# L_2023068EN.01003701.xml

| 6.3.2023 | EN | Official Journal of the European Union | L 68/37 |
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6.3.2023

EN

Official Journal of the European Union

L 68/37

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COMMISSION REGULATION (EU) 2023/464

of 3 March 2023

amending, for the purpose of its adaptation to technical progress, the Annex to Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(Text with EEA relevance)

THE EUROPEAN COMMISSION,

Having regard to the Treaty on the Functioning of the European Union,

Having regard to Regulation (EC) No 1907/2006 of the European Parliament and of the Council of 18 December 2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), establishing a European Chemicals Agency, amending Directive 1999/45/EC and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC and Commission Directives 91/155/EEC, 93/67/EEC, 93/105/EC and 2000/21/EC (1), and in particular Article 13(2) and (3) thereof,

Whereas:

| (1) | Article 13(3) of Regulation (EC) No 1907/2006 provides that, where tests on substances are required to generate information on intrinsic properties of substances, they are to be conducted in accordance with the test methods laid down in a Commission Regulation or in accordance with other international test methods recognised as being appropriate by the Commission or the European Chemicals Agency. |
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(1)

Article 13(3) of Regulation (EC) No 1907/2006 provides that, where tests on substances are required to generate information on intrinsic properties of substances, they are to be conducted in accordance with the test methods laid down in a Commission Regulation or in accordance with other international test methods recognised as being appropriate by the Commission or the European Chemicals Agency.

| (2) | Commission Regulation (EC) No 440/2008 (2) contains, in its Annex, test methods recognised as being appropriate for generating information on the physicochemical, toxicological and eco-toxicological properties of chemicals for the purpose of Regulation (EC) No 1907/2006. |
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(2)

Commission Regulation (EC) No 440/2008 (2) contains, in its Annex, test methods recognised as being appropriate for generating information on the physicochemical, toxicological and eco-toxicological properties of chemicals for the purpose of Regulation (EC) No 1907/2006.

| (3) | Most of the test methods contained in the Annex to Regulation (EC) No 440/2008 are equivalent to internationally agreed and accepted methods (such as test guidelines of the Organisation for Economic Cooperation and Development). These methods are frequently reviewed and modified to reflect the state of science. |
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(3)

Most of the test methods contained in the Annex to Regulation (EC) No 440/2008 are equivalent to internationally agreed and accepted methods (such as test guidelines of the Organisation for Economic Cooperation and Development). These methods are frequently reviewed and modified to reflect the state of science.

| (4) | Repeating the full description of those internationally agreed and accepted methods in the Annex to Regulation (EC) No 440/2008 for the purpose of their incorporation into the Union legislation has led to delays in adapting that Regulation to scientific progress. As a consequence, the test methods laid down in the Annex to Regulation (EC) No 440/2008 are frequently not aligned with the most up-to-date version of the corresponding international methods. Relatedly, new international test methods are added to Regulation (EC) No 440/2008 only after a prolonged period of time. |
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(4)

Repeating the full description of those internationally agreed and accepted methods in the Annex to Regulation (EC) No 440/2008 for the purpose of their incorporation into the Union legislation has led to delays in adapting that Regulation to scientific progress. As a consequence, the test methods laid down in the Annex to Regulation (EC) No 440/2008 are frequently not aligned with the most up-to-date version of the corresponding international methods. Relatedly, new international test methods are added to Regulation (EC) No 440/2008 only after a prolonged period of time.

| (5) | This situation has created uncertainty for registrants under Regulation (EC) No 1907/2006, as well as for duty holders under other Union legislation, as to which methods should be used for the generation of data for the purposes of that Regulation and other legislation. Article 13(2) of Regulation (EC) No 1907/2006 provides that methods are to be regularly reviewed and improved with a view to reducing testing on vertebrate animals and the number of animals involved, and that the Commission is to, if appropriate, make a proposal as soon as possible to amend Regulation (EC) No 440/2008, so as to replace, reduce or refine animal testing. Furthermore, Article 13 of Directive 2010/63/EU of the European Parliament and of the Council (3) on the protection of animals used for scientific purposes, makes it a legal obligation in the Union to use an alternative method that does not entail the use of a live animal, instead of an animal method, once such method is recognised under the legislation of the Union. Any delays to the process of introducing new alternative methods into Regulation (EC) No 440/2008 therefore could hinder a timely uptake of such methods once they are adopted at international level. |
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(5)

This situation has created uncertainty for registrants under Regulation (EC) No 1907/2006, as well as for duty holders under other Union legislation, as to which methods should be used for the generation of data for the purposes of that Regulation and other legislation. Article 13(2) of Regulation (EC) No 1907/2006 provides that methods are to be regularly reviewed and improved with a view to reducing testing on vertebrate animals and the number of animals involved, and that the Commission is to, if appropriate, make a proposal as soon as possible to amend Regulation (EC) No 440/2008, so as to replace, reduce or refine animal testing. Furthermore, Article 13 of Directive 2010/63/EU of the European Parliament and of the Council (3) on the protection of animals used for scientific purposes, makes it a legal obligation in the Union to use an alternative method that does not entail the use of a live animal, instead of an animal method, once such method is recognised under the legislation of the Union. Any delays to the process of introducing new alternative methods into Regulation (EC) No 440/2008 therefore could hinder a timely uptake of such methods once they are adopted at international level.

| (6) | In the Decision in case 23/2018/SRS, the European Ombudsman suggested to the Commission to intensify its efforts to simplify and speed up the process for introducing new alternative test methods under Regulation (EC) No 440/2008. Also, the European Parliament, in its resolution 2021/2784(RSP) of 16 September 2021 on plans and actions to accelerate the transition to innovation without the use of animals in research, regulatory testing and education, recalled that Article 13 of Regulation (EC) No 1907/2006 requires that the test method requirements be updated as soon as non-animal methods become available. |
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(6)

In the Decision in case 23/2018/SRS, the European Ombudsman suggested to the Commission to intensify its efforts to simplify and speed up the process for introducing new alternative test methods under Regulation (EC) No 440/2008. Also, the European Parliament, in its resolution 2021/2784(RSP) of 16 September 2021 on plans and actions to accelerate the transition to innovation without the use of animals in research, regulatory testing and education, recalled that Article 13 of Regulation (EC) No 1907/2006 requires that the test method requirements be updated as soon as non-animal methods become available.

| (7) | Therefore, in order to ensure that Regulation (EC) No 440/2008 provides correct, up-to-date and relevant test methods that are appropriate to generate information under Regulation (EC) No 1907/2006, a table should be included in the Annex to that Regulation, giving a comprehensive list of such methods with a reference to the respective international test method. The inclusion of a reference to an international test method in the table should be considered as a recognition by the Commission of such method for the purposes of Article 13(3) of Regulation (EC) No 1907/2006. |
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(7)

Therefore, in order to ensure that Regulation (EC) No 440/2008 provides correct, up-to-date and relevant test methods that are appropriate to generate information under Regulation (EC) No 1907/2006, a table should be included in the Annex to that Regulation, giving a comprehensive list of such methods with a reference to the respective international test method. The inclusion of a reference to an international test method in the table should be considered as a recognition by the Commission of such method for the purposes of Article 13(3) of Regulation (EC) No 1907/2006.

| (8) | Full descriptions of test methods in Parts A, B and C of the Annex to Regulation (EC) No 440/2008 that no longer correspond to the most recent version of an international test method should be deleted from that Annex to prevent tests being performed following protocols that do not provide state-of-the-art scientific information. |
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(8)

Full descriptions of test methods in Parts A, B and C of the Annex to Regulation (EC) No 440/2008 that no longer correspond to the most recent version of an international test method should be deleted from that Annex to prevent tests being performed following protocols that do not provide state-of-the-art scientific information.

