Poly(oxy-1,2-ethanediyl), .alpha.-[3-[(carboxymethyl)methylamino]-2-hydroxypropyl]-.omega.-(4-nonylphenoxy)-, sodium salt (1:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 03 December 2012 and 03 January 2013

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:

2013

Report date:

2013

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

disodium 2-({2-hydroxy-3-[2-(4-nonylphenoxy)ethoxy]propyl}(methyl)amino)acetate 2-{[3-({1-chloro-3-[2-(4-nonylphenoxy)ethoxy]propan-2-yl}oxy)-2-hydroxypropyl](methyl)amino}acetate

EC Number:

616-248-3

Cas Number:

75627-31-5

Molecular formula:

UVCB not applicable

IUPAC Name:

disodium 2-({2-hydroxy-3-[2-(4-nonylphenoxy)ethoxy]propyl}(methyl)amino)acetate 2-{[3-({1-chloro-3-[2-(4-nonylphenoxy)ethoxy]propan-2-yl}oxy)-2-hydroxypropyl](methyl)amino}acetate

Test material form:

not specified

Details on test material:

Information as provided by the Sponsor. A Certificate of Analysis supplied by the Sponsor

Identification: Nonylphenol ethoxylate, sarcosine derivative
CAS no. 75627-31-5
Batch: 120229AAH
Purity: 81.7%
Expiry Date: 28 February 2017
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 2.38 or 2.58 kg and were twelve to twenty weeks old. After an acclimatization 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. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. 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:

1 hour

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

2 animals were tested in total. (After consideration of the ocular responses produced in the first treated animal, one additional animals were treated.)

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 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.

After consideration of the ocular responses produced in the first treated animal, a second animal was 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 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: 72753 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal: 72817 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 3

Irritation parameter:

iris score

Basis:

animal: 72753 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

iris score

Basis:

animal: 72817 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

other: redness

Basis:

animal: 72753 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 6

Irritation parameter:

other: redness

Basis:

animal: 72817 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 7

Irritation parameter:

chemosis score

Basis:

animal: 72753 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Alopecia around treated eye

Results 8

Irritation parameter:

chemosis score

Basis:

animal: 72817 Male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7 and 14 days

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

Individual and group mean scores for ocular irritation are given in Table 1 and Table 2.

Diffuse corneal opacity was noted in both treated eyes one hour after treatment. Translucent corneal opacity was noted in one treated eye with diffuse corneal opacity noted in the other treated eye at the 24, 48 and 72-Hour observations.

Iridial inflammation was noted in both treated eyes one hour after treatment and at the 24, 48 and 72 Hour observations.

Moderate conjunctival irritation was noted in both treated eyes one hour after treatment and at the 24 and 48 Hour observations. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in the other treated eye at the 72 Hour observation. Minimal conjunctival irritation was noted in both treated eyes at the 7 Day observation.

Alopecia around the treated eye was noted in one animal at the 24, 48, 72 Hour and 7 Day observations.

Both treated eyes appeared normal at the 14-Day observation.

Other effects:

Body weight
Individual body weights and body weight changes are given in Table 3.

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.

Table 1     Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	72753Male						72817Male
	IPR= 2						IPR = 2
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	14 Days		1 Hour	24 Hours	48 Hours	72 Hours	7 Days	14 Days
CORNEA
E = Degree of Opacity	1	2	2	2	0	0		1	1	1	1	0	0
F = Area of Cornea Involved	2	3	3	2	0	0		1	1	1	1	0	0
Score (E x F) x 5	10	30	30	20	0	0		5	5	5	5	0	0
IRIS
D	1	1	1	1	0	0		1	1	1	1	0	0
Score (D x 5)	5	5	5	5	0	0		5	5	5	5	0	0
CONJUNCTIVAE
A = Redness	2	2	2	2	1	0		2	2	1	1	1	0
B = Chemosis	2	2	2	2	1	0		2	2	1	1	1	0
C = Discharge	3	1Al	1Al	1Al	0Al	0		3	3	2	1	0	0
Score (A + B + C) x 2	14	10	10	10	4	0		14	14	8	6	4	0
Total Score	29	45	45	35	4	0		24	24	18	16	4	0

IPR=Initial pain reaction

Al =        Alopecia around treated eye

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	7 Days	14 Days
72753Male	29	45	45	35	4	0
72817Male	24	24	18	16	4	0
Group Total	53	69	63	51	8	0
Group Mean Score	26.5	34.5	31.5	25.5	4.0	0.0

Table 3     Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 14
72753Male	2.38	2.66	0.28
72817Male	2.58	2.97	0.39

Applicant's summary and conclusion

Interpretation of results:

other: The test item produced a maximum group mean score of 34.5 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale)

Remarks:

Criteria used for interpretation of results: other: modified Kay and Calandra classification system

Conclusions:

The test item produced a maximum group mean score of 34.5 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

The test item was also classified as Irritating to eyes (Category 2) according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures. The Signal Word “Warning” and Hazard Statement “H319: Causes serious eye irritation” are therefore required.

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 diffuse to translucent corneal opacity, iridial inflammation and moderate conjunctival irritation. Alopecia around the treated eye was also noted in one animal. Both treated eyes appeared normal at the 14‑Day observation.

 

 

Conclusion

The test item produced a maximum group mean score of 34.5 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

 

The test item was also classified as Irritating to eyes (Category 2) according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures. The Signal Word “Warning” and Hazard Statement “H319: Causes serious eye irritation” are therefore required.