OECD GUIDELINE FOR THE TESTING OF CHEMICALS In Vivo Eye Irritation/Serious Eye Damage

OECD/OCDE 405

Adopted:

9 October 2017

Corrected:

14 June 2021

(introduction updated)

OECD GUIDELINE FOR THE TESTING OF CHEMICALS

In Vivo Eye Irritation/Serious Eye Damage

INTRODUCTION

1. OECD Guidelines for Testing of Chemicals are periodically reviewed to ensure that they reflect the

best available science. In previous reviews of this Test Guideline (TG), special attention was given to

possible improvements through the evaluation of all existing information on the test chemical in order to

avoid unnecessary testing in laboratory animals and thereby address animal welfare concerns. This

updated version of TG 405 (originally adopted in 1981 and updated in 1987, 2002, 2012, 2020 and 2021)

includes reference to the Guidance Document (GD) 263 on Integrated Approaches to Testing and

Assessment (IATA) for Serious Eye Damage and Eye Irritation (1), proposing a modular approach for eye

irritation and serious eye damage testing. The IATA describes several modules which group information

sources and analysis tools, and (i) provides guidance on how to integrate and use existing testing and nontesting

data for the assessment of the eye irritation and serious eye damage potentials of chemicals and

(ii) proposes an approach when further testing is needed (1) The present in vivo rabbit eye test should be

conducted only as a last resort after all the existing information has been considered, and in vitro testing

has been conducted and evaluated as recommended within the IATA for serious eye damage and eye

irritation described within the OECD GD 263. At the time of drafting of this updated TG 405, there are

instances where using this TG may still be necessary or required by some regulatory authorities.

2. The update from 2012 mainly focused on the use of analgesics and anesthetics without impacting

the basic concept and structure of the TG. The Interagency Coordinating Committee on the Validation of

Alternative Methods (ICCVAM) and an independent international scientific peer review panel reviewed the

usefulness and limitations of routinely using topical anesthetics, systemic analgesics, and humane

endpoints during in vivo ocular irritation safety testing (2). The review concluded that the use of topical

anesthetics and systemic analgesics could avoid most or all pain and distress without affecting the outcome

of the test, and recommended that these substances should always be used. This TG takes this review

into account. Topical anesthetics, systemic analgesics, and humane endpoints should be routinely used

during eye irritation and serious eye damage in vivo testing. Exceptions to their use should be justified.

The refinements described in this proposal will substantially reduce or avoid animal pain and distress in

most testing situations where in vivo ocular safety testing is still necessary.

© OECD, (2021)

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3. Balanced preemptive pain management should include (i) routine pretreatment with a topical

anesthetic (e.g., proparacaine or tetracaine) and a systemic analgesic (e.g. buprenorphine), (ii) routine

post-treatment schedule of systemic analgesia (e.g., buprenorphine and meloxicam), (iii) scheduled

observation, monitoring, and recording of animals for clinical signs of pain and/or distress, and (iv)

scheduled observation, monitoring, and recording of the nature, severity, and progression of all eye

injuries. Further detail is provided in the updated procedures described below. Following test chemical

administration, no additional topical anesthetics or analgesics should be applied in order to avoid

interference with the study. Analgesics with anti-inflammatory activity (e.g., meloxicam) should not be

applied topically, and doses used systemically should not interfere with ocular effects.

4. Definitions are set out in the Annex to the TG.

INITIAL CONSIDERATIONS

5. In the interest of both sound science and animal welfare, in vivo testing should not be considered

until all available data relevant to the potential eye irritation/serious eye damage of the test chemical have

been evaluated in a weight-of-the-evidence (WoE) analysis as presented in the GD263 (1) i.e. over the

three Parts of this GD and their corresponding modules (1). Briefly, under Part 1 existing data is addressed

over eight modules covering human data, in vivo data, in vitro/ex vivo eye or skin irritation/corrosion data,

physico-chemical properties data (e.g. pH) and non-testing methods. Under Part 2, WoE analysis is

performed. If this WoE analysis is still inconclusive, Part 3 should be conducted with additional testing,

starting with in vitro methods (3)(4)(5)(6)(7)(8) or in chemico method (9), and in vivo testing is used as last

resort. The in vivo animal test, if e.g. required by regulators, should be considered after conducting in vitro

testing only when: i) the test chemical is not directly identified as UN GHS Cat. 1, UN GHS Cat. 2 or as UN

GHS No Cat. by currently adopted in vitro test methods and defined approaches.ii) WoE assessment

cannot conclude with high enough confidence if the test chemical is Cat. 1, Cat. 2 (or Cat. 2A or Cat. 2B,

if applicable), or No Cat. depending on country-specific regulatory requirements. iii) the test chemical

cannot be tested with the currently available in vitro test methods or defined approaches due to the

limitations of the test methods or when falling outside of the applicability domain of the test method or

approach.”