| (9) | Certain test methods set out in the Annex to Regulation (EC) No 440/2008, as well as their corresponding international test methods, are no longer considered appropriate to generate new information under Regulation (EC) No 1907/2006. The test methods B.22. Rodent dominant lethal test; B.25. Mouse heritable translocation; B.34. One-generation reproductive toxicity study; B.35. Two-generation reproductive toxicity study; B.39. Unscheduled DNA synthesis (UDS) test with mammalian liver cells in vivo; and C.15. Fish, Short term toxicity test on Embryo and Sac Fry Stages; should therefore be deleted from that Annex and there should be no entry referring to them in the table set out in that Annex. |
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(9)

Certain test methods set out in the Annex to Regulation (EC) No 440/2008, as well as their corresponding international test methods, are no longer considered appropriate to generate new information under Regulation (EC) No 1907/2006. The test methods B.22. Rodent dominant lethal test; B.25. Mouse heritable translocation; B.34. One-generation reproductive toxicity study; B.35. Two-generation reproductive toxicity study; B.39. Unscheduled DNA synthesis (UDS) test with mammalian liver cells in vivo; and C.15. Fish, Short term toxicity test on Embryo and Sac Fry Stages; should therefore be deleted from that Annex and there should be no entry referring to them in the table set out in that Annex.

| (10) | Regulation (EC) No 440/2008 should therefore be amended accordingly. |
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(10)

Regulation (EC) No 440/2008 should therefore be amended accordingly.

| (11) | The measures provided for in this Regulation are in accordance with the opinion of the Committee established under Article 133 of Regulation (EC) No 1907/2006, |
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(11)

The measures provided for in this Regulation are in accordance with the opinion of the Committee established under Article 133 of Regulation (EC) No 1907/2006,

HAS ADOPTED THIS REGULATION:

Article 1

The Annex to Regulation (EC) No 440/2008 is amended in accordance with the Annex to this Regulation.

Article 2

This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 3 March 2023.

For the Commission

The President

Ursula VON DER LEYEN

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(1)  
  OJ L 396, 30.12.2006, p. 1.

1

(2)  Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) (OJ L 142, 31.5.2008, p. 1).

2

(3)  Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes (OJ L 276, 20.10.2010, p. 33).

3

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ANNEX

The Annex to Regulation (EC) No 440/2008 is amended as follows:

| (1) | before Part A, the following Part 0 is inserted

  ‘
  PART 0


  INTERNATIONAL TEST METHODS RECOGNISED AS BEING APPROPRIATE FOR GENERATING INFORMATION ON INTRINSIC PROPERTIES OF SUBSTANCES FOR THE PURPOSES OF REGULATION (EC) No 1907/2006

  TABLE 1: TEST METHODS FOR PHYSICOCHEMICAL PROPERTIES OF THE SUBSTANCE







  Endpoint | Test method | Corresponding chapter, containing the full description of the test method, in Part A of this Annex (numbers in brackets indicate that the full description of the test method has been deleted from Part A; empty cell: no corresponding test method in Part A of this Annex) |
| --- | --- | --- | --- |
| Melting point/freezing point | OECD Test Guideline 102: Melting Point/Melting Range (1995) | A.1. |
| Boiling point | OECD Test Guideline 103: Boiling point (1995) | A.2. |
| Density | OECD Test Guideline 109: Density of Liquids and Solids (2012) | (A.3.) |
| Vapour pressure | OECD Test Guideline 104: Vapour Pressure (2006) | (A.4) |
| Surface tension | OECD Test Guideline 115: Surface Tension of Aqueous Solutions (1995) | A.5. |
| Water solubility | OECD Test Guideline 105: Water Solubility (1995) | A.6. |
| Partition coefficient n-octanol/water | OECD Test Guideline 107: Partition Coefficient (n-octanol/water): Shake-Flask Method (1995) | (A.8.) |
| OECD Test Guideline 123: Partition Coefficient (1-Octanol/Water): Slow-Stirring Method (2022) | A.23. |
| OECD Test Guideline 117: Partition Coefficient (n-octanol/water): HPLC Method (2022) | A.24. |
| Dissociation constant | OECD Test Guideline 112: Dissociation Constants in Water. (1981) | A.25. |
| Viscosity | OECD Test Guideline 114: Viscosity of Liquids (2012) | |
| Flash point | Test methods according to table 2.6.3 of Annex I, Part 2 of Regulation (EC) No 1272/2008 | |
| Lower and upper explosion limit | EN 1839:2017 – Determination of the explosion limits and the limiting oxygen concentration (LOC) for flammable gases and vapours | |
| Flammability | Test methods according to section 2.2.4.1. of Annex I, Part 2 of Regulation (EC) No 1272/2008 | |
| Test L.2: sustained combustibility test, Part III, section 32 of the UN RTDG Manual of Tests and Criteria | |
| Test N.1: test method for readily combustible solids, Part III, sub-section 33.2.4 of the UN RTDG Manual of Tests and Criteria | |
| Test N.5: test method for substances which in contact with water emit flammable gases, Part III, sub-section 33.5.4 of the UN RTDG Manual of Tests and Criteria | |
| Self-ignition temperature (solids) | Test N.4: test method for self-heating substances, Part III, sub-section 33.4.6 of the UN RTDG Manual of Tests and Criteria | |
| EN 15188:2020 – Determination of the spontanous ignition behaviour of dust accumulations | |
| Auto-ignition temperature (liquids, gases) | ISO/IEC 80079-20-1:2017 – Explosive atmospheres – Part 20-1: Material characteristics for gas and vapour classification – Test methods and data | |
| Decomposition temperature | Test Series H, part II, section 28, of the UN RTDG Manual of Tests and Criteria | |
| Explosive properties | Test methods according to Test series 1-3, Part I, sections 11-13 of the UN RTDG Manual of Tests and Criteria | |
| EU Test method A.14 Explosive Properties | A.14 |
| Oxidising properties | Test method according to section 2.4.4. of Annex I, Part 2 of Regulation (EC) No 1272/2008 | |
| Test O.2: test for oxidizing liquids, Part III, sub-section 34.4.2 of the UN RTDG Manual of Tests and Criteria | |
| Test O.1: Test for oxidizing solids, Part III, sub-section 34.4.1 of the UN RTDG Manual of Tests and Criteria | |
| Test O.3 Gravimetric test for oxidizing solids, Part III, sub-section 34.4 3 of the UN RTDG Manual of Tests and Criteria | |
| Pyrophoricity | Test N.3: test method for pyrophoric liquids, Part III, sub-section 33.3.1.5 of the UN RTDG Manual of Tests and Criteria | |
| Test N.2: test method for pyrophoric solids, Part III, sub-section 33.3.1.4 of the UN RTDG Manual of Tests and Criteria | |
| Granulometry/particle characteristics | EU test method A.22. Length Weighted Geometric Mean Diameter of Fibres | A.22. |
| ISO 13318 – Determination of Particle Size Distribution by Centrifugal Liquid Sedimentation Methods | |
| ISO 21501 – Determination of Particle Size Distribution – Single Particle Light Interaction Methods | |
| OECD Test Guideline 124: Determination of the Volume Specific Surface Area of Manufactured Nanomaterials (2022) | |
| OECD Test Guideline 125: Particle Size and Particle Size Distribution of Nanomaterials (2022) | |
| pH | OECD Test Guideline 122: Determination of pH, Acidity and Alkalinity (2013) | |
| Properties of polymers | OECD Test Guideline 118: Determination of the Number-Average Molecular Weight and the Molecular Weight Distribution of Polymers using Gel Permeation Chromatography (1996) | A.18. |
| OECD Test Guideline 119: Determination of the Low Molecular Weight Content of a Polymer Using Gel Permeation Chromatography (1996) | A.19. |
| OECD Test Guideline 120: Solution/Extraction Behaviour of Polymers in Water (2000) | (A.20.) |

(1)

before Part A, the following Part 0 is inserted

‘
  PART 0

INTERNATIONAL TEST METHODS RECOGNISED AS BEING APPROPRIATE FOR GENERATING INFORMATION ON INTRINSIC PROPERTIES OF SUBSTANCES FOR THE PURPOSES OF REGULATION (EC) No 1907/2006

TABLE 1: TEST METHODS FOR PHYSICOCHEMICAL PROPERTIES OF THE SUBSTANCE

Endpoint

Test method

Corresponding chapter, containing the full description of the test method, in Part A of this Annex (numbers in brackets indicate that the full description of the test method has been deleted from Part A; empty cell: no corresponding test method in Part A of this Annex)

Melting point/freezing point

OECD Test Guideline 102: Melting Point/Melting Range (1995)

A.1.

