Reaction product of salicylaldehyde and formaldehyde and phenol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 October 2012 to 24 October 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

yes

Remarks:

(see "Principles of method if other than guideline" for information)

Principles of method if other than guideline:

The Guideline states that the relative humidity should be at least 30% and preferably not exceed 70%, other than during room cleaning, the aim should be 50-60%. The range in the study was reported to be 38 - 82%. This deviation is considered to have no impact on the outcome of the study and interpretation of the results.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of salicylaldehyde and formaldehyde and phenol

EC Number:

632-556-0

Cas Number:

304859-44-7

Molecular formula:

C6H5O(C13H10O2)m(C7H6O)nH values on m and n are variable as this is a UVCB substance

IUPAC Name:

Reaction product of salicylaldehyde and formaldehyde and phenol

Test material form:

solid

Details on test material:

- Description: Light red / solid
- Storage conditions: Room temperature (15-30 °C), protected from humidity

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: ~13 weeks old
- Weight at study initiation: 3157 g
- Housing: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages.
- Diet: ad libitum
- Water: The animals received municipal tap water, as for human consumption, ad libitum, from an automatic system. The quality control analysis is performed once every three months and microbiological assessment is performed monthly
- Acclimation period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3 °C
- Humidity: 38 - 82%
- Air changes: 15 - 20 air exchanges/hour
- Photoperiod: 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 g

Duration of treatment / exposure:

The eyelids were held closed for a few seconds to prevent the loss of the test material.

Observation period (in vivo):

3 weeks

Number of animals or in vitro replicates:

1 male

Details on study design:

TREATMENT
- Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition and any with ocular lesions were rejected.
- An initial test was performed using one animal. The test material was instilled into the conjunctival sac of the left eye. The eyelids were held closed for a few seconds to prevent the loss of the test material. The contra lateral eye served as the control. Immediately after the administration of the test material, an assessment of the initial pain reaction was made according to a six point scale.
- After consideration of the ocular responses produced in the first animal, one hour after the treatment of the first animal, two additional animals were treated.

REMOVAL OF TEST SUBSTANCE
- The treated eye was rinsed with physiological saline solution at 1 hour observation.

OBSERVATIONS
- The eyes were examined at 1, 24, 48, 72 hours, 1, 2 and 3 weeks after treatment. The duration of the observation period was sufficient to identify reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill-health during the study were recorded. At the end of the observation period, the animal was sacrificed by intramuscular injections of CP-Ketamin 10 % and CP-Xylazin 2 % followed by iv. Euthasol® 40 % anaesthesia. Death was verified by checking pupil and corneal reflex and the absence of respiration.
- Measurement of Body Weight: Individual body weight was recorded on the day of treatment and end of the experiment.

SCORING SYSTEM:
- The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (24 April 2002).

CLASSIFICATION
Individual reactions of the animals were recorded at each observation time. The nature, severity and duration of all lesions observed were described.
Results were presented and interpreted according to Regulation (EC) No 1272/2008, as follows:

Irreversible effects on the eye/serious damage to eyes (Category 1)
- Substances that have the potential to seriously damage the eyes are classified in Category 1 (irreversible effects on the eye). These observations include animals with grade 4 cornea lesions and other severe reactions (e.g., destruction of cornea) observed at any time during the test, as well as persistent corneal opacity, discolouration of the cornea by a dye substance, adhesion, pannus, and interference with the function of the iris or other effects that impair sight.
- Category for irreversible eye effects
If, when applied to the eye of an animal, a substance produces:
- at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days; and/or
- at least in 2 of 3 tested animals, a positive response of corneal opacity ≥ 3 and/or iritis > 1.5 calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

Reversible effects on the eye/irritating to eyes (Category 2)
- Substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes).
- Category for reversible eye effects
If, when applied to the eye of an animal, a substance produces:
- at least in 2 of 3 tested animals, a positive response of: corneal opacity ≥ 1 and/or iritis ≥ 1, and/or conjunctival redness ≥ 2 and/or conjunctival oedema (chemosis) ≥ 2 calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

fully reversible within: 3 weeks

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

not fully reversible within: 3 weeks

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

not fully reversible within: 3 weeks

Results 5

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 3 weeks

Irritant / corrosive response data:

EXAMINATION OF EYE IRRITANCY
- The eyes were examined at 1, 24, 48, 72 hours, 1, 2, and 3 weeks after the application.
- Initial Pain Reaction (IPR) (score 1) was observed in the animal.
- One hour after the application: Conjunctival redness (score 2), chemosis (score 2), discharge (score 3) were found in the animal. Corneal opacity (score 1, area 4) was observed in the animal. Test material staining was observed around the eye area.
- At 24, 48 and 72 hours after the application: Conjunctival redness (score 2), chemosis (score 4), discharge (score 3) were found in the animal. Corneal opacity (score 2, area 3) was observed in the animal.
- At 1 week after the application: Conjunctival redness (score 2), chemosis (score 2), discharge (score 3) were found in the animal. Corneal opacity (score 2, area 4) was observed in the animal.
- At 2 weeks after the application: Conjunctival redness (score 1), chemosis (score 1), discharge (score 2) were found in the animal. Corneal opacity (score 2, area 4) was observed in the animal.
- At 3 weeks after the application: Conjunctival redness (score 1), was noted in the animal. Corneal opacity (score 2, area 4) was observed in the animal.
- The animal individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:
chemosis: 4.00
discharge: 3.00
redness: 2.00
cornea opacity: 2.00
iris: 0.00

Other effects:

MORTALITY
- There was no mortality observed during the study.

BODY WEIGHTS
- The body weight and body weight change were considered to be normal with no indication of a treatment related effect.

CLINICAL OBSERVATION
- General daily examination: There were no clinical signs observed that could be related to treatment.

Any other information on results incl. tables

Table 1: Individual Scores for Ocular Irritation

Time	Score of Irritation
	Conjunctivae			Opacity of Cornea		Iris	Control Eye
	R	CH	D	OD	OE	R
1 hour	2	2	3	1	4	0	0
24 hours	2	4	3	2	3	0	0
48 hours	2	4	3	2	3	0	0
72 hours	2	4	3	2	3	0	0
1 week	2	2	3	2	4	0	0
2 weeks	1	1	2	2	4	0	0
3 weeks	1	0	0	2	4	0	0

R = Redness, OD = Opacity degree of density, CH = Chemosis, OE = Extent of opaque area, D = Discharge

Applicant's summary and conclusion

Interpretation of results:

other: EU criteria: Category 1, H318: Causes serious eye irritation

Conclusions:

Under the conditions of this study, the test material caused significant conjunctival and corneal irritant effects at one hour after application and the effects were not fully reversible within 3 weeks. The test material requires classification as an eye irritant (Irreversible effects on the eye/serious damage to eyes (Category 1)).

Executive summary:

The eye irritation potential of the test material was investigated in accordance with the standardised guideline OECD 405, under GLP conditions.

An acute eye irritation study of the test material was performed in New Zealand White rabbits. The test material was placed into the conjunctival sac of the left eye of the single animal. The untreated right eye served as control. An amount of 0.1 g of the test material was administered as a single dose. For ethical reasons, an initial test was performed using a single animal. As this test indicated that the test material is severely irritating or corrosive to the eye, further testing was not be performed.

Individual body weight was recorded on the day of treatment and end of the experiment. Morbidity and clinical signs of toxicity were checked daily. The eyes were examined at 1, 24, 48, 72 hours, 1, 2 and 3 weeks after the application.

Initial Pain Reaction (IPR) (score 1) was observed in the animal. One hour after the application: Conjunctival redness (score 2), chemosis (score 2), discharge (score 3) were found in the animal. Corneal opacity (score 1, area 4) was observed in the animal. Test material staining was observed around the eye area. At 24, 48 and 72 hours after the application: Conjunctival redness (score 2), chemosis (score 4), discharge (score 3) were found in the animal. Corneal opacity (score 2, area 3) was observed in the animal.

At 1 week after the application: Conjunctival redness (score 2), chemosis (score 2), discharge (score 3) were found in the animal. Corneal opacity (score 2, area 4) was observed in the animal. At 2 weeks after the application: Conjunctival redness (score 1), chemosis (score 1), discharge (score 2) were found in the animal. Corneal opacity (score 2, area 4) was observed in the animal. At 3 weeks after the application: Conjunctival redness (score 1), was noted in the animal. Corneal opacity (score 2, area 4) was observed in the animal.

No clinical signs of systemic toxicity were observed in the animal during the study and no mortality occurred. The body weights of the animal were considered to be within the normal range of variability.

The animal individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows: chemosis: 4.00, discharge: 3.00, redness: 2.00, cornea opacity: 2.00 and iris: 0.00.

Under the conditions of this study, the test material caused significant conjunctival and corneal irritant effects at one hour after application and the effects were not fully reversible within 3 weeks. The test material requires classification as an eye irritant (Irreversible effects on the eye/serious damage to eyes (Category 1)).