PRINCIPLE OF THE IN VIVO TEST

6. Following pretreatment with a systemic analgesic and instillation of appropriate topical anesthesia,

the test chemical is applied in a single dose to one of the eyes of the experimental animal; the untreated

eye serves as the control. The degree of eye irritation/serious eye damage is evaluated by scoring lesions

of conjunctiva, cornea, and iris, at specific intervals. Other effects in the eye and adverse systemic effects

are also described to provide a complete evaluation of the effects. The duration of the study should be

sufficient to evaluate the reversibility or irreversibility of the effects.

7. Animals showing signs of severe distress and/or pain at any stage of the test or lesions consistent

with the humane endpoints described in this TG (see Paragraph 24) should be humanely killed, and the

test chemical assessed accordingly without performing further testing with more animals. Criteria for

making the decision to humanely kill moribund and severely suffering animals are the subject of a separate

GD (10).

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PREPARATIONS FOR THE IN VIVO TEST

Selection of species

8. The albino rabbit is the preferable laboratory animal and healthy young adult animals are used. A

rationale for using other strains or species should be provided.

Preparation of animals

9. Both eyes of each experimental animal provisionally selected for testing should be examined within

24 hours before testing starts. Animals showing eye irritation, ocular defects, or pre-existing corneal injury

should not be used.

Housing and feeding conditions

10. Animals should be individually housed. The temperature of the experimental animal room should

be 20°C (± 3°C) for rabbits. Although the relative humidity should be at least 30% and preferably not

exceed 70%, other than during room cleaning, the aim should be 50-60%. Lighting should be artificial, the

sequence being 12 hours light, 12 hours dark. Excessive light intensity should be avoided. For feeding,

conventional laboratory diets may be used with an unrestricted supply of drinking water.

TEST PROCEDURE

Use of topical anesthetics and systemic analgesics

11. The following procedures are recommended to avoid or minimize pain and distress in ocular safety

testing procedures. Alternate procedures that have been determined to provide as good or better

avoidance or relief of pain and distress may be substituted.

• Sixty minutes prior to test chemical application (TCA), buprenorphine 0.01 mg/kg is administered

by subcutaneous injection (SC) to provide a therapeutic level of systemic analgesia.

Buprenorphine and other similar opioid analgesics administered systemically are not known or

expected to alter ocular responses (2).

• Five minutes prior to TCA, one or two drops of a topical ocular anesthetic (e.g. 0.5% proparacaine

hydrochloride or 0.5% tetracaine hydrochloride) are applied to each eye. In order to avoid possible

interference with the study, a topical anesthetic that does not contain preservatives is

recommended. The eye of each animal that is not treated with a test chemical, but which is treated

with topical anesthetics, serves as a control. If the test chemical is anticipated to cause significant

pain and distress, it should not normally be tested in vivo. However, in case of doubt or where

testing is necessary, consideration should be given to additional applications of the topical

anesthetic at 5-minute intervals prior to TCA. Users should be aware that multiple applications of

topical anesthetics could potentially cause a slight increase in the severity and/or time required for

chemically-induced lesions to clear.

• Eight hours after TCA, buprenorphine 0.01 mg/kg SC and meloxicam 0.5 mg/kg SC are

administered to provide a continued therapeutic level of systemic analgesia. While there are no

data to suggest that meloxicam has anti-inflammatory effects on the eye when administered SC

once daily, meloxicam should not be administered until at least 8 hours after TCA in order to avoid

any possible interference with the study (2).

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• After the initial 8-hour post-TCA treatment, buprenorphine 0.01 mg/kg SC should be administered

every 12 hours, in conjunction with meloxicam 0.5 mg/kg SC every 24hours, until the ocular lesions

resolve and no clinical signs of pain and distress are present. Sustained-release preparations of

analgesics are available that could be considered to decrease the frequency of analgesic dosing.

