an UV spectrometer (5.1). Calculate the mass concentration, , in microgram per millilitre thiamin chloride hydrochloride solution (4.24.1) using Formula (1): ερ0001247⋅⋅=MA (1) where 0 is the molar absorption coefficient of thiamin chloride hydrochloride at the maximum wavelength of about 247 nm. The value is 14 200 l · mol−1 · cm−1. This value is calculated from the extinction coefficient, 1%1cmE = 421, in 0,1 mol/l HCl [8][7] and the molar mass, M = 337,21. The value is given with four significant figures; M is the molar mass, in grams per mol. The value is 337,21; A247 is the absorption value of the thiamin chloride hydrochloride solution. 4.25 Standard solutions 4.25.1 Thiamin chloride hydrochloride standard solution, (C12H17ClN4lS
HCl) ≈ 1
Pipette 1 ml to 10 ml of the thiamin chloride hydrochloride solution (4.24.1) into a 100 ml volumetric flask and dilute to the mark with the appropriate solvent, e.g. hydrochloric acid solution (4.6). This solution can be stored at 4 °C in the dark for 1 month. 4.25.2 Thiamin monophosphate standard solution, (C12H17ClN4O4mS) ≈ 1
Pipette 1 ml to 10 ml of the thiamin monophosphate solution (4.24.2) into a 100 ml volumetric flask and dilute to the mark with the appropriate solvent, e.g. hydrochloric acid solution (4.6). This solution can be stored at 4 °C in the dark for 1 month. SIST EN 14122:2014
Pipette 1 ml to 10 ml of the thiamin pyrophosphate solution (4.24.3) into a 100 ml volumetric flask and dilute to the mark with the appropriate solvent, e.g. hydrochloric acid solution (4.6). This solution can be stored at 4 °C in the dark for 1 month. 5 Apparatus Usual laboratory apparatus, glassware, and the following: 5.1 UV spectrometer, UV spectrometer, capable of measuring absorption at defined wavelengths (247 nm), with appropriate cells, e.g. of 1 cm length. 5.2 Autoclave or heating device, autoclave for extraction purpose, e.g. pressure cooker type, with pressu
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