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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
data is from experimental reports

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)
Principles of method if other than guideline:
To assess the ocular irritation parameter of the test chemical in accordance with OECD 405
GLP compliance:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Ethylenediamine, salt with phosphoric acid
EC Number:
238-914-9
EC Name:
Ethylenediamine, salt with phosphoric acid
Cas Number:
14852-17-6
Molecular formula:
C2H8N2.xH3O4P
IUPAC Name:
ethane-1,2-diamine; phosphoric acid
Test material form:
solid: particulate/powder
Details on test material:
Name of the test chemical: ethylenediamine, salt with phosphoric acid
Molecular Formula: C2H11N2O4P
Molecular Weight: 158.094 g/mol
Physical State: Solid, White powder

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
-Source : In-bred
-Age : 10 to 12 weeks
-Body weight range : -Source : In-bred
-Age : 10 to 12 weeks
-Body weight range : 1.80kg ±200gg
-Identification : By cage tag and corresponding colour body marking.
-Acclimatization : The healthy rabbits selected for study was acclimatized to standard laboratory condition for one week in experimental room under Veterinary examination.
-Randomization : After acclimatization and Veterinary examination three females were randomly selected.
-Accommodation : Animals were housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
-Diet : Pelleted feed supplied by Pranav agro Industries Ltd., Delhi
-Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22-25 deg.C
- Humidity (%): 40-60%
- Air changes (per hr): 10-15 air changes per hour
- Light and Dark Cycle: illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.

Test system

Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 gm moistened with distilled water
Duration of treatment / exposure:
the eyelids were gently pulled together for about one second to prevent loss of material
Observation period (in vivo):
1, 24, 48 and 72 hours after test substance application
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not washed
SCORING SYSTEM:Scale of weighted scores for grading the severity of ocular lesions developed by Draize et al

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein: hand-slit lamp
Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. After recording the observations at 24 hours, the eyes were further examined with the aid of fluorescein.

Results and discussion

In vivo

Results
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
24/48/72 h
Score:
2.5
Max. score:
110
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
In the initial test, the test chemical produced slight redness and mild discharge after 2 hours of application of test compound. This condition was recovered after 24 hours. Furthermore, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc
In the confirmatory test, the test chemical produced slight redness of conjunctivae after two hour of application of test compound upto 24 hours. However, there was no other clinical sign recorded in both of the animals during the whole observation period for 21 days.
Other effects:
The test compound when applied in conjunctival sac of rabbits did not show any observable clinical signs such as cage side activity and pain or stress etc. throughout the observation period of 21 days

Any other information on results incl. tables

TABLE- 1     GRADING OF OCULAR LESIONS

 

S.NO/

SEX

 

OBSERVATION

Score

Total

Total Score

1/F

 

1 hour

24hours

48 hours

72 hours

Cornea

A.       Opacity-Degree of Density

0

0

0

0

0

0×0×5=0

B.       Area of Cornea Involved

0

0

0

0

0

Iris

A.       Values

0

0

0

0

0

0

Conjunctivae

A.       Redness

1

0

0

0

0

1+0+1×5=10

B.       Chemosis

0

0

0

0

0

C.       Discharge 

0

1

0

0

0

2/F

Cornea

A.       Opacity-Degree of Density

0

0

0

0

0

0×0×5=0

B.       Area of Cornea Involved

0

0

0

0

0

Iris

A.       Values

0

0

0

0

0

0

Conjunctivae

A.       Redness

1

0

0

0

0

1+0+1×5=10

B.       Chemosis

0

0

0

0

0

C.       Discharge 

0

1

0

0

0

3/F

Cornea

A.       Opacity-Degree of Density

0

0

0

0

0

0×0×5=0

B.       Area of Cornea Involved

0

0

0

0

0

Iris

A.       Values

0

0

0

0

0

0

Conjunctivae

A.       Redness

1

0

0

0

0

1+0+1×5=10

B.       Chemosis

0

0

0

0

0

C.       Discharge 

0

1

0

0

0

Grand total

30

Mean

10

Eye Irritation Scoring index

2.5

Applicant's summary and conclusion

Interpretation of results:
other: not irritating
Conclusions:
In the initial test, the test chemical produced slight redness and mild discharge after 2 hours of application of test compound. This condition was recovered after 24 hours. Furthermore, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc
In the confirmatory test, the test chemical produced slight redness of conjunctivae after two hour of application of test compound upto 24 hours. However, there was no other clinical sign recorded in both of the animals during the whole observation period for 21 days.
Also the overall irritation score was calculated to be 2.5/110. According to the irritation parameter propised by Kay & Calandra, the test chemical can be consdiered to be practically not irritating to eyes,
It can be classified under the category "Not Classified".
Executive summary:

The acute eye irritation study of the test chemical was conducted in New Zealand White Rabbit as per the OECD- 405 Guideline. 3 female young adult New Zealand White rabbits were used for the study.

One healthy rabbits of body weight 1.96kg was selected for study after acclimatization. Both eyes of rabbits were examined for any abnormal discharge such as eye irritation, ocular defects or pre-existing corneal injury from eye 24 hours prior to application of test compound.

The test compound was applied in the conjunctival sac of rabbit after gently pulling the lower lid away from the eyeball at the dose rate of0.1gm.The lids were then gently held for about one second in order to prevent loss of the material. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hoursafter the treatment. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animal was observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weight before and during the study was observed.

The test chemical produced slight redness and mild discharge after 2 hours of application of test compound. This condition was recovered after 24 hours. Furthermore, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc

A confirmatory test was performed using two additional animals according to the same procedure. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hours after the treatment. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animals were observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weight before and during the study was observed.

In the confirmatory test, the test chemical produced slight redness of conjunctivae after two hour of application of test compound upto 24 hours. However, there was no other clinical sign recorded in both of the animals during the whole observation period for 21 days.

Also the overall irritation score was calculated to be 2.5/110. According to the irritation parameter propised by Kay & Calandra, the test chemical can be consdiered to be practically not irritating to eyes,

It can be classified under the category "Not Classified".