Boiling point

OECD Test Guideline 103: Boiling point (1995)

A.2.

Density

OECD Test Guideline 109: Density of Liquids and Solids (2012)

(A.3.)

Vapour pressure

OECD Test Guideline 104: Vapour Pressure (2006)

(A.4)

Surface tension

OECD Test Guideline 115: Surface Tension of Aqueous Solutions (1995)

A.5.

Water solubility

OECD Test Guideline 105: Water Solubility (1995)

A.6.

Partition coefficient n-octanol/water

OECD Test Guideline 107: Partition Coefficient (n-octanol/water): Shake-Flask Method (1995)

(A.8.)

OECD Test Guideline 123: Partition Coefficient (1-Octanol/Water): Slow-Stirring Method (2022)

A.23.

OECD Test Guideline 117: Partition Coefficient (n-octanol/water): HPLC Method (2022)

A.24.

Dissociation constant

OECD Test Guideline 112: Dissociation Constants in Water. (1981)

A.25.

Viscosity

OECD Test Guideline 114: Viscosity of Liquids (2012)

Flash point

Test methods according to table 2.6.3 of Annex I, Part 2 of Regulation (EC) No 1272/2008

Lower and upper explosion limit

EN 1839:2017 – Determination of the explosion limits and the limiting oxygen concentration (LOC) for flammable gases and vapours

Flammability

Test methods according to section 2.2.4.1. of Annex I, Part 2 of Regulation (EC) No 1272/2008

Test L.2: sustained combustibility test, Part III, section 32 of the UN RTDG Manual of Tests and Criteria

Test N.1: test method for readily combustible solids, Part III, sub-section 33.2.4 of the UN RTDG Manual of Tests and Criteria

Test N.5: test method for substances which in contact with water emit flammable gases, Part III, sub-section 33.5.4 of the UN RTDG Manual of Tests and Criteria

Self-ignition temperature (solids)

Test N.4: test method for self-heating substances, Part III, sub-section 33.4.6 of the UN RTDG Manual of Tests and Criteria

EN 15188:2020 – Determination of the spontanous ignition behaviour of dust accumulations

Auto-ignition temperature (liquids, gases)

ISO/IEC 80079-20-1:2017 – Explosive atmospheres – Part 20-1: Material characteristics for gas and vapour classification – Test methods and data

Decomposition temperature

Test Series H, part II, section 28, of the UN RTDG Manual of Tests and Criteria

Explosive properties

Test methods according to Test series 1-3, Part I, sections 11-13 of the UN RTDG Manual of Tests and Criteria

EU Test method A.14 Explosive Properties

A.14

Oxidising properties

Test method according to section 2.4.4. of Annex I, Part 2 of Regulation (EC) No 1272/2008

Test O.2: test for oxidizing liquids, Part III, sub-section 34.4.2 of the UN RTDG Manual of Tests and Criteria

Test O.1: Test for oxidizing solids, Part III, sub-section 34.4.1 of the UN RTDG Manual of Tests and Criteria

Test O.3 Gravimetric test for oxidizing solids, Part III, sub-section 34.4 3 of the UN RTDG Manual of Tests and Criteria

Pyrophoricity

Test N.3: test method for pyrophoric liquids, Part III, sub-section 33.3.1.5 of the UN RTDG Manual of Tests and Criteria

Test N.2: test method for pyrophoric solids, Part III, sub-section 33.3.1.4 of the UN RTDG Manual of Tests and Criteria

Granulometry/particle characteristics

EU test method A.22. Length Weighted Geometric Mean Diameter of Fibres

A.22.

ISO 13318 – Determination of Particle Size Distribution by Centrifugal Liquid Sedimentation Methods

ISO 21501 – Determination of Particle Size Distribution – Single Particle Light Interaction Methods

OECD Test Guideline 124: Determination of the Volume Specific Surface Area of Manufactured Nanomaterials (2022)

OECD Test Guideline 125: Particle Size and Particle Size Distribution of Nanomaterials (2022)

pH

OECD Test Guideline 122: Determination of pH, Acidity and Alkalinity (2013)

Properties of polymers

OECD Test Guideline 118: Determination of the Number-Average Molecular Weight and the Molecular Weight Distribution of Polymers using Gel Permeation Chromatography (1996)

A.18.

OECD Test Guideline 119: Determination of the Low Molecular Weight Content of a Polymer Using Gel Permeation Chromatography (1996)

A.19.

OECD Test Guideline 120: Solution/Extraction Behaviour of Polymers in Water (2000)

(A.20.)

