Diammonium tetrachlorozincate(2-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

version 09 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Diammonium tetrachlorozincate(2-)

EC Number:

238-687-6

EC Name:

Diammonium tetrachlorozincate(2-)

Cas Number:

14639-97-5

Molecular formula:

Cl4Zn.2H4N

IUPAC Name:

diammonium tetrachlorozincdiuide

Details on test material:

The substance was tested as powder.

Test system

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 30mg

Duration of treatment / exposure:

10 seconds

Number of animals or in vitro replicates:

not applicable

Details on study design:

treatments:
-test substance treated chicken eye: treated with 30 mg diammonium tetrachlorozincate
-positive control chicken eye: treated with 30 mg imidazole
-negative control eye: treated with 30µL physiological saline (0.9% (w/v) NaCl solution

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cornea surface was rinsed thoroughly with 20ml physiological saline
- Time after start of exposure: after 10'' of exposure

SCORING SYSTEM:
The control eye and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse.
Minor variations within ±5 minutes were considered acceptable.

The cornea thickness and cornea opacity were measured at all time points.
Fluorescein retention was measured on two occasions, at base line (t=0) and 30 minutes after the post-treatment rinse.


TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope/fluorescein

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

percent corneal swelling

Run / experiment:

up to 75 minutes

Value:

4.3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

percent corneal swelling

Run / experiment:

up to 240 minutes

Value:

11.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: slight indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Value:

3.3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

fluorescein retention score

Value:

3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Any other information on results incl. tables

The mean values of the treated eyes for maximum corneal thickness change, corneal opacity and fluorescein retention are given below. The conclusion on eye irritancy was based on the OECD guideline quantitative assessments.

 

Observation	Value	ICE Class
Mean maximum corneal swelling at up to 75 min	4.3 %	I
Mean maximum corneal swelling at up to 240 min	11.2 %	II
Mean maximum corneal opacity	3.33	IV
Mean fluorescein retention	3.00	IV
Other Observations	Test item was stuck on all cornea surfaces after the post-treatment rinse. The cornea surfaces (3/3) were cleared at 30 minutes after the post-treatment rinse.
Overall ICE Class	1xII 2xIV

Based on this in vitro eye irritation in the isolated chicken eyes test with diammonium tetrachlorozincate(2-),the test item is irritant,UN GHS Classification: Category 1.

 

Positive Control

 

Observation	Value	ICE Class
Mean maximum corneal swelling at up to 75 min	9.6 %	II
Mean maximum corneal swelling at up to 240 min	24.1 %	II
Mean maximum corneal opacity	4.00	IV
Mean fluorescein retention	3.00	IV
Other Observations	Imidazole was stuck on all cornea surfaces after the post-treatment rinse. The cornea surfaces (3/3) were not cleared at 240 minutes after the post-treatment rinse.
Overall ICE Class	1xIII 2xIV

The positive controlImidazole was classified as severely irritating, UN GHS Classification: Category 1.

 

NEGATIVE Control

 

 

Observation	Value	ICE Class
Mean maximum corneal swelling at up to 75 min	0.0 %	I
Mean maximum corneal swelling at up to 240 min	0.0 %	I
Mean maximum corneal opacity	0.00	I
Mean fluorescein retention	0.00	I
Other Observations	None
Overall ICE Class	3xI

 

The negative control Physiological saline was classified as non-irritating, UN GHS Classification: No Category.

Table:Assessment of the general IN VITRO eye irritancy and regulatory GHS classification.

 

UN GHS Classification	Combinations of the three ICE Classes
No Category	3×I 2×I, 1×II
No prediction can be made	Other combinations
Category 1	3×IV 2×IV, 1×III 2×IV, 1×II^* 2×IV, 1×I^* Corneal opacity ≥ 3 at 30 min (in at least 2 eyes) Corneal opacity = 4 at any time point (in at least 2 eyes) Severe loosening of epithelium (in at least 1 eye)

Remark:^*:combinations of categories less likely to occur

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

Based on this in vitro eye irritation assay in isolated chicken eyes with diammonium tetrachlorozincate(2-), the test item is irritant, UN GHS Classification: Category 1.

Executive summary:

An in vitro eye irritation study of the test item was performed in isolated chicken’s eyes. The irritation effects of the test item were evaluated according to the OECD No. 438 guideline (09 October 2017).

 

After the zero reference measurements, the eye was held in horizontal position and 30 mg powdered test itemwas applied onto the centre of the cornea in such a way that the entire surface of the cornea was covered. After 10 seconds, the surface was rinsed with physiological saline. Positive control eyes were treated with 30 mg powdered Imidazole. The negative control eye was treated with 30 µL ofphysiological saline(0.9% (w/v) NaCl solution). In the study, three test item treated eyes, three positive control treated eyes and one negative control treated eye were examined.

The results from all eyes used in the study met the quality control standards. The negative control and positive control results were within the historical control data range in the experiment. Thus, the experiment was considered to be valid.

Slight corneal swelling was observed during the four-hour observation period on test item treated eyes. Severe cornea opacity change (severity 2 on one eye and severity 4 on two eyes) was observed on three eyes. Severe fluorescein retention change (severity 3) was noted on all three eyes.Test item was stuck on all cornea surfaces after the post-treatment rinse. The cornea surfaces were cleared at 30 minutes after the post-treatment rinse.

Based on this in vitro eye irritation assay in isolated chicken eyes with diammonium tetrachlorozincate(2-),the test item is irritant,UN GHS Classification: Category 1.