ISO INTERNATIONAL STANDARD 23443 First edition 2020-07 Infant formula and adult nutritionals Determination of B-carotene, lycopene and lutein by reversed-phase ultra-high performance liquid chromatography (RP-UHPLC) Formules infantiles etproduits nutritionnels pouradultes- Determination du beta-carotene, du lycopene et de la luteine par chromatographie liquide ultra haute performance d phase inversee Reference number IS023443:2020(E) ISO @ IS0 2020 IS023443:2020(E) MhhmMni a a COPYRIGHTPROTECTEDDOCUMENT IS02020 All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may below or Iso's member body in the country of the requester. ISO copyright office CP 401: Ch. de Blandonnet 8 CH-1214 Vernier, Geneva Phone: +41 22 749 01 11 Email:copyright@iso.org Website: www.iso.org Published in Switzerland ii @ IS0 2020 - All rights reserved IS023443:2020(E) Contents Page Foreword iv Introduction. .V 1 Scope. .1 2 Normative references .1 3 Terms and definitions .1 4 Principle .2 5 Reagents and materials .2 5.1 Reagents. 12 5.2 Standards 3 5.3 Standards preparation 4 .6 6 Apparatus. 7 Procedure. .7 7.1 Samplepreparation 7 7.2 Chromatography 9 7.2.1 Chromatographic conditions 9 7.2.2 System suitability checks 9 Calculations. 8 .10 8.1 Determination of purity 10 8.1.1 General. 10 8.1.2 Spectrophotometric purity (Ps) 10 8.1.3 Chromatographic purity (Pc) 10 8.1.4 Reference standard purity (P) 11 8.2 Concentrationofeachcarotenoid in standard solutions. 11 8.2.1 Stocksolutionconcentrations. 11 8.2.2 Apocarotenalworking solution concentration 11 8.2.3 Apocarotenal intermediate solution concentration 11 8.2.4 Carotenoid concentrations inmixed carotenoid intermediate solution .12 .12 8.2.5 Concentrations ofcarotenoidanalytes in calibrations solutions. 8.2.6 Concentration ofapocarotenal internal standard in calibrations solutions 12 8.3 Calculate calibration curve 12 8.4 Mass of apocarotenal 13 8.5 Contents ofall-trans-β-carotene, cis isomers of β-carotene and total β-carotene 13 8.6 Contents of total lycopene 14 9 Precision .15 9.1 General. 15 9.2 Repeatability 15 9.3 Reproducibility .15 10 Test report. .15 AnnexA(informative)Examplechromatograms .17 AnnexB(informative)Precisiondata .22 AnnexC (informative)Determinationof lutein .25 Bibliography .32 @ IS0 2020 - All rights reserved iii