SIST EN 15607:2009

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von d-Biotin mit HPLCProduits alimentaires - Dosage de la d-biotine par CLHPFoodstuffs - Determination of d-biotin 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 15607:2009SIST EN 15607:2009en,fr,de01-september-2009SIST EN 15607:2009SLOVENSKI
STANDARD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 15607May 2009ICS 67.050 English VersionFoodstuffs - Determination of d-biotin by HPLCProduits alimentaires - Dosage de la d-biotine par CLHPLebensmittel - Bestimmung von d-Biotin mit HPLCThis European Standard was approved by CEN on 23 April 2009.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 CEN 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 CEN Management Centre has the same status as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, 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 STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre:
Avenue Marnix 17,
B-1000 Brussels© 2009 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 15607:2009: ESIST EN 15607:2009

Typical chromatogram . 11Annex B (informative)
Precision data . 13Bibliography . 14 SIST EN 15607:2009

1 Katal (symbol ''kat'') is a derived SI unit of enzyme activity. One katal is that catalytic activity which will raise the rate of reaction by one mol/s in a specified assay system. SIST EN 15607:2009

has been used 2. 4.2.19 Avidin fluoresceine isothiocyanate (Avidin-FITC), labelled, 80 % protein, 2 mol to 4 mol FITC per mol of avidin 4.2.20 Stock solution reagent for post-column binding reaction, (avidin-FITC) = 50 mg/ml Dissolve 2,5 mg of avidin-FITC (4.2.19) in 50 ml of phosphate buffer solution pH = 7,0 (4.2.16). This solution is stable for 2 weeks at 4 °C. 4.2.21 Reagent for post-column binding reaction, (avidin-FITC) = 2 mg/ml Add 600 ml of phosphate buffer solution pH = 7,0 (4.2.16) to 25 ml of the stock solution (4.2.20). Filter this solution through a 0,45 µm filter (5.5). This solution is stable for 8 hours, screened from light. 4.2.22 HPLC mobile phase Mix 80 parts per volume of phosphate buffer solution pH = 6,0 (4.2.15) with 20 parts per volume of methanol (4.2.1). Filter this solution through a 0,45 µm filter (5.5). 4.2.23 Taka-diastase from Aspergillus Oryzae, enzyme activity is 1 500 nkat/mg (1 500 nkat/mg corresponds to 100 U/mg), suitable for samples with a high starch content. 4.3 Standard substances 4.3.1 General D-biotin and d-biocytin can be obtained from various suppliers. The baseline separation of d-biotin and d-biocytin shall be verified. So it is necessary to prepare a standard solution. The biotin content of the standard can be confirmed according to the European Pharmacopoeia procedure [6]. 4.3.2 d-biotin, w(C10H16N2O3S) ≥ 99 % 4.3.3 d-biocytin, w(C16H28N4O4S) ≥ 98 % 4.4 Stock solutions 4.4.1 d-biotin, (C10H16N2O3S)
= 100 µg/ml Dissolve an amount of the d-biotin standard substance (4.3.2), approximately 10 mg to the nearest 0,1 mg in 100 ml of distilled water. It may take 4 h to 5 h to dissolve. This solution is stable for 2 months at –18 °C.
2 This information 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 t
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