• “Rescue” analgesia should be given immediately after TCA if pre-emptive analgesia and topical

anesthesia are inadequate. If an animal shows signs of pain and distress during the study, a

“rescue” dose of buprenorphine 0.03 mg/kg SC would be given immediately and repeated as often

as every 8 hours, if necessary, instead of 0.01 mg/kg SC every 12 hours. Meloxicam 0.5 mg/kg

SC would be administered every 24 hours in conjunction with the “rescue” dose of buprenorphine,

but not until at least 8 hours post-TCA.

Application of the test chemical

12. The test chemical should be placed in the conjunctival sac of one eye of each animal after gently

pulling the lower lid away from the eyeball. The lids are then gently held together for a few seconds in order

to prevent loss of the material. The other eye, which remains untreated, serves as a control.

Irrigation

13. The eyes of the test animals should not be washed for at least 24 hours following instillation of the

test chemical, except for solids (see paragraph 16), and in case of immediate serious eye damage or eye

irritation effects. At 24 hours a washout may be used if considered appropriate.

14. Use of a satellite group of animals to investigate the influence of washing is not recommended

unless it is scientifically justified. If a satellite group is needed, two rabbits should be used. Conditions of

washing should be carefully documented, e.g., time of washing; composition and temperature of wash

solution; duration, volume, and velocity of application.

Dose level

(1) Testing of liquids

15. For testing liquids, a dose of 0.1 mL is used. Pump sprays should not be used for instilling the test

chemical directly into the eye. The liquid spray should be expelled and collected in a container prior to

instilling 0.1 mL into the eye.

(2) Testing of solids

When testing solids, pastes, and particulate substances, the amount used should have a volume of 0.1mL

or a weight of not more than 100 mg. The test material should be ground to a fine dust. The weight of the

0.1 mL volume of solid material should be recorded after gently compacting it, e.g. by tapping the

measuring container. The amount used should not exceed 100 mg. Application of powdered solid test

chemicals to the eyes will increase blinking and this may contribute to a higher degree of irritation.

16. If the solid test chemical has not been removed from the eye of the test animal by physiological

mechanisms at the first observation time point of 1 hour after treatment, the eye may be rinsed with saline

or distilled water.

(3) Testing of aerosols

17. It is recommended that all pump sprays and aerosols be collected prior to instillation into the eye.

The one exception is for test chemicals in pressurised aerosol containers, which cannot be collected due

to vaporisation. In such cases, the eye should be held open, and the test chemical is administered to the

eye in a simple burst of about one second, from a distance of 10 cm directly in front of the eye. This

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distance may vary depending on the pressure of the spray and its contents. Care should be taken not to

damage the eye from the pressure of the spray. In appropriate cases, there may be a need to evaluate the

potential for “mechanical” damage to the eye from the force of the spray.

18. An estimate of the dose from an aerosol can be made by simulating the test as follows: the test

chemical is sprayed on to weighing paper through an opening the size of a rabbit eye placed directly before

the paper. The weight increase of the paper is used to approximate the amount sprayed into the eye. For

volatile test chemicals, the dose may be estimated by weighing a receiving container before and after

removal of the test material.

Initial test (in vivo serious eye damage/eye irritation test using one animal)

19. It is strongly recommended that the in vivo test be performed initially using one animal.

Observations should allow for determination of severity and reversibility before proceeding to a

confirmatory test in a second animal.

20. If the results of this test indicate the test chemical to be seriously damaging to the eye using the

procedure described, further testing for ocular irritancy should not be performed. Termination of the testing

after a first animal showing serious eye damage is for animal welfare reasons and the rate of

misclassification due to the variability of the animal’s response could be considered.

Confirmatory test (in vivo eye irritation test with additional animals)

21. If a serious eye damage/irreversible effect is not observed in the initial test, the irritant or negative

response should be confirmed using up to two additional animals. If an irritant effect is observed in the

initial test, it is recommended that the confirmatory test be conducted in a sequential manner in one animal

at a time, rather than exposing the two additional animals simultaneously. If the second animal reveals

serious eye damage/irreversible effects, the test is not continued. If results from the second animal are

sufficient to allow for a hazard classification determination, then no further testing should be conducted.