TABLE 2: TEST METHODS FOR TOXICOLOGICAL PROPERTIES

| Endpoint | Test method | Corresponding chapter, containing the full description of the test method, in Part B of this Annex (numbers in brackets indicate that a chapter, containing the full description of the test method, has been deleted from Part B; empty cell: no corresponding EU test method in Part B of this Annex) |
| --- | --- | --- |
| Skin corrosion/irritation | In vitro: |
| OECD Test Guideline 430: In vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method (TER) (2015) | B.40. |
| OECD Test Guideline 431: In vitro Skin Corrosion: Reconstructed Human Epidermis (RhE) Test Method (2019) | (B.40bis.) |
| OECD Test Guideline 435: In vitro Membrane Barrier Test Method for Skin Corrosion (2015) | B.65. |
| OECD Test Guideline 439: In vitro Skin Irritation: Reconstructed Human Epidermis Test Method (2021) | (B.46.) |
| In vivo: |
| OECD Test Guideline 404: Acute Dermal Irritation/Corrosion (2015) | B.4. |
| Serious eye damage/eye irritation | In vitro: |
| OECD Test Guideline 437: Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2020) | (B.47.) |
| OECD Test Guideline 438: Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2018) | (B.48.) |
| OECD Test Guideline 460: Fluorescein Leakage Test Method for Identifying Ocular Corrosives and Severe Irritants (2017) | (B.61.) |
| OECD Test Guideline 491: Short Time Exposure In Vitro Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2020) | (B.68.) |
| OECD Test Guideline 492: Reconstructed Human Cornea-Like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage (2019) | (B.69.) |
| OECD Test Guideline 492B: Reconstructed Human Cornea-like Epithelium (RHCE) Test Method for Eye Hazard Identification (2022) | |
| OECD Test Guideline 494: Vitrigel-Eye Irritancy Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage (2021) | |
| OECD Test Guideline 496: In vitro Macromolecular Test Method for Identifying Chemicals Inducing Serious Eye Damage and Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2019) | |
| OECD Test Guideline 467: Defined Approaches for Serious Eye Damage and Eye Irritation (2022) | |
| In vivo: |
| OECD Test Guideline 405: Acute Eye Irritation/Corrosion (2021) | (B.5.) |
| Skin sensitisation | In vitro: |
| OECD Test Guideline 442C: In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA) (2022) | (B.59.) |
| OECD Test Guideline 442D: In Vitro Skin Sensitisation Assays Addressing the AOP Key Event on Keratinocyte Activation (2022) | (B.60.) |
| OECD Test Guideline 442E: In Vitro Skin Sensitisation: In Vitro Skin Sensitisation Assays Addressing the Key Event on Activation of Dendritic Cells on the Adverse Outcome Pathway for Skin Sensitisation (2022) | (B.71.) |
| OECD Test Guideline 497: Defined Approaches on Skin Sensitisation (2021) | |
| In vivo: |
| OECD Test Guideline 429: Skin Sensitisation – Local Lymph Node Assay (2010) | B.42. |
| OECD Test Guideline 442A: Skin Sensitisation – Local Lymph Node Assay: DA (2010) | B.50. |
| OECD Test Guideline 442B: Skin Sensitisation – Local Lymph Node Assay: BrdU-ELISA or –FCM (2018) | (B.51.) |
| OECD Test Guideline 406: Skin Sensitisation Guinea Pig
  Maximisation Test and Buehler Test (2022) | (B.6.) |
| Mutagenicity | In vitro: |
| OECD Test Guideline 471: Bacterial Reverse Mutation Test (2020) | (B.13./14.) |
| OECD Test Guideline 476: In Vitro Mammalian Cell Gene Mutation Test Using the Hprt and xprt Genes (2016) | (B.17.) |
| OECD Test Guideline 490: In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene (2016) | B.67. |
| OECD Test Guideline 473: In vitro Mammalian Chromosome Aberration Test (2016) | B.10. |
| OECD Test Guideline 487. In vitro Mammalian Cell Micronucleus Test (2016) | B.49. |
| In vivo: |
| OECD Test Guideline 475: Mammalian Bone Marrow Chromosome Aberration Test (2016) | B.11. |
| OECD Test Guideline 474: Mammalian Erythrocyte Micronucleus Test (2016) | B.12. |
| OECD Test Guideline 483: Mammalian Spermatogonial Chromosome Aberration Test (2016) | B.23. |
| OECD Test Guideline 488: Transgenic Rodent Somatic and Germ Cell Gene Mutation Assays (2022) | (B.58.) |
| OECD Test Guideline 489: In Vivo Mammalian Alkaline Comet Assay (2016) | B.62. |
| OECD Test Guideline 470: Mammalian Erythrocyte Pig-a Gene mutation Assay (2022) | |
| Acute toxicity | Oral: |
| OECD Test Guideline 420: Acute Oral Toxicity: Fixed Dose Procedure (2002) | B.1 bis. |
| OECD Test Guideline 423: Acute Oral Toxicity: Acute Toxic Class Method (2002) | B.1 tris. |
| OECD Test Guideline 425: Acute Oral Toxicity: Up-and-Down Procedure (2022) | |
| Dermal: |
| OECD Test Guideline 402: Acute Dermal Toxicity – Fixed Dose Procedure (2017) | (B.3.) |
| Inhalation: |
| OECD Test Guideline 403: Acute Inhalation Toxicity (2009) | B.2. |
| OECD Test Guideline 436: Acute Inhalation Toxicity – Acute Toxic Class Method (2009) | B.52. |
| OECD Test Guideline 433: Acute Inhalation Toxicity: Fixed Concentration Procedure (2018) | |
| Repeated dose toxicity | OECD Test Guideline 407: Repeated Dose 28-Day Oral Toxicity Study in Rodents (2008) | B.7. |
| OECD Test Guideline 412: Subacute Inhalation Toxicity: 28-Day Study (2018) | (B.8.) |
| OECD Test Guideline 410: Repeated Dose Dermal Toxicity: 21/28-Day Study (1981) | B.9. |
| OECD Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (2016) | B.64. |
| OECD Test Guideline 408: Repeated Dose 90-Day Oral Toxicity Study in Rodents (2018) | (B.26.) |
| OECD Test Guideline 409: Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents (1998) | B.27. |
| OECD Test Guideline 413: Subchronic Inhalation Toxicity: 90-Day Study (2018) | (B.29.) |
| OECD Test Guideline 411: Subchronic Dermal Toxicity: 90-Day Study (1981) | B.28. |
| OECD Test Guideline 452: Chronic Toxicity Studies (2018) | (B.30.) |
| OECD Test Guideline 453: Combined Chronic Toxicity/Carcinogenicity Studies (2018) | (B.33.) |
| Reproductive/developmental toxicity | OECD Test Guideline 443: Extended One-Generation Reproduction Toxicity Study (2018) | (B.56.) |
| OECD Test Guideline 421: Reproduction/Developmental Toxicity Screening Test (2016) | B.63. |
| OECD Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (2016) | B.64. |
| OECD Test Guideline 414: Prenatal Developmental Toxicity Study (2018) | (B.31.) |
| Toxicokinetics | OECD Test Guideline 417: Toxicokinetics (2010) | B.36. |
| OECD Test Guideline 428: Skin Absorption: In Vitro Method (2004) | B.45. |
| OECD Test Guideline 427: Skin Absorption: In Vivo Method (2004) | B.44. |
| Carcinogenicity | OECD Test Guideline 451: Carcinogenicity Studies (2018) | (B.32.) |
| OECD Test Guideline 453: Combined Chronic Toxicity/Carcinogenicity Studies (2018) | (B.33.) |
| EU test method B.21. In Vitro Mammalian Cell Transformation Test | B.21. |
| (Developmental) Neurotoxicity | OECD Test Guideline 424: Neurotoxicity Study in Rodents (1997) | B.43. |
| OECD Test Guideline 426: Developmental Neurotoxicity Study (2007) | B.53. |
| OECD Test Guideline 418: Delayed Neurotoxicity of Organophosphorus Substances Following Acute Exposure (1995) | B.37. |
| OECD Test Guideline 419: Delayed Neurotoxicity of Organophosphorus Substances: 28-day Repeated Dose Study (1995) | B.38. |
| Endocrine disrupting properties | In vitro |
| OECD Test Guideline 455: Performance-Based Test Guideline for Stably Transfected Transactivation In Vitro Assays to Detect Estrogen Receptor Agonists and Antagonistsals (2021) | (B.66.) |
| OECD Test Guideline 456: H295R Steroidogenesis Assay (2022) | B.57. |
| OECD Test Guideline 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals (2020) | |
| OECD Test Guideline 493: Performance-Based Test Guideline for Human Recombinant Estrogen Receptor (hrER) In Vitro Assays to Detect Chemicals with ER Binding Affinity (2015) | B.70. |
| In vivo |
| OECD Test Guideline 440: Uterotrophic Bioassay in Rodents A short-term screening test for oestrogenic properties (2007) | B.54. |
| OECD Test Guideline 441: Hershberger Bioassay in Rats, A Short-term Screening Assay for (Anti)Androgenic Properties (2009) | B.55. |
| Phototoxicity | OECD Test Guideline 432: In Vitro 3T3 NRU Phototoxicity Test (2019) | (B.41.) |
| OECD Test Guideline 495: Ros (Reactive Oxygen Species) Assay for Photoreactivity (2019) | |
| OECD Test Guideline 498: In Vitro Phototoxicity Test Method Using the Reconstructed Human Epidermis (RhE) (2021) | |

Endpoint

Test method

Corresponding chapter, containing the full description of the test method, in Part B of this Annex (numbers in brackets indicate that a chapter, containing the full description of the test method, has been deleted from Part B; empty cell: no corresponding EU test method in Part B of this Annex)

Skin corrosion/irritation

In vitro:

OECD Test Guideline 430: In vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method (TER) (2015)

B.40.

