Cyclohexanecarboxylic acid, 2,2,6-trimethyl-, 2-propen-1-yl ester, (1R,6S)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 14 July 2015 and 31 July 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries, 2000

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

NACET10419

IUPAC Name:

NACET10419

Constituent 2

Reference substance name:

(6S)-Allyl 2,2,6-trimethylcyclohexanecarboxylate

IUPAC Name:

(6S)-Allyl 2,2,6-trimethylcyclohexanecarboxylate

Constituent 3

Reference substance name:

1450841-11-8 (planar)

IUPAC Name:

1450841-11-8 (planar)

Test material form:

other: liquid

Details on test material:

Identification: NACET10419
Common/Commercial name: NACET10419
Chemical name: (6S)-Allyl 2,2,6-trimethylcyclohexanecarboxylate
CAS number: 1450841-11-8 (planar)
EC number: not allocated
Batch: KU31213
Purity: 100%
Physical state / Appearance: clear colorless liquid
Expiry date: 20 December 2015
Storage Conditions: room temperature in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.55 or 2.61 kg and were twelve to twenty weeks old. After an acclimatisation period of at least five days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

A volume of 0.1 mL of the test item was placed into the conjunctival sac of the right eye

Duration of treatment / exposure:

Up to 1 hour

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

2

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.

Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.

A volume of 0.1 mL of the test item was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1.

Eight hours after test item application, a subcutaneous injection of post dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.

After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (Draize, J.H, 1977) given in Appendix 2.

Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Any clinical signs of toxicity, if present, were also recorded.

An additional observation was made in one treated eye on Day 7 to assess the reversibility of the ocular effects.

Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 74505 Female

Time point:

other: Highest score at 1, 24, 48, 72 hours and 7days

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal: 74581 Female

Time point:

other: Highest score at 1, 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Results 3

Irritation parameter:

iris score

Basis:

animal: 74505 Female

Time point:

other: Highest score at 1, 24, 48, 72 hours and 7 days

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Results 4

Irritation parameter:

iris score

Basis:

animal: 74581 Female

Time point:

other: Highest score at 1, 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Results 5

Irritation parameter:

other: redness

Basis:

animal: 74505 Female

Time point:

other: Highest score at 1, 24, 48, 72 hours and 7 days

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

other: redness

Basis:

animal: 74581 Female

Time point:

other: Highest score at 1, 24, 48 and 72 hours

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 7

Irritation parameter:

chemosis score

Basis:

animal: 74505 Female

Time point:

other: Highest score at 1, 24, 48, 72 hours and 7 days

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 8

Irritation parameter:

chemosis score

Basis:

animal: 74581 Female

Time point:

other: Highets score at 1, 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

Individual and group mean scores for ocular irritation are given in Table 1 and Table 2. Individual mean scores for ocular irritation required for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and the Globally Harmonized System of Classification and Labelling of Chemicals are given in Table 3.

No corneal or iridial effects were noted during the study.

Moderate conjunctival irritation was noted in both treated eyes 1 and 24 hours after treatment. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 48 Hour observation. Minimal conjunctival irritation was noted in one treated eye at the 72 Hour observation.

One treated eye appeared normal at the 72 Hour observation and the other treated eye appeared normal at the 7 Day observation.

Other effects:

Body weight
Individual body weights and body weight change are given in Table 4.
Both animals showed expected gain in body weight during the study.

Any other information on results incl. tables

Interpretation of Results

The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of cornea involved). For each tissue the score was calculated as follows:

 

Score for conjunctivae      =      (A + B + C) x 2

Score for iris                     =      D x 5

Score for cornea               =      (E x F) x 5

 

Using the numerical data obtained a modified version of the system described by Kay J.H. and Calandra J.C. (1962) (see Appendix 3) was used to classify the ocular irritancy potential of the test item. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test item.

 

If evidence of irreversible ocular damage is noted, the test item will be classified as corrosive to the eye.

 

The results were also interpreted according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and the Globally Harmonized System of Classification and Labelling of Chemicals.

 

Table1     Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	74505Female					74581Female
	IPR= 0					IPR = 0
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	1 Hour	24 Hours	48 Hours	72 Hours
CORNEA
E = Degree of Opacity	0	0	0	0	0	0	0	0	0
F = Area of Cornea Involved	0	0	0	0	0	0	0	0	0
Score (E x F) x 5	0	0	0	0	0	0	0	0	0
IRIS
D	0	0	0	0	0	0	0	0	0
Score (D x 5)	0	0	0	0	0	0	0	0	0
CONJUNCTIVAE
A = Redness	2	2	2	1	0	2	2	2	0
B = Chemosis	2	1	1	0	0	2	1	1	0
C = Discharge	2	1	1	0	0	2	1	0	0
Score (A + B + C) x 2	12	8	8	2	0	12	8	6	0
Total Score	12	8	8	2	0	12	8	6	0

IPR=Initial pain reaction

Table 2     Individual Total Scores and Group Mean Scores for Ocular Irritation

Rabbit Number and Sex	Individual Total Scores At:
	1 Hour	24 Hours	48 Hours	72 Hours	72 Hours
74505Female	12	8	8	2	0
74581Female	12	8	6	0	-
Group Total	24	16	14	2	0
Group Mean Score	12.0	8.0	7.0	1.0	0.0

- =  Observation not required - considered to be zero for calculation of Group Mean Score

Table 3     Individual and Mean Scores for Cornea, Iris and Conjunctivae- According to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and the Globally Harmonized System of Classification and Labelling of Chemicals

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iridial Inflammation	Conjunctival Redness	Conjunctival Chemosis
74505 Female	24 Hours	0	0	1	1
	48 Hours	0	0	1	0
	72 Hours	0	0	0	0
Total		0	0	2	1
Mean		0.0	0.0	0.7	0.3
74581 Female	24 Hours	0	0	2	1
	48 Hours	0	0	1	0
	72 Hours	0	0	0	0
Total		0	0	3	1
Mean		0.0	0.0	1.0	0.3

Table 4     Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
74505 Female	Day 0	Day 7	0.14
	2.61	2.75
74581 Female	Day 0	Day 3	0.04
	2.55	2.59

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

The test item produced a maximum group mean score of 12.0 and was classified as a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and the Globally Harmonized System of Classification and Labelling of Chemicals.

Executive summary:

Introduction

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit.

Results

A single application of the test item to the non-irrigated eye of two rabbits produced moderate conjunctival irritation. One treated eye appeared normal at the 72‑Hour observation and the other treated eye appeared normal at the 7‑Day observation.

Conclusion

The test item produced a maximum group mean score of 12. 0and was classified as a mild irritant (Class4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

 

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures and the Globally Harmonized System of Classification and Labelling of Chemicals.