Observation period

22. The duration of the observation period should be sufficient to evaluate fully the magnitude and

reversibility of the effects observed. However, the experiment should be terminated at any time that the

animal shows signs of severe pain or distress (11) (12). To determine reversibility of effects, the animals

should be observed normally for 21 days post administration of the test chemical. If reversibility is seen

before 21 days, the experiment should be terminated at that time.

Clinical observations and grading of eye reactions

23. The eyes should be comprehensively evaluated for the presence or absence of ocular lesions one

hour post-TCA, followed by at least daily evaluations. Animals should be evaluated several times daily for

the first 3 days to ensure that termination decisions are made in a timely manner. Test animals should be

routinely evaluated for the entire duration of the study for clinical signs of pain and/or distress (e.g. repeated

pawing or rubbing of the eye, excessive blinking, excessive tearing) (11) (12) at least twice daily, with a

minimum of 6 hours between observations, or more often if necessary. This is necessary to (i) adequately

assess animals for evidence of pain and distress in order to make informed decisions on the need to

increase the dosage of analgesics and (ii) assess animals for evidence of established humane endpoints

in order to make informed decisions on whether it is appropriate to humanely euthanize animals, and to

ensure that such decisions are made in a timely manner. Fluorescein staining should be routinely used

and a slit lamp biomicroscope used when considered appropriate (e.g., assessing depth of injury when

corneal ulceration is present) as an aid in the detection and measurement of ocular damage, and to

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evaluate if established endpoint criteria for humane euthanasia have been met. Digital photographs of

observed lesions may be collected for reference and to provide a permanent record of the extent of ocular

damage. Animals should be kept on test no longer than necessary once definitive information has been

obtained. Animals showing severe pain or distress should be humanely killed without delay, and the test

chemical assessed accordingly without performing further testing with more animals.

24. Animals with the following eye lesions post-instillation should be humanely killed (refer to Table 1

for a description of lesion grades): corneal perforation or significant corneal ulceration including

staphyloma; blood in the anterior chamber of the eye; grade 4 corneal opacity; absence of a light reflex

(iridial response grade 2) which persists for 72 hours; ulceration of the conjunctival membrane; necrosis of

the conjunctivae or nictitating membrane; or sloughing. This is because such lesions generally are not

reversible. Furthermore, it is recommended that the following ocular lesions be used as humane endpoints

to terminate studies before the end of the scheduled 21-day observation period. These lesions are

considered predictive of serious eye damage and injuries that are not expected to fully reverse by the end

of the 21-day observation period: severe depth of injury (e.g., corneal ulceration extending beyond the

superficial layers of the stroma), limbus destruction >50% (as evidenced by blanching of the conjunctival

tissue), and severe eye infection (purulent discharge). A combination of: vascularization of the cornea

surface (i.e., pannus); area of fluorescein staining not diminishing over time based on daily assessment;

and/or lack of re-epithelialization 5 days after test chemical application could also be considered as

potentially useful criteria to influence the clinical decision on early study termination. However, these

findings individually are insufficient to justify early study termination. Once serious eye damage/irreversible

effects have been identified, an attending or qualified laboratory animal veterinarian or personnel trained

to identify the clinical lesions should be consulted for a clinical examination to determine if the combination

of these effects warrants early study termination. The grades of ocular reaction (conjunctivae, cornea and

iris) should be obtained and recorded at 1, 24, 48, and 72 hours following test chemical application (Table

1). The study should be terminated not earlier than 3 days post instillation if the animals do not develop

ocular lesions. Animals with ocular lesions that are not severe should be observed until the lesions clear,

or for 21 days, at which time the study is terminated. Observations should be performed and recorded at

a minimum of 1 hour, 24 hours, 48 hours, 72 hours, 7 days, 14 days, and 21 days in order to determine

the status of the lesions, and their reversibility or irreversibility. More frequent observations should be

performed if necessary in order to determine whether the test animal should be euthanized out of humane

considerations or removed from the study due to negative results

25. The grades of ocular lesions (Table 1) should be recorded at each examination. Any other lesions

in the eye (e.g. pannus, staining, anterior chamber changes) or adverse systemic effects should also be

reported.

26. Examination of reactions can be facilitated by use of a binocular loupe, hand slit-lamp,

biomicroscope, or other suitable device. After recording the observations at 24 hours, the eyes may be

further examined with the aid of fluorescein.

27. The grading of ocular responses is necessarily subjective. To promote harmonisation of grading

of ocular response and to assist testing laboratories and those involved in making and interpreting the

observations, the personnel performing the observations need to be adequately trained in the scoring

system used.