OECD Test Guideline 431: In vitro Skin Corrosion: Reconstructed Human Epidermis (RhE) Test Method (2019)

(B.40bis.)

OECD Test Guideline 435: In vitro Membrane Barrier Test Method for Skin Corrosion (2015)

B.65.

OECD Test Guideline 439: In vitro Skin Irritation: Reconstructed Human Epidermis Test Method (2021)

(B.46.)

In vivo:

OECD Test Guideline 404: Acute Dermal Irritation/Corrosion (2015)

B.4.

Serious eye damage/eye irritation

In vitro:

OECD Test Guideline 437: Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2020)

(B.47.)

OECD Test Guideline 438: Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2018)

(B.48.)

OECD Test Guideline 460: Fluorescein Leakage Test Method for Identifying Ocular Corrosives and Severe Irritants (2017)

(B.61.)

OECD Test Guideline 491: Short Time Exposure In Vitro Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2020)

(B.68.)

OECD Test Guideline 492: Reconstructed Human Cornea-Like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage (2019)

(B.69.)

OECD Test Guideline 492B: Reconstructed Human Cornea-like Epithelium (RHCE) Test Method for Eye Hazard Identification (2022)

OECD Test Guideline 494: Vitrigel-Eye Irritancy Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage (2021)

OECD Test Guideline 496: In vitro Macromolecular Test Method for Identifying Chemicals Inducing Serious Eye Damage and Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2019)

OECD Test Guideline 467: Defined Approaches for Serious Eye Damage and Eye Irritation (2022)

In vivo:

OECD Test Guideline 405: Acute Eye Irritation/Corrosion (2021)

(B.5.)

Skin sensitisation

In vitro:

OECD Test Guideline 442C: In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA) (2022)

(B.59.)

OECD Test Guideline 442D: In Vitro Skin Sensitisation Assays Addressing the AOP Key Event on Keratinocyte Activation (2022)

(B.60.)

OECD Test Guideline 442E: In Vitro Skin Sensitisation: In Vitro Skin Sensitisation Assays Addressing the Key Event on Activation of Dendritic Cells on the Adverse Outcome Pathway for Skin Sensitisation (2022)

(B.71.)

OECD Test Guideline 497: Defined Approaches on Skin Sensitisation (2021)

In vivo:

OECD Test Guideline 429: Skin Sensitisation – Local Lymph Node Assay (2010)

B.42.

OECD Test Guideline 442A: Skin Sensitisation – Local Lymph Node Assay: DA (2010)

B.50.

OECD Test Guideline 442B: Skin Sensitisation – Local Lymph Node Assay: BrdU-ELISA or –FCM (2018)

(B.51.)

OECD Test Guideline 406: Skin Sensitisation Guinea Pig

Maximisation Test and Buehler Test (2022)

(B.6.)

Mutagenicity

In vitro:

OECD Test Guideline 471: Bacterial Reverse Mutation Test (2020)

(B.13./14.)

OECD Test Guideline 476: In Vitro Mammalian Cell Gene Mutation Test Using the Hprt and xprt Genes (2016)

(B.17.)

OECD Test Guideline 490: In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene (2016)

B.67.

OECD Test Guideline 473: In vitro Mammalian Chromosome Aberration Test (2016)

B.10.

OECD Test Guideline 487. In vitro Mammalian Cell Micronucleus Test (2016)

B.49.

In vivo:

OECD Test Guideline 475: Mammalian Bone Marrow Chromosome Aberration Test (2016)

B.11.

OECD Test Guideline 474: Mammalian Erythrocyte Micronucleus Test (2016)

B.12.

OECD Test Guideline 483: Mammalian Spermatogonial Chromosome Aberration Test (2016)

B.23.

OECD Test Guideline 488: Transgenic Rodent Somatic and Germ Cell Gene Mutation Assays (2022)

(B.58.)

OECD Test Guideline 489: In Vivo Mammalian Alkaline Comet Assay (2016)

B.62.

OECD Test Guideline 470: Mammalian Erythrocyte Pig-a Gene mutation Assay (2022)

Acute toxicity

Oral:

OECD Test Guideline 420: Acute Oral Toxicity: Fixed Dose Procedure (2002)

B.1 bis.

OECD Test Guideline 423: Acute Oral Toxicity: Acute Toxic Class Method (2002)

B.1 tris.

OECD Test Guideline 425: Acute Oral Toxicity: Up-and-Down Procedure (2022)

Dermal:

OECD Test Guideline 402: Acute Dermal Toxicity – Fixed Dose Procedure (2017)

(B.3.)

Inhalation:

OECD Test Guideline 403: Acute Inhalation Toxicity (2009)

B.2.

OECD Test Guideline 436: Acute Inhalation Toxicity – Acute Toxic Class Method (2009)

B.52.

OECD Test Guideline 433: Acute Inhalation Toxicity: Fixed Concentration Procedure (2018)

Repeated dose toxicity

OECD Test Guideline 407: Repeated Dose 28-Day Oral Toxicity Study in Rodents (2008)

B.7.

OECD Test Guideline 412: Subacute Inhalation Toxicity: 28-Day Study (2018)

(B.8.)

OECD Test Guideline 410: Repeated Dose Dermal Toxicity: 21/28-Day Study (1981)

B.9.

OECD Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (2016)

B.64.

OECD Test Guideline 408: Repeated Dose 90-Day Oral Toxicity Study in Rodents (2018)

(B.26.)

OECD Test Guideline 409: Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents (1998)

B.27.

OECD Test Guideline 413: Subchronic Inhalation Toxicity: 90-Day Study (2018)

(B.29.)

OECD Test Guideline 411: Subchronic Dermal Toxicity: 90-Day Study (1981)

B.28.

OECD Test Guideline 452: Chronic Toxicity Studies (2018)

(B.30.)

OECD Test Guideline 453: Combined Chronic Toxicity/Carcinogenicity Studies (2018)

(B.33.)

Reproductive/developmental toxicity

OECD Test Guideline 443: Extended One-Generation Reproduction Toxicity Study (2018)

(B.56.)

OECD Test Guideline 421: Reproduction/Developmental Toxicity Screening Test (2016)

B.63.

OECD Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (2016)

B.64.

OECD Test Guideline 414: Prenatal Developmental Toxicity Study (2018)

(B.31.)

Toxicokinetics

OECD Test Guideline 417: Toxicokinetics (2010)

B.36.

OECD Test Guideline 428: Skin Absorption: In Vitro Method (2004)

B.45.

OECD Test Guideline 427: Skin Absorption: In Vivo Method (2004)

B.44.

Carcinogenicity

OECD Test Guideline 451: Carcinogenicity Studies (2018)

(B.32.)

OECD Test Guideline 453: Combined Chronic Toxicity/Carcinogenicity Studies (2018)

(B.33.)

EU test method B.21. In Vitro Mammalian Cell Transformation Test

B.21.

(Developmental) Neurotoxicity

OECD Test Guideline 424: Neurotoxicity Study in Rodents (1997)

B.43.

OECD Test Guideline 426: Developmental Neurotoxicity Study (2007)

B.53.

OECD Test Guideline 418: Delayed Neurotoxicity of Organophosphorus Substances Following Acute Exposure (1995)

B.37.

OECD Test Guideline 419: Delayed Neurotoxicity of Organophosphorus Substances: 28-day Repeated Dose Study (1995)

B.38.

Endocrine disrupting properties

In vitro

OECD Test Guideline 455: Performance-Based Test Guideline for Stably Transfected Transactivation In Vitro Assays to Detect Estrogen Receptor Agonists and Antagonistsals (2021)

(B.66.)

