ISO INTERNATIONAL STANDARD 9167 First edition 2019-05 Rapeseed and rapeseed meals - Determination of glucosinolates content Method using high- performance liquid chromatography Graines et tourteaux de colza - Dosage des glucosinolates - Méthode par chromatographie liquide a haute performance Reference number IS0 9167:2019(E) so IS02019 IS0 9167:2019(E) COPYRIGHTPROTECTEDDOCUMENT @ ISO 2019 All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permission. Permission can be requested from either IsO at the address 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: +4122 749 0111 Fax: +41 22 749 09 47 Email: [email protected] Website: www.iso.org Published in Switzerland ii @ IS0 2019 - All rights reserved IS0 9167:2019(E) Contents Page Foreword ..iv Introduction. ..V 1 Scope. ..1 2 Normative references 3 Terms and definitions 4 Principle ..2 5 Reagents. .2 6 Apparatus ..4 7 Sampling. .5 8 Preparation of the test sample ..6 9 Procedure .6 9.1 Testportion .6 9.2 Extraction of glucosinolates 6 9.3 .1 Blank test 9.4 Preparation of ion-exchange columns 7 9.5 Purification and desulfatation. ..7 9.6 Chromatography with gradient elution .7 9.6.1 General 7 9.6.2 Adjustment of the apparatus.. ..8 10 Expression of results ..9 10.1 Calculation of the content of each glucosinolate .9 10.2 Relative proportionality factors ..9 10.3 Calculation of the total glucosinolate content. ..10 11 Precision ..10 11.1 Interlaboratory test .10 11.2 Repeatability .10 11.3 Reproducibility 10 12 Test report ..11 Annex A (informative) Results of interlaboratory trials - Gradient elution HPLC method ..12 Annex B (normative) Checking of the titre of the prepared internal standard solution. ..14 Annex C (normative) Preparation and test of purified sulfatase solution and checking of the desulphation step on ion-exchange columns. .15 Annex D (informative) HPLC system and column performance criteria qualification ..20 Annex E (informative) Elution in the isocratic mode ..22 Bibliography .28 @ IS0 2019 - All rights reserved ii