Bis(5-oxo-L-prolinato-N1,O2)zinc

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 May 2003 to 07 August 2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Bis(5-oxo-L-prolinato-N1,O2)zinc

EC Number:

239-473-5

EC Name:

Bis(5-oxo-L-prolinato-N1,O2)zinc

Cas Number:

15454-75-8

Molecular formula:

C10H12N2O6Zn

IUPAC Name:

(3'aR)-3,3',6,6'-tetraoxo-3a,3'a,4,4',5,5',6,6'-octahydro-3H,3'H-1,1'-spirobi[pyrrolo[1,2-c]1-oxa-3-aza-2-zincacyclopentane]-1,1-bis(ylium)-2,2'-diuide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

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:
- Weight at study initiation: 3.0 - 3.24 kg
- Housing: Bracket cages with 1 animal/cage.
- Diet: Each animal was limited to 150 g feed per day.
- Water: Tap water ad libitum.
- Acclimation period: Upon conducting observations and measurements of the following and monitoring the growth status of the animals during the 11-day quarantine/acclimation period, there were no abnormalities found.
1) Clinical signs: An observation of clinical signs based on visual examinations was carried out every day for all animals, with the exception of holidays.
2) Measurement of body weight: The body weights of all of the animals were measured on the day that the quarantine/acclimation period started and on the day that quarantine/acclimation ended.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 3 °C
- Humidity (%): 50 ± 20 %
- Air changes (per hr): All-fresh ventilation of 10 to 15 cycles per hour.
- Photoperiod (hrs dark / hrs light): 12 hours from 7 am to 7 pm; 200 to 500 lux.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: The rabbits were fixed into place using retention devices. 0.1 g of the test article was ocularly instilled into the right eye after pulling the lower eyelid away from the eyeball into a saclike state. After ocular instillation, the upper and lower eyelids were gently held together for approximately 2 to 3 seconds.

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

Three

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: The eye was washed for approximately 10 seconds using distilled water for injection that was heated to approximately 30 °C.
- Time after start of exposure: 24 hours after ocular instillation.

SCORING SYSTEM:
(1) Cornea:
Opacity: Degree of opacity (reading taken from the most opaque area)
0 - No ulceration or opacity
1 - Scattered or diffuse area of opacity (other than slight dulling of normal luster); details of iris clearly visible
2 - Easily discernible translucent area; details of iris slightly obscured
3 - Nacrous area; no details of iris visible; size of pupil barely discernible
4 - Opaque cornea; iris is not discernible through the opacity
 
(2) Iris:
0 - Normal
1 - Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia; iris reactive to light (a sluggish reaction is considered to be an effect)
2 - Hemorrhage,gross destriction, or no reaction to light
 
(3) Redness of the conjunctivae (refers to palpebral and bulbar conjunctivae; sxcluding cornea and iris strictest manner by comparing them to the reference eye):
0 - Normal
1- Some blood vessels hyperemic( injected)
2 - Diffuse, crimson color; individual vessels not easily discernible
3 - Diffuse, beefy red
 
4) Chemosis: Lids and/or nictating membranes
0 - Normal
1 - Some swelling above normal
2 -Obvious swelling, with partial extroversion of lids
3 - Swelling with lids about half closed
4 - Swelling with lids more than half closed

Ocular irritation potential was also graded in the same manner at 24, 48, and 72 hours after ocular instillation. As irritation was still present 72 hours after ocular instillation, observations were also conducted 1 week, 10 days, 15 days, and 21 days after ocular instillation.

General observation of symptoms
Observations were conducted for all animals once daily, with the exception of holidays.

Measurement of body weight
The body weight of all of the animals was measured when ocularly instilling the test article and during the observations (excluding the observation conducted 1 hour after ocular instillation).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.67

Max. score:

3

Reversibility:

not reversible

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.33

Max. score:

3

Reversibility:

