Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: in accordance with guidelines

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): ALGIFERT K+
- Substance type: Dried alkaline extract of marine algae Ascophyllum nodosum dissolved in aqueous solution
- Physical state: Solid, black, micro- flakes, characteristic odour
- Analytical purity: not applicable
- Lot/batch No.: 32325-020509
- Expiry date: 02 May 2012
- Supplier: Valagro S.p.A. (sponsor)
- Storage condition of test material: the Test Article was stored in its original container, as supplied by the Sponsor at room temperature in the Test Article Stores of INTOX. The stability of the test substance in storage will be the responsibility of the Sponsor.
- pH: 9.2 (1% aqueous solution)

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Serum Institute of India Ltd., Pune; a supplier approved by the test facility management.
- Age at study initiation: Young adults (6 to 7 months)
- Weight at study initiation: 2.58 to 2.65 kg
- Housing: Animals were housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
- Diet (e.g. ad libitum): 'Amrut' brand extruded pelleted rabbit feed manufactured by Pranav Agro Ltd., Pune, provided ad libitum
- Water (e.g. ad libitum): Potable water, passed through Aqua guard water filter, and subjected to ultra violet irradiation, provided ad libitum in stainless steel bowls, throughout the acclimation and study period.
- Acclimation period: The animals were acclimated for eleven days in the experimental room before start of the experiment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23oC
- Humidity (%): 30-70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark.

IN-LIFE DATES: From: 15 September 2009 To: 23 September 2009

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated right eye served as a control
Amount / concentration applied:
100 mg
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
1, 24, 48, 72 hours and day 7 after the application.
Number of animals or in vitro replicates:
three males
Details on study design:
The test article ALGIFERT K+, was pulverised by using mortar and pestle before instillation in the eye of the rabbit. On test day the rabbits were fixed in rabbit-holder during application. The test article, in the amount of 100 mg, was placed in the conjunctival sac of left eye of each animal after gently pulling the lower lid away from the eyeball. Since a volume of 0.1 ml of the test article, gently compacted in a measuring cylinder, was found to weight 116 mg i.e. exceding 100 mg, the test dose was administered as 100 mg only. The eyelids were then gently held together for about one second to prevent loss of test article. The right eye remained untreated and served as the reference control. The treated and untreated eyes were washed at 1 hour following instillation with Millipore Elix- 10 system analytical grade water, to remove any residual test article.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 h
Score:
6.7
Max. score:
8
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 24 hs.
Score:
3.3
Max. score:
4
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 48 hs.
Score:
1.3
Max. score:
2
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 72 hs.
Score:
0.7
Max. score:
2
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 7 days
Score:
0
Max. score:
0
Irritant / corrosive response data:
Maximum Group Mean Score: 6.7
Persistence of Score: Group mean total score at 48 hours >0 (as per Modified Version of the Kay and Calandra System)

Any other information on results incl. tables

SUMMARY OF OCULAR IRRITANCY SCORES

Animal ID & Sex

Sum of Irritancy Scores at

1

24

48

72

7

h

h

h

h

Day

RB7834  Male

4

2

0

0

-

RB7835  Male

8

4

2

0

-

RB7836  Male

8

4

2

2

0

Group mean of the total score

6.7

3.3

1.3

0.7

0.0

Maximum Group Mean Score

6.7

Classification as per Modified Version of the Kay and Calandra System

Persistence of Score

Group mean total score at 48 hours >0

Descriptive Rating

Mild-irritant

Individual Animal Ocular Irritation DATA

Animal ID & Sex

Findings

1

24

48

72

7

 

h

h

h

h

Day

 

RB7834   Male

Cornea

( A ) opacity - degree of density

0

0

0

0

-

 

( B ) area involved

0

0

0

0

-

 

Score A x B x 5 =

0

0

0

0

-

 

Iris

( C ) function and aspect

0

0

0

0

-

 

Score C x 5 =

0

0

0

0

-

 

Conjunctiva

( D ) redness

1

1

0

0

-

 

( E ) chemosis

1

0

0

0

-

 

( F ) discharge

0

0

0

0

-

 

Score ( D + E + F ) x 2 =

4

2

0

0

-

 

Sum of Scores

4

2

0

0

-

 

RB7835  Male

Cornea

( A ) opacity - degree of density

0

0

0

0

-

 

( B ) area involved

0

0

0

0

-

 

Score A x B x 5 =

0

0

0

0

-

 

