{
  "url": "https://openlegislation.uk/eu-regulation-2009-152/annex-VI/division-2/division-2.1/division-2.1.3/division-2.1.3.5",
  "json_url": "https://openlegislation.uk/eu-regulation-2009-152/annex-VI/division-2/division-2.1/division-2.1.3/division-2.1.3.5.json",
  "legislation": "eur/2009/152",
  "legislation_title": "Commission Regulation (EC) No 152/2009 of 27 January 2009 laying down the methods of sampling and analysis for the official control of feed (Text with EEA relevance)",
  "legislation_url": "https://openlegislation.uk/eu-regulation-2009-152",
  "path": "annex/VI/division/2/division/2.1/division/2.1.3/division/2.1.3.5",
  "kind": "division",
  "number": "2.1.3.5",
  "heading": "Use of staining reagents",
  "text": "In order to facilitate the correct identification of the constituents of animal origin, the operator may use staining reagents during the sample preparation in accordance with guidelines issued by the EURL-AP and published on its website.\nIn case Alizarin Red solution is used to colour the sediment, the following protocol shall apply:\nthe dried sediment shall be transferred into a glass test tube and rinsed twice with approximately 5 ml of ethanol (each time a vortex of 30 s shall be used, the solvent shall be let settle about 1 min 30 s and poured off).\nthe sediment shall be bleached by adding at least 1 ml sodium hypochlorite solution. The reaction shall be allowed to continue for 10 min. The tube shall be filled with water, the sediment shall be let settle 2-3 min, and the water and the suspended particles shall be poured off gently.\nthe sediment shall be rinsed twice more with about 10 ml of water (a vortex shall be used for 30 s, let settle, and pour off the water each time).\n2 to 10 drops of the Alizarin Red solution shall be added and the mixture shall be vortexed. The reaction shall be let occur for 30 s and the coloured sediment shall be rinsed twice with approximately 5 ml ethanol followed by one rinse with acetone (each time a vortex of 30 s shall be used, the solvent shall be let settle about 1 min and poured off).\nthe coloured sediment shall be dried.",
  "extent": "E+W+S+N.I.",
  "status": null,
  "version": "current",
  "annotations": [
    {
      "type": "F",
      "label": "F1",
      "text": "Substituted by Commission Regulation (EU) No 51/2013 of 16 January 2013 amending Regulation (EC) No 152/2009 as regards the methods of analysis for the determination of constituents of animal origin for the official control of feed (Text with EEA relevance)."
    }
  ],
  "outstanding_effects": [],
  "provenance": {
    "source": "legislation.gov.uk",
    "source_url": "https://www.legislation.gov.uk/eur/2009/152/annex/VI/division/2/division/2.1/division/2.1.3/division/2.1.3.5",
    "retrieved_at": "2026-11-01T14:51:38.610873+00:00"
  },
  "licence": "Open Government Licence v3.0",
  "licence_url": "https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/",
  "attribution": "Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives."
}