Resin acids and Rosin acids, barium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Mar - Apr 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - See attached justification for WoE: Eye irritation

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Reference 2

Reference Type:

other: Amendment No. 1 to the study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability 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:

09 Oct 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Resin acids and Rosin acids, barium salts

EC Number:

269-142-0

EC Name:

Resin acids and Rosin acids, barium salts

Cas Number:

68188-14-7

Molecular formula:

N/A

IUPAC Name:

Resin acids and Rosin acids, barium salts

Test material form:

solid

Details on test material:

Specific details on test material used for the study
SOURCE OF TEST MATERIAL
- Batch number of test material: EXP NR.3757-01
- Expiration date of the lot/batch: 31 Dec 2029
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: refrigerator
- Stability under storage conditions: The stability of the test substance under storage conditions is
guaranteed until 31 Dec 2029 as indicated by the sponsor, and the sponsor holds this responsibility.
- Stability under test conditions: no data

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source: BASF SE Ludwigshafen, Germany
- lot/batch number of test material: EXP NR.3757-01
- Expiration date of the lot/batch: 31 Dec 2029
- Purity: 100 % UVCB
- Physical state: Solid

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Refrigerator
- Stability under storage conditions: was guaranteed over the study period

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Freshly slaughtered cattles, Schlachthof Alzey, Emil Färber GmbH & Co. KG, Robert-Bosch-Strasse 23, 55232 Alzey, Germany
- Characteristics of donor animals (age): minimum 12 months, maximum 60 months
- indication of any existing defects or lesions in ocular tissue samples: Corneas free of defects (opacity, scratches, pigmentation etc.)

Test system

Vehicle:

water

Remarks:

Deionized

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL, NC AND PC
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20% test material / Imidazole (w/v) suspension in deionized water

Duration of treatment / exposure:

4 hours

Number of animals or in vitro replicates:

Each treatment group (test substance, NC and PC) consisted of 3 corneas

Details on study design:

SELECTION AND PREPARATION OF CORNEAS :
Corneas free of defects (opacity, scratches, pigmentation etc.) were dissected with a
2 to 3 mm rim of sclera. Isolated corneas were mounted in cornea holders that consist of anterior and posterior chambers. Both chambers were filled to excess with pre-warmed Eagles’s MEM (without phenol red) and then equilibrated in a vertical position at about 32°C for at least 1 hour. After the equilibration period the medium in both chambers was replaced with fresh pre-warmed medium, and initial corneal opacity readings were taken for each cornea with an opacitometer.

QUALITY CHECK OF THE ISOLATED CORNEAS :
Any corneas that showed macroscopic tissue damage or an opacity value < 553 opacity units2 were discarded. The remaining corneas were then distributed into negative control, positive control and treatment groups.

NUMBER OF REPLICATES : 3

NEGATIVE CONTROL USED : yes

POSITIVE CONTROL USED : yes

APPLICATION DOSE AND EXPOSURE TIME : 750µL, 4 hours

TREATMENT METHOD: open chamber

POST-INCUBATION PERIOD: no

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 3

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: yes,
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry (OD490)
- Others (pertinent visual observations): Before measurement, each cornea was observed visually and observations were recorded. Final corneal opacity readings were taken for each cornea with an opacitometer.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: according to the OECD Guideline 437 (adopted 09 Oct 2017) for evaluation of results.
- IVIS ≤ 3: NO Category
- IVIS > 3; ≤ 55: No prediction can be made
- IVIS > 55 : Category 1
- IVIS 1,5 - 4,5: "borderline" result
- IVIS 45 - 65: "borderline" result
A “borderline“ evaluation was determined statistically using historic BASF data and takes the test facility-specific variance of the test method into account. This evaluation is an amendment to the evaluation provided in OECD Guideline 437.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

8.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: No indication of corrosion

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Any other information on results incl. tables

Table 1: In Vitro Irritancy score (IVIS) of the test substance, the NC and the PC

Test substance identification	Cornea- No.	Opacity per cornea	Permeability per cornea	per cornea	IVIS per group
					mean	SD
	7	0.0	0.191	2.9
19/0421-1	8	0.0	0.462	6.9	8.1	5.8
	9	8.1	0.420	14.4
	1	9.3	0.003	9.3
NC	2	3.4	0.005	3.5	6.8	3.0
	3	7.5	0.008	7.6
	4	84.7	1.451	106.4
PC	5	105.3	2.193	138.2	118.0	17.5
	6	81.4	1.880	109.5

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

The test method according to the regulatory accepted protocol at the time of reporting does not allow for the evaluation of eye irritation. The result does not exclude an irritation potential of the test substance. For final assignment of a risk phrase at present, results from another study would be needed.

Executive summary:

The potential of the test substance to cause ocular irritation or serious damage to the eyes was assessed by a single topical application of 750 µL 20% test-substance preparation to the epithelial surface of isolated bovine corneas.

Three corneas were treated with the test-substance preparation for an exposure period of 4 hours. In addition to the test substance, a negative control (NC; deionized water) and a positive control (PC; 20% imidazole in deionized water) were applied to three corneas each.

Corneal opacity was measured quantitatively as the amount of light transmitted through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were used to calculate an In Vitro Irritancy Score of the test substance.

The mean IVIS (8.1) of the corneas treated with the 20% test-substance preparation does not indicate serious eye damage (BCOP cut-off threshold 55). The results of the test substance, the negative control (deionized water) and the positive control (20% imidazole) fulfill the acceptance criteria and demonstrate the validity of the assay.

Based on the results observed and by applying the evaluation criteria described, it was concluded that the test substance does not cause serious eye damage in the Bovine Corneal Opacity and Permeability Test (BCOP Test) under the test conditions chosen. The test method according to the regulatory accepted protocol at the time of reporting does not allow for the evaluation of eye irritation. The result does not exclude an irritation potential of the test substance. For final assignment of a risk phrase at present, results from another study would be needed.