OECD Test Guideline 456: H295R Steroidogenesis Assay (2022)

B.57.

OECD Test Guideline 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals (2020)

OECD Test Guideline 493: Performance-Based Test Guideline for Human Recombinant Estrogen Receptor (hrER) In Vitro Assays to Detect Chemicals with ER Binding Affinity (2015)

B.70.

In vivo

OECD Test Guideline 440: Uterotrophic Bioassay in Rodents A short-term screening test for oestrogenic properties (2007)

B.54.

OECD Test Guideline 441: Hershberger Bioassay in Rats, A Short-term Screening Assay for (Anti)Androgenic Properties (2009)

B.55.

Phototoxicity

OECD Test Guideline 432: In Vitro 3T3 NRU Phototoxicity Test (2019)

(B.41.)

OECD Test Guideline 495: Ros (Reactive Oxygen Species) Assay for Photoreactivity (2019)

OECD Test Guideline 498: In Vitro Phototoxicity Test Method Using the Reconstructed Human Epidermis (RhE) (2021)

TABLE 3: TEST METHODS FOR ECOTOXICOLOGICAL PROPERTIES

| Endpoint | Test method | Corresponding chapter in Part C, containing the full description of the test method, of this Annex (numbers in brackets indicate that a chapter, containing the full description of the test method, has been deleted from Part C; empty cell: no corresponding EU test method in Part C of this Annex) |
| --- | --- | --- |
| Aquatic toxicity | OECD Test Guideline 201: Freshwater Alga and Cyanobacteria, Growth Inhibition Test (2011) | C.3. |
| OECD Test Guideline 209: Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation) (2010) | C.11. |
| OECD Test Guideline 224: Determination of the Inhibition of the Activity of Anaerobic Bacteria (2007) | C.34. |
| OECD Test Guideline 244: Protozoan Activated Sludge Inhibition Test (2017) | |
| OECD Test Guideline 221: Lemna sp. Growth Inhibition Test (2006) | C.26. |
| OECD Test Guideline 202: Daphnia sp. Acute Immobilisation Test (2004) | C.2. |
| OECD Test Guideline 211: Daphnia magna Reproduction Test (2012) | C.20. |
| OECD Test Guideline 203: Fish, Acute Toxicity Test (2019) | (C.1.) |
| OECD Test Guideline 210: Fish, Early-life Stage Toxicity Test (2013) | C.47. |
| OECD Test Guideline 215: Fish, Juvenile Growth Test (2000) | C.14. |
| OECD Test Guideline 236: Fish Embryo Acute Toxicity (FET) Test (2013) | C.49. |
| OECD Test Guideline 249: Fish Cell Line Acute Toxicity – the RTgill-W1 Cell Line Assay (2021) | |
| OECD Test Guideline 242: Potamopyrgus antipodarum Reproduction Test (2016) | |
| OECD Test Guideline 243: Lymnaea stagnalis Reproduction Test (2016) | |
| Degradation | OECD Test Guideline 111: Hydrolysis as a Function of pH (2004) | C.7. |
| OECD Test Guideline 301: Ready Biodegradability (1992) | C.4. |
| OECD Test Guideline 302A: Inherent Biodegradability: Modified SCAS Test (1981) | C.12. |
| OECD Test Guideline 302B: Inherent Biodegradability: Zahn-Wellens/EMPA Test (1992) | (C.9). |
| OECD Test Guideline 302C: Inherent Biodegradability: Modified MITI Test (II) (2009) | |
| OECD Test Guideline 303: Simulation Test – Aerobic Sewage Treatment — A: Activated Sludge Units; B: Biofilms (2001) | C.10. |
| OECD Test Guideline 304A: Inherent Biodegradability in Soil (1981) | |
| OECD Test Guideline 306: Biodegradability in Seawater (1992) | C.42. |
| OECD Test Guideline 307: Aerobic and Anaerobic Transformation in Soil (2002) | C.23. |
| OECD Test Guideline 308: Aerobic and Anaerobic Transformation in Aquatic Sediment Systems (2002) | C.24. |
| OECD Test Guideline 309: Aerobic Mineralisation in Surface Water – Simulation Biodegradation Test (2004) | C.25. |
| OECD Test Guideline 310: Ready Biodegradability – CO2 in sealed vessels (Headspace Test) (2014) | C.29. |
| OECD Test Guideline 311: Anaerobic Biodegradability of Organic Compounds in Digested Sludge: by Measurement of Gas Production (2006) | C.43. |
| OECD Test Guideline 314: Simulation Tests to Assess the Biodegradability of Chemicals Discharged in Wastewater (2008) | |
| OECD Test Guideline 316: Phototransformation of Chemicals in Water – Direct Photolysis (2008) | |
| EU test method C.5. Degradation – Biochemical Oxygen Demand | C.5. |
| EU test method C.6. Degradation – Chemical Oxygen Demand | C.6. |
| Fate and behaviour in the environment | OECD Test Guideline 305: Bioaccumulation in Fish: Aqueous and Dietary Exposure (2012) | C.13. |
| OECD Test Guideline 315: Bioaccumulation in Sediment-Dwelling Benthic Oligochaetes (2008) | C.46. |
| OECD Test Guideline 317: Bioaccumulation in Terrestrial Oligochaetes (2010) | C.30. |
| OECD Test Guideline 318: Dispersion Stability of Nanomaterials in Simulated Environmental Media (2017) | |
| OECD Test Guideline 121: Estimation of the Adsorption Coefficient (Koc) on Soil and on Sewage Sludge using High Performance Liquid Chromatography (HPLC) (2001) | C.19. |
| OECD Test Guideline 106: Adsorption – Desorption Using a Batch Equilibrium Method (2000) | C.18. |
| OECD Test Guideline 312: Leaching in Soil Columns (2004) | C.44. |
| OECD Test Guideline 313: Estimation of Emissions from Preservative – Treated Wood to the Environment (2007) | C.45. |
| OECD Test Guideline 319A: Determination of In Vitro Intrinsic Clearance Using Cryopreserved Rainbow Trout Hepatocytes (RT-HEP) (2018) | |
| OECD Test Guideline 319B: Determination of In Vitro Intrinsic Clearance Using Rainbow Trout Liver S9 Sub-Cellular Fraction (RT-S9) (2018) | |
| OECD Test Guideline 320: Anaerobic Transformation of Chemicals in Liquid Manure (2022) | |
| Effects on terrestrial organisms | OECD Test Guideline 216: Soil Microorganisms: Nitrogen Transformation Test (2000) | C.21. |
| OECD Test Guideline 217: Soil Microorganisms: Carbon Transformation Test (2000) | C.22. |
| OECD Test Guideline 207: Earthworm, Acute Toxicity Tests (1984) | C.8. |
| OECD Test Guideline 222: Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei) (2016) | (C.33.) |
| OECD Test Guideline 220: Enchytraeid Reproduction Test (2016) | (C.32.) |
| OECD Test Guideline 226: Predatory Mite (Hypoaspis (Geolaelaps) aculeifer) Reproduction Test in Soil (2016) | (C.36.) |
| OECD Test Guideline 232: Collembolan Reproduction Test in Soil (2016) | (C.39.) |
| OECD Test Guideline 208: Terrestrial Plant Test: Seedling Emergence and Seedling Growth Test (2006) | C.31. |
| OECD Test Guideline 227: Terrestrial Plant Test: Vegetative Vigour Test (2006) | |
| Effects on sediment organisms | OECD Test Guideline 218: Sediment-Water Chironomid Toxicity Using Spiked Sediment (2004) | C.27. |
| OECD Test Guideline 219: Sediment-Water Chironomid Toxicity Using Spiked Water (2004) | C.28. |
| OECD Test Guideline 233: Sediment-Water Chironomid Life-Cycle Toxicity Test Using Spiked Water or Spiked Sediment (2010) | C.40. |
| OECD Test Guideline 235: Chironomus sp., Acute Immobilisation Test (2011) | |
| OECD Test Guideline 225: Sediment-Water Lumbriculus Toxicity Test Using Spiked Sediment (2007) | C.35. |
| OECD Test Guideline 238: Sediment-Free Myriophyllum Spicatum Toxicity Test (2014) | C.50. |
| OECD Test Guideline 239: Water-Sediment Myriophyllum Spicatum Toxicity Test (2014) | C.51. |
| Effects on birds | OECD Test Guideline 205: Avian Dietary Toxicity Test (1984) | |
| OECD Test Guideline 206: Avian Reproduction Test (1984) | |
| OECD Test Guideline 223: Avian Acute Oral Toxicity Test (2016) | |
| Effects on insects | OECD Test Guideline 213: Honeybees, Acute Oral Toxicity Test (1998) | C.16. |
| OECD Test Guideline 214: Honeybees, Acute Contact Toxicity Test (1998) | C.17. |
| OECD Test Guideline 237: Honey Bee (Apis Mellifera) Larval Toxicity Test, Single Exposure (2013) | |
| OECD Test Guideline 245: Honey Bee (Apis Mellifera L.), Chronic Oral Toxicity Test (10-Day Feeding) (2017) | |
| OECD Test Guideline 246: Bumblebee, Acute Contact Toxicity Test (2017) | |
| OECD Test Guideline 247: Bumblebee, Acute Oral Toxicity Test (2017) | |
| OECD Test Guideline 228: Determination of Developmental Toxicity to Dipteran Dung Flies (Scathophaga stercoraria L. (Scathophagidae), Musca autumnalis De Geer (Muscidae)) (2016) | |
| Endocrine disrupting properties | OECD Test Guideline 230: 21-Day Fish Assay (2009) | C.37. |
| OECD Test Guideline 229: Fish Short Term Reproduction Assay (2012) | C.48. |
| OECD Test Guideline 231: Amphibian Metamorphosis Assay (2009) | C.38. |
| OECD Test Guideline 234: Fish Sexual Development Test (2011) | C.41. |
| OECD Test Guideline 240: Medaka Extended OneGeneration Reproduction Test (MEOGRT) (2015) | C.52. |
| OECD Test Guideline 241: The Larval Amphibian Growth and Development Assay (LAGDA) (2015) | C.53.’ |
| OECD Test Guideline 248: Xenopus Eleutheroembryonic Thyroid Assay (XETA) (2019) | |
| OECD Test Guideline 250: EASZY assay – Detection of Endocrine Active Substances, Acting Through Estrogen Receptors, Using Transgenic tg(cyp19a1b:GFP) Zebrafish embrYos (2021) | |
| OECD Test Guideline 251: Rapid Androgen Disruption Activity Reporter (RADAR) Assay (2022) | |

