2,2'-dibromobiphenyl

Endpoint summary

Description of key information

OECD 439: not irritating to skin

OECD 437: not irritating to eye

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Oct 09, 2014 - Jan 29, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

July 26, 2013

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Council Regulation (EC) No. 440/2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006. B.40.bis. In vitro skin corrosion: human skin model test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from a single donor

Justification for test system used:

standard model

Vehicle:

unchanged (no vehicle)

Remarks:

No vehicle used in this study; The test item was applied neat to the tissues.

Details on test system:

CELL CULTURE
- Supplier: EpiSkin/SkinEthic Laboratories, Lyon, France)
- Source: human keratinocytes cultured on a polycarbonate filter in conditions which permit their terminal differentiation
- Format: 24 well plate
- Batch: 14-RHE-024
- Expires: Oct 27, 2014

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment: room temperature
- Temperature of post-treatment incubation: 37°C

REMOVAL OF THE TEST MATERIAL AND CONTROL
At the end of the exposure periods, the test item, positive and negative control was removed immediately by gently rinsing with a minimum volume of 20 mL DPBS using a pipette. Excess DPBS was removed by gently shaking the tissue inserts and blotting the bottom of the tissue inserts with blotting paper.

Control samples:

yes, concurrent negative control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 16 mg
- Concentration (if solution): n/a

VEHICLE
- Amount(s) applied (volume or weight with unit): n/a
- Concentration (if solution): n/a
- Lot/batch no. (if required): n/a
- Purity: n/a

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 16 µL
- Concentration (if solution): n/a

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 16 µL
- Concentration (if solution): 5% aqueous solution of sodium dodecyl sulfate

Duration of treatment / exposure:

42 min

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean relative viability in %

Value:

106.04

Vehicle controls validity:

not applicable

Remarks:

The test item was applied neat to the tissues

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: none
- Direct-MTT reduction: none
- Colour interference with MTT: none

ACCEPTANCE OF RESULTS:

-Acceptability of the Quality Control Data of the Skin Model with Reference to Historical Batch Data:
The negative control OD values were 2.090, 2.057 and 2.044 and, thus, in the range of ≥0.8 and ≤3.0

-Acceptability of the Positive and Negative Control:
After treatment with the negative control (DPBS-buffer) the mean OD was 2.063 (standard deviation: 1.14%) and, thus, higher than the historically established boundary of 1.387.
After treatment with the positive control (5% aqueous solution of sodium dodecyl sulfate) the mean viability value was 1.20% (standard deviation: 7.43%) and, thus, lower than the historically established boundary of 3.58%.
The standard deviation of the negative control and the positive control was ≤18%, respectively.


-Test Substance Data Acceptance Criteria:
The standard deviation of the three tissues treated with the test item was 4.56% and, thus, ≤18%.
Therefore, the study fulfilled the validity criteria.

Group	Time / [min]	Mean OD	Mean Relative viability / [%]
Negative Control	42	2.515	100.00
Positive Control	42	0.021	0.83
Test Material	42	2.089	83.09

Interpretation of results:

GHS criteria not met

Remarks:

Non-corrosive

Conclusions:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 439. Under the conditions of the present study, the test item is not considered to be irritant to skin.

Executive summary:

The objective of the present study was to investigate the potential of the test item to induce skin corrosion in anin vitrohuman skin model.

This in vitro study was performed to assess the irritation potential of the test material by means of the Reconstructed Human Epidermis Test. The test consisted of a topical exposure of the test item to a human reconstructed skin model followed by a cell viability test. Cell viability was quantitatively measured by dehydrogenase conversion of MTT into a blue formazan salt after extraction from tissues. The percent reduction of cell viability in comparison to untreated negative controls was used to predict the skin irritation potential.

The tissue viability after treatment with the test item was 83.09% and, thus, higher than 50%, i.e. according to UN GHS classification the test item is considered as non-irritant to skin.

Under the conditions of the present study, the test item is not considered to possess a corrosive potential to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jul 28 - Oct 01, 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

2013

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Version / remarks:

Commission Regulation (EU) No. 1152/2010 amending, for the purpose of its adaption to technical progress, Regulation (EC) No. 440/2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006 of the European Parliament and the council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

PREPARATION OF THE TEST MATERIAL
The test item was prepared as a 20% (w/v) suspension in a 0.9% sodium chloride solution. The stability in the vehicle was not investigated. The test item preparation was administered within <1 hour after preparation.

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

TEST MATERIAL: 750 µL (i.e. 150mg/750µL)

NEGATIVE / VEHICLE CONTROL: 750 µL

Designation: 0.9% sodium chloride solution
Supplier: B. Braun Melsungen AG, Germany
Batch: 13445011
Released until: September 2016

POSITIVE CONTROL: 750 µL

Designation: Art. 814223
Synonym: Imidazole
Supplier: Merck KGaA, Germany
Batch: S6746923
Released until: August 31, 2018


Imidazole was dissolved with 0.9% sodium chloride solution to a concentration of 20% (w/v).

Duration of treatment / exposure:

240 minutes

Number of animals or in vitro replicates:

in vitro: triplicate design

Irritation parameter:

in vitro irritation score

Run / experiment:

Run 1 / Experiment 1

Value:

-0.2

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: No observations (e.g. tissue peeling, residual test chemical, non-uniform opacity patterns) were seen in a visually inspection of the corneas after treatment.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control:
After treatment with the negative control (0.9% sodium chloride solution) the calculated IVIS was 0.6 and, thus, within two standard deviations of the current historical mean of the negative control (IVIS: -1.3 – 3.3).

- Acceptance criteria met for positive control:
After treatment with the positive control (20% Imidazole) the calculated IVIS was 105.4 and, thus, also within two standard deviations of the current historical mean of the positive control (IVIS 74.2 – 138.9).

Therefore, the study fulfilled the acceptance criteria.

		Opacity	Permeability	IVIS
				per cornea	per group (mean value)	Standard deviation
Negative control	0.9% sodium chloride solution	0.566	0.007	0.656	0.6	0.1
		0.632	-0.001	0.617
		0.322	0.010	0.467
Positive control	Imidazole (