not reversible

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

not reversible

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

not reversible

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

1 hour after ocular instillation of the test material in its original form, cornea opacity to the extent where the details of the iris were clearly visible was observed in 1/3 of the animals. Congestion of the iris was observed in all animals, hyperaemia of the conjunctivae was observed in all of the animals, and swelling with lids about half closed was observed in all of the animals.
24 hours after ocular instillation, cornea opacity to the extent where details of the iris were clearly visible was observed in all of the animals, and congestion of the iris was observed in all animals. Redness of the conjunctivae of a crimson colour was observed in 2/3 of the animals and hyperaemia was observed in 1/3 of the animals; swelling with lids about half closed was observed in 2/3 of the animals and obvious swelling was observed in 1/3 of the animals.
48 hours after ocular instillation, cornea opacity to the extent where the details of the iris were slightly obscured was observed in 2/3 of the animals, and to the extent where the details of the iris were clearly visible was observed in 1/3 of the animals. Congestion of the iris was observed in all of the animals. Redness of the conjunctivae of a beefy red colour was observed in 1/3 of the animals and of a crimson colour was observed in 2/3 of the animals; swelling with lids about half closed was observed in 2/3 of the animals and obvious swelling was observed in 1/3 of the animals.
72 hours after ocular instillation, cornea opacity to the extent where the details of the iris were slightly obscured was observed in 2/3 of the animals, and to the extent where the details of the iris were clearly visible was observed in 1/3 of the animals. Congestion of the iris was observed in all animals; redness of the conjunctivae of a beefy red colour was observed in 2/3 of the animals and of a crimson colour in 1/3 of the animals; swelling with lids about half closed was observed in 1/3 of the animals and obvious swelling in 2/3 of the animals.
Since ocular irritation did not disappear 72 hours after ocular instillation, observations were continued. 1 week after ocular instillation, cornea opacity to the extent where the details of the iris were slightly obscured was observed in 2/3 of the animals, and to the extent where the details of the iris could be clearly discerned in 1/3 of the animals; redness of the conjunctivae of a beefy red colour with haemorrhaging, and some swelling above normal were observed in all animals. 10 days after ocular instillation, cornea opacity to the extent where the details of the iris were clearly visible was observed in 2/3 of the animals, redness of the conjunctivae of a beefy red colour was observed in all animals, and some swelling above normal was observed in 2/3 of the animals. 15 days after ocular instillation cornea opacity to the extent where the details of the iris were clearly visible was observed in 1/3 of the animals, redness of the conjunctivae of a beefy red colour was observed in all animals, and some swelling above normal was observed in 2/3 of the animals. 21 days after ocular instillation, redness of the conjunctivae of a beefy red colour was observed in all animals, and some swelling above normal was observed in 2/3 of the animals.

Any other information on results incl. tables

Eye Irritation Scores of the Test Material

No.	Sex	Reading time	Cornea		Iris		Conjuctiva
			Cornea	Mean score	Iris	Mean score	Redness	Mean score	Chemosis	Mean score	Secreta
00101 00102 00103	M M M	1 hour	2 2 1	1.7	1 1 1	1.0	1 1 1	1.0	3 3 3	3.0	3 3 3
00101 00102 00103	M M M	24 hours	1 1 1	1.0	1 1 1	1.0	2 2 1	1.7	3 3 2	2.7	0 0 0
00101 00102 00103	M M M	48 hours	2 2 1	1.7	1 1 1	1.0	3 2 2	2.3	3 3 2	2.7	3 3 3
00101 00102 00103	M M M	72 hours	2 2 1	1.7	1 1 1	1.0	3 3 2	2.7	2 3 2	2.3	2 2 2
00101 00102 00103	M M M	1 week	2 2 1	1.7	0 0 0	0.0	3 3 3	3.0	1 1 1	1.0	0 0 0
00101 00102 00103	M M M	10 days	1 1 0	0.7	0 0 0	0.0	3 3 3	3.0	1 1 0	0.7	0 1 0
00101 00102 00103	M M M	15 days	1 1 0	0.3	0 0 0	0.0	3 3 3	3.0	1 1 0	0.7	0 0 0
00101 00102 00103	M M M	21 days	0 0 0	0.0	0 0 0	0.0	3 3 3	3.0	1 1 0	0.7	0 0 0

M = Male

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the conditions of this study, since lesions continued even 21 days after exposure, it was concluded that the test material in its original form is classified as a Category 1 eye irritant.

Executive summary:

The eye irritation potential of the test material was investigated in a study which was in accordance with the standardised guideline OECD 405 (Acute Eye Irritation/Corrosion).

Three male white New Zealand rabbits were used in this study. The rabbits were fixed into place using retention devices. 0.1 g of the test article was ocularly instilled into the right eye after pulling the lower eyelid away from the eyeball into a saclike state. After ocular instillation, the upper and lower eyelids were gently held together for approximately 2 to 3 seconds. The left eye remained untreated and served as the reference control. 24 hours after ocular instillation, the eye was washed for approximately 10 seconds using distilled water for injection that was heated to approximately 30 °C. Observations of clinical signs based on visual examinations were carried out every day for all animals, with the exception of holidays. The body weight of all of the animals was measured when ocularly instilling the test article and during the observations (excluding the observation conducted 1 hour after ocular instillation).

Instillation of undiluted test material into the eyes of the rabbits resulted in ocular irritation. Since ocular irritation did not disappear 72 hours after ocular instillation, observations were continued. 21 days after ocular instillation, redness of the conjunctivae of a beefy red colour was observed in all animals, and some swelling above normal was observed in 2/3 of the animals.

Under the conditions of this study, it was concluded that the test material in its original form has a risk of serious damage to eyes.