Endpoint

Test method

Corresponding chapter in Part C, containing the full description of the test method, of this Annex (numbers in brackets indicate that a chapter, containing the full description of the test method, has been deleted from Part C; empty cell: no corresponding EU test method in Part C of this Annex)

Aquatic toxicity

OECD Test Guideline 201: Freshwater Alga and Cyanobacteria, Growth Inhibition Test (2011)

C.3.

OECD Test Guideline 209: Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation) (2010)

C.11.

OECD Test Guideline 224: Determination of the Inhibition of the Activity of Anaerobic Bacteria (2007)

C.34.

OECD Test Guideline 244: Protozoan Activated Sludge Inhibition Test (2017)

OECD Test Guideline 221: Lemna sp. Growth Inhibition Test (2006)

C.26.

OECD Test Guideline 202: Daphnia sp. Acute Immobilisation Test (2004)

C.2.

OECD Test Guideline 211: Daphnia magna Reproduction Test (2012)

C.20.

OECD Test Guideline 203: Fish, Acute Toxicity Test (2019)

(C.1.)

OECD Test Guideline 210: Fish, Early-life Stage Toxicity Test (2013)

C.47.

OECD Test Guideline 215: Fish, Juvenile Growth Test (2000)

C.14.

OECD Test Guideline 236: Fish Embryo Acute Toxicity (FET) Test (2013)

C.49.

OECD Test Guideline 249: Fish Cell Line Acute Toxicity – the RTgill-W1 Cell Line Assay (2021)

OECD Test Guideline 242: Potamopyrgus antipodarum Reproduction Test (2016)

OECD Test Guideline 243: Lymnaea stagnalis Reproduction Test (2016)

Degradation

OECD Test Guideline 111: Hydrolysis as a Function of pH (2004)

C.7.

OECD Test Guideline 301: Ready Biodegradability (1992)

C.4.

OECD Test Guideline 302A: Inherent Biodegradability: Modified SCAS Test (1981)

C.12.

OECD Test Guideline 302B: Inherent Biodegradability: Zahn-Wellens/EMPA Test (1992)

(C.9).

OECD Test Guideline 302C: Inherent Biodegradability: Modified MITI Test (II) (2009)

OECD Test Guideline 303: Simulation Test – Aerobic Sewage Treatment — A: Activated Sludge Units; B: Biofilms (2001)

C.10.

OECD Test Guideline 304A: Inherent Biodegradability in Soil (1981)

OECD Test Guideline 306: Biodegradability in Seawater (1992)

C.42.

OECD Test Guideline 307: Aerobic and Anaerobic Transformation in Soil (2002)

C.23.

OECD Test Guideline 308: Aerobic and Anaerobic Transformation in Aquatic Sediment Systems (2002)

C.24.

OECD Test Guideline 309: Aerobic Mineralisation in Surface Water – Simulation Biodegradation Test (2004)

C.25.

OECD Test Guideline 310: Ready Biodegradability – CO2 in sealed vessels (Headspace Test) (2014)

C.29.

OECD Test Guideline 311: Anaerobic Biodegradability of Organic Compounds in Digested Sludge: by Measurement of Gas Production (2006)

C.43.

OECD Test Guideline 314: Simulation Tests to Assess the Biodegradability of Chemicals Discharged in Wastewater (2008)

OECD Test Guideline 316: Phototransformation of Chemicals in Water – Direct Photolysis (2008)

EU test method C.5. Degradation – Biochemical Oxygen Demand

C.5.

EU test method C.6. Degradation – Chemical Oxygen Demand

C.6.

Fate and behaviour in the environment

OECD Test Guideline 305: Bioaccumulation in Fish: Aqueous and Dietary Exposure (2012)

C.13.

OECD Test Guideline 315: Bioaccumulation in Sediment-Dwelling Benthic Oligochaetes (2008)

C.46.

OECD Test Guideline 317: Bioaccumulation in Terrestrial Oligochaetes (2010)

C.30.

OECD Test Guideline 318: Dispersion Stability of Nanomaterials in Simulated Environmental Media (2017)

OECD Test Guideline 121: Estimation of the Adsorption Coefficient (Koc) on Soil and on Sewage Sludge using High Performance Liquid Chromatography (HPLC) (2001)

C.19.

OECD Test Guideline 106: Adsorption – Desorption Using a Batch Equilibrium Method (2000)

C.18.

OECD Test Guideline 312: Leaching in Soil Columns (2004)

C.44.

OECD Test Guideline 313: Estimation of Emissions from Preservative – Treated Wood to the Environment (2007)

C.45.