Iris

( C ) function and aspect

0

0

0

0

-

 

Score C x 5

0

0

0

0

-

 

Conjunctiva

( D ) redness

2

1

0

0

-

 

( E ) chemosis

2

1

0

0

-

 

( F ) discharge

0

0

0

0

-

 

Score ( D + E + F ) x 2 =

8

4

0

0

-

 

Sum of Scores

8

4

0

0

-

 

RB7836  Male

Cornea

( A ) opacity - degree of density

0

0

0

0

0

 

( B ) area involved

0

0

0

0

0

 

Score A x B x 5 =

0

0

0

0

0

 

Iris

( C ) function and aspect

0

0

0

0

0

 

Score C x 5 =

0

0

0

0

0

 

Conjunctiva

( D ) redness

2

1

1

1

0

 

( E ) chemosis

2

1

0

0

0

 

( F ) discharge

0

0

0

0

0

 

Score ( D + E + F ) x 2 =

8

4

2

2

0

 

Sum of Scores

8

4

2

2

0

 

Group mean of the total score

6.7

3.3

0.7

0.7

0.0

 

Maximum Group Mean Score

6.7

Classification as per Modified Version of the Kay and Calandra System

Persistence of Score

Group mean total score at 48 hours > 0

Descriptive Rating

Mild irritant

Class

4

INDIVIDUAL ANIMAL CLINICAL SIGNS

Animal ID.

Time of Observation : Study Day

1

2

3

4

5

6

7

RB7834

NAD

NAD

NAD

NAD

-

-

-

RB7835

NAD

NAD

NAD

NAD

-

-

-

RB7836

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Day 1 is the day of treatment.   NAD – No abnormality detected,

Applicant's summary and conclusion

Interpretation of results:
other: mild irritant
Remarks:
Criteria used for interpretation of results: other: modified version of the Kay and Calandra System
Conclusions:
Acute eye irritation / corrosion test was performed on ALGIFERT K+ in rabbits, in compliance with the Organization for Economic Cooperation and Development (OECD) Guidelines for Testing of Chemicals, Section 4, N° 405 - Acute Eye Irritation / Corrosion, adopted on 24th April 2002.
This acute eye irritation / corrosion test, comprising of an initial test performed on one rabbit, followed by a confirmatory test on two more rabbits, revealed a maximum irritancy score of 6.7 at 1h post-application, decreasing gradually thereafter to 0 on day 7. The results indicated that the induced changes were reversible in nature, and hence were classified as ‘irritation’ and not as ‘corrosion’.
When examined in light of the criteria for evaluation, i.e. modified version of the Kay and Calandra System, based on the maximum score and persistence of the irritancy score, ALGIFERT K+ is classified as mild irritant to eye of rabbit.
Executive summary:

Testing ofALGIFERT K+for its acute irritation / corrosion potential in the rabbit eye was performed in compliance with the Organization for Economic Cooperation and Development (OECD) Guidelines for Testing of Chemicals, Section 4,405 - Acute Eye Irritation / Corrosion study, adopted 24th April 2002. This test was performed in accordance with the sequential testing strategy recommended in this OECD Test Guideline.Initially the test was performed using one animal. As the observations at this stage did notsuggest the test article to be severely irritating orcorrosive, the study was continued to conduct a confirmatory test involving two more animals.The test article was pulverized and an amount of 100 mg was applied as a single instillation into the lower conjunctival sac of left eye of the rabbits while the untreated right eye served as a control. Eyes were rinsed with water1 hour after application of the test article. The degree of eye irritation / corrosion was evaluated by scoring changes in conjunctiva, cornea and iris, at 1, 24, 48, 72 hours and day 7 after the application. Other effects in the eye and adverse systemic effects, if any, were observed and described. Body weights were recorded on the day of treatment and at termination of the study.The tests performed on rabbits revealed a maximum irritancy score of 6.7 at 1 h post-application, decreasing thereafter to 0 (zero) on day 7, which indicated that the induced changes were reversible in nature, and hence could be classified as ‘irritation’ and not as ‘corrosion’.The treated rabbits did not exhibit any signs of systemic toxicity throughout the period of the study. Their body weight gain was not adversely affected through the study period. When examined in light of the criteria for evaluation, i.e. modified version of the Kay and Calandra System, based on the maximum score and persistence of the irritancy score,ALGIFERT K+is classified as a mild-irritant to eye in rabbit.