DATA AND REPORTING

Evaluation of results

28. The ocular irritation scores should be evaluated in conjunction with the nature and severity of

lesions, and their reversibility or lack of reversibility. The individual scores do not represent an absolute

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standard for the irritant properties of a material, as other effects of the test material are also evaluated.

Instead, individual scores should be viewed as reference values and are only meaningful when supported

by a full description and evaluation of all observations.

Test report

29. The test report should include the following information:

Rationale for in vivo testing: weight-of-the-evidence analysis of pre-existing test data, including results from

sequential testing strategy:

‒ description of relevant data available from prior testing;

‒ data derived in each step of testing strategy;

‒ description of in vitro tests performed, including details of procedures, results obtained with test

chemical/reference substances;

‒ description of in vivo dermal irritation / corrosion study performed, including results obtained;

‒ weight-of-the-evidence analysis for performing in vivo study

Test chemical:

‒ identification data (e.g. chemical name and if available CAS number, purity, known impurities,

source, lot number);

‒ physical nature and physicochemical properties (e.g. pH, volatility, solubility, stability, reactivity

with water);

‒ in case of a mixture, components should be identified including identification data of the constituent

substances (e.g. chemical names and if available CAS numbers) and their concentrations;

‒ dose applied;

Vehicle:

‒ identification, concentration (where appropriate), volume used;

‒ justification for choice of vehicle.

Test animals:

‒ species/strain used, rationale for using animals other than albino rabbit;

‒ age of each animal at start of study;

‒ number of animals of each sex in test and control groups (if required);

‒ individual animal weights at start and conclusion of test;

‒ source, housing conditions, diet, etc.

Anaesthetics and analgesics

‒ doses and times when topical anaesthetics and systemic analgesics were administered;

‒ if local anaesthetic is used, identification, purity, type, and potential interaction with test chemical.

Results:

‒ description of method used to score irritation at each observation time (e.g., hand slitlamp,

biomicroscope, fluorescein);

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‒ tabulation of serious eye damage/eye irritation response data for each animal at each observation

time up to removal of each animal from the test;

‒ narrative description of the degree and nature of serious eye damage/eye irritation observed;

‒ description of any other lesions observed in the eye (e.g., vascularization, pannus formation,

adhesions, staining);

‒ description of non-ocular local and systemic adverse effects, record of clinical signs of pain and

distress, digital photographs, and histopathological findings, if any.

‒ If applicable, relevant UNGHS classification (13) for the test chemicals, including not classified.

Interpretation of the results

30. Extrapolation of the results of eye irritation studies in laboratory animals to humans is valid only to

a limited degree. In many cases the albino rabbit is more sensitive than humans to ocular irritants or

chemicals inducing serious eye damage.

31. Care should be taken in the interpretation of data to exclude irritation resulting from secondary

infection.

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LITERATURE

(1) OECD (2019). Guidance Document on an Integrated Approaches on Testing and Assessment for

Serious Eye Damage and Eye irritation. Series on Testing and Assessment No.263. ENV Publications,

Organisation for Economic Cooperation and Development, Paris.

(2) ICCVAM (2010), ICCVAM Test Method Evaluation Report: Recommendations for Routine Use of

Topical Anesthetics, Systemic Analgesics, and Humane Endpoints to Avoid or Minimize Pain and

Distress in Ocular Safety Testing, NIH Publication No. 10-7514, Research Triangle Park, NC, USA:

National Institute of Environmental Health Sciences. Available: http://iccvam.niehs.nih.gov/

methods/ocutox/OcuAnest-TMER.htm

(3) OECD(2020), Test No. 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, OECD Guidelines for the Testing of Chemicals, Section 4, OECD

Publishing. https://doi.org/10.1787/9789264203846-en

(4) OECD (2018), Test No. 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, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris.

https://doi.org/10.1787/9789264203860-en

(5) OECD (2017), Test No. 460: Fluorescein Leakage Test Method for Identifying Ocular Corrosives

and Severe Irritants. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris.

https://doi.org/10.1787/9789264185401-en

(6) OECD (2020), Test No. 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. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing.

https://doi.org/10.1787/9789264242432-en

(7) OECD (2019), Test No. 492: Reconstructed human Cornea-like Epithelium (RhCE) test method

for identifying chemicals not requiring classification and labelling for eye irritation or serious eye damage.

OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing.

https://doi.org/10.1787/9789264242548-en

(8) OECD (2019), Test No. 494: Vitrigel-Eye Irritancy Test Method for Identifying Chemicals Not

Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage. OECD Guidelines for the

Testing of Chemicals, Section 4, OECD Publishing. https://doi.org/10.1787/9f20068a-en

(9) OECD (2019), Test No. 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. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing.

https://doi.org/10.1787/970e5cd9-en

(10) OECD (2000), Guidance Document on the Recognition, Assessment and Use of Clinical Signs as

Humane Endpoints for Experimental Animals Used in Safety Evaluation. OECD Environmental Health and

Safety Publications, Series on Testing and Assessment No. 19 (http://www.oecd.org/ehs/test/monos.htm).

(11) Wright EM, Marcella KL, Woodson JF. (1985), Animal pain: evaluation and control, Lab Animal,

May/June: 20-36.

(12) National Research Council (NRC) (2009), Recognition and Alleviation of Pain in Laboratory

Animals, Washington, DC: The National Academies Press.

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(13) United nations (UN) (2019). Globally Harmonized System of Classification and Labelling of

Chemicals (GHS). Eight revised edition, UN New York and Geneva, 2019. Available at:

https://unece.org/fileadmin/DAM/trans/danger/publi/ghs/ghs_rev08/ST-SG-AC10-30-Rev8e.pdf

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TABLE 1: GRADING OF OCULAR LESIONS

Cornea Grade

Opacity: degree of density (readings should be taken from most dense area)*

No ulceration or opacity .............................................................................................................. 0

Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details

of iris clearly visible ................................................................................................................... 1

Easily discernible translucent area; details of iris slightly obscured ........................................... 2

Nacrous area; no details of iris visible; size of pupil barely discernible ..................................... 3

Opaque cornea; iris not discernible through the opacity ............................................................. 4

Maximum possible: 4

* The area of corneal opacity should be noted

Iris

Normal ......................................................................................................................................... 0

Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia;

or injection; iris reactive to light (a sluggish reaction is considered to be an effect .................... 1

Hemorrhage, gross destruction, or no reaction to light ............................................................... 2

Maximum possible: 2

Conjunctivae

Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and iris)

Normal ......................................................................................................................................... 0

Some blood vessels hyperaemic (injected) ................................................................................. 1

Diffuse,crimson colour; individual vessels not easily discernible............................................... 2

Diffuse beefy red ......................................................................................................................... 3

Maximum possible: 3

Chemosis

Swelling (refers to lids and/or nictating membranes)

Normal ......................................................................................................................................... 0

Some swelling above normal....................................................................................................... 1

Obvious swelling, with partial eversion of lids ........................................................................... 2

Swelling, with lids about half closed ........................................................................................... 3

Swelling, with lids more than half closed ................................................................................... 4

Maximum possible: 4

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ANNEX

DEFINITIONS

1. Not Classified: Test chemicals that are not classified for eye irritation (UN GHS Category 2) or

serious damage to eye (UN GHS Category 1) according to the UN GHS classification (13). Interchangeable

with “UN GHS No Category”.

2. Eye irritation refers to the production of changes in the eye, which are fully reversible, occurring

after the exposure of the eye to a chemical or mixture; these chemicals are classified as UN GHS Category

2, 2A or 2B (13).

3. Serious eye damage refers to the production of tissue damage in the eye, or serious physical

decay of vision, which is not fully reversible, occurring after exposure of the eye to a chemical or mixture .

These chemicals are classified as GHS Category 1 (13).

4. Weight-of-the-evidence (assessment): The strengths and weaknesses of a collection of

information are used as the basis for a conclusion that may not be evident from the individual data.

5. IATA (Integrated Approach to Testing and Assessment): A structured approach used for hazard

identification (potential), hazard characterisation (potency), and/or safety assessment (potential/potency

and exposure) of a chemical or group of chemicals, which strategically integrates and weights all relevant

data to inform regulatory decision regarding potential hazards, risks, and the need for further targeted and

therefore minimal testing.

6. “Defined Approach (DA): a DA consists of a fixed data interpretation procedure (e.g. statistical,

mathematical models) applied to data (e.g. in silico predictions, in chemico, in vitro data) generated with a

defined set of information sources to derive a prediction.”