OECD Test Guideline 319A: Determination of In Vitro Intrinsic Clearance Using Cryopreserved Rainbow Trout Hepatocytes (RT-HEP) (2018)

OECD Test Guideline 319B: Determination of In Vitro Intrinsic Clearance Using Rainbow Trout Liver S9 Sub-Cellular Fraction (RT-S9) (2018)

OECD Test Guideline 320: Anaerobic Transformation of Chemicals in Liquid Manure (2022)

Effects on terrestrial organisms

OECD Test Guideline 216: Soil Microorganisms: Nitrogen Transformation Test (2000)

C.21.

OECD Test Guideline 217: Soil Microorganisms: Carbon Transformation Test (2000)

C.22.

OECD Test Guideline 207: Earthworm, Acute Toxicity Tests (1984)

C.8.

OECD Test Guideline 222: Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei) (2016)

(C.33.)

OECD Test Guideline 220: Enchytraeid Reproduction Test (2016)

(C.32.)

OECD Test Guideline 226: Predatory Mite (Hypoaspis (Geolaelaps) aculeifer) Reproduction Test in Soil (2016)

(C.36.)

OECD Test Guideline 232: Collembolan Reproduction Test in Soil (2016)

(C.39.)

OECD Test Guideline 208: Terrestrial Plant Test: Seedling Emergence and Seedling Growth Test (2006)

C.31.

OECD Test Guideline 227: Terrestrial Plant Test: Vegetative Vigour Test (2006)

Effects on sediment organisms

OECD Test Guideline 218: Sediment-Water Chironomid Toxicity Using Spiked Sediment (2004)

C.27.

OECD Test Guideline 219: Sediment-Water Chironomid Toxicity Using Spiked Water (2004)

C.28.

OECD Test Guideline 233: Sediment-Water Chironomid Life-Cycle Toxicity Test Using Spiked Water or Spiked Sediment (2010)

C.40.

OECD Test Guideline 235: Chironomus sp., Acute Immobilisation Test (2011)

OECD Test Guideline 225: Sediment-Water Lumbriculus Toxicity Test Using Spiked Sediment (2007)

C.35.

OECD Test Guideline 238: Sediment-Free Myriophyllum Spicatum Toxicity Test (2014)

C.50.

OECD Test Guideline 239: Water-Sediment Myriophyllum Spicatum Toxicity Test (2014)

C.51.

Effects on birds

OECD Test Guideline 205: Avian Dietary Toxicity Test (1984)

OECD Test Guideline 206: Avian Reproduction Test (1984)

OECD Test Guideline 223: Avian Acute Oral Toxicity Test (2016)

Effects on insects

OECD Test Guideline 213: Honeybees, Acute Oral Toxicity Test (1998)

C.16.

OECD Test Guideline 214: Honeybees, Acute Contact Toxicity Test (1998)

C.17.

OECD Test Guideline 237: Honey Bee (Apis Mellifera) Larval Toxicity Test, Single Exposure (2013)

OECD Test Guideline 245: Honey Bee (Apis Mellifera L.), Chronic Oral Toxicity Test (10-Day Feeding) (2017)

OECD Test Guideline 246: Bumblebee, Acute Contact Toxicity Test (2017)

OECD Test Guideline 247: Bumblebee, Acute Oral Toxicity Test (2017)

OECD Test Guideline 228: Determination of Developmental Toxicity to Dipteran Dung Flies (Scathophaga stercoraria L. (Scathophagidae), Musca autumnalis De Geer (Muscidae)) (2016)

Endocrine disrupting properties

OECD Test Guideline 230: 21-Day Fish Assay (2009)

C.37.

OECD Test Guideline 229: Fish Short Term Reproduction Assay (2012)

C.48.

OECD Test Guideline 231: Amphibian Metamorphosis Assay (2009)

C.38.

OECD Test Guideline 234: Fish Sexual Development Test (2011)

C.41.

OECD Test Guideline 240: Medaka Extended OneGeneration Reproduction Test (MEOGRT) (2015)

C.52.

OECD Test Guideline 241: The Larval Amphibian Growth and Development Assay (LAGDA) (2015)

C.53.’

OECD Test Guideline 248: Xenopus Eleutheroembryonic Thyroid Assay (XETA) (2019)

OECD Test Guideline 250: EASZY assay – Detection of Endocrine Active Substances, Acting Through Estrogen Receptors, Using Transgenic tg(cyp19a1b:GFP) Zebrafish embrYos (2021)

OECD Test Guideline 251: Rapid Androgen Disruption Activity Reporter (RADAR) Assay (2022)

| (2) | in part A, the text below the title of each of the Chapters A.3., A.4., A.8. to A.12., A.15. to A.17., A.20. and A.21. is replaced by: ‘The full description of this test method has been deleted. The equivalent international test method, or other applicable test methods for the endpoint in question, appear in Table 1 of Part 0.’ |
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(2)

in part A, the text below the title of each of the Chapters A.3., A.4., A.8. to A.12., A.15. to A.17., A.20. and A.21. is replaced by: ‘The full description of this test method has been deleted. The equivalent international test method, or other applicable test methods for the endpoint in question, appear in Table 1 of Part 0.’

| (3) | in part B, the text below the title of each of the Chapters B.3., B.5., B.6., B.8., B. 13/14., B.17., B.26., B.29. to B.33., B.40bis., B.41., B.46. to B.48., B.51., B.56., B.58. to B.61., B.66., B.68., B.69. and B.71. is replaced by: ‘The full description of this test method has been deleted. The equivalent international test method appears in Table 2 of Part 0.’ |
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(3)

in part B, the text below the title of each of the Chapters B.3., B.5., B.6., B.8., B. 13/14., B.17., B.26., B.29. to B.33., B.40bis., B.41., B.46. to B.48., B.51., B.56., B.58. to B.61., B.66., B.68., B.69. and B.71. is replaced by: ‘The full description of this test method has been deleted. The equivalent international test method appears in Table 2 of Part 0.’

| (4) | in part B, the text below the title of each of the Chapters B.22., B.25., B.34., B.35., B.39. is replaced by: ‘This test method has been deleted, as it is no longer recognised as being appropriate for generating information on the toxicological properties of chemicals for the purpose of Regulation (EC) No 1907/2006. The applicable test methods for the endpoint in question appear in Table 2 of Part 0.’ |
| --- | --- |

(4)

in part B, the text below the title of each of the Chapters B.22., B.25., B.34., B.35., B.39. is replaced by: ‘This test method has been deleted, as it is no longer recognised as being appropriate for generating information on the toxicological properties of chemicals for the purpose of Regulation (EC) No 1907/2006. The applicable test methods for the endpoint in question appear in Table 2 of Part 0.’

| (5) | in part C the text below the title of each of the Chapters C.1., C.9., C.32., C.33., C.36. and C.39. is replaced by: ‘The full description of this test method has been deleted. The equivalent international test method appears in Table 3 of Part 0.’ |
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(5)

in part C the text below the title of each of the Chapters C.1., C.9., C.32., C.33., C.36. and C.39. is replaced by: ‘The full description of this test method has been deleted. The equivalent international test method appears in Table 3 of Part 0.’

| (6) | in part C, the text below the title of Chapters C.15 is replaced by: ‘This test method has been deleted, as it is no longer recognised as being appropriate for generating information on the ecotoxicological properties of chemicals for the purpose of Regulation (EC) No 1907/2006. The applicable test methods for the endpoint in question appear in Table 3 of Part 0.’ |
| --- | --- |

(6)

in part C, the text below the title of Chapters C.15 is replaced by: ‘This test method has been deleted, as it is no longer recognised as being appropriate for generating information on the ecotoxicological properties of chemicals for the purpose of Regulation (EC) No 1907/2006. The applicable test methods for the endpoint in question appear in Table 3 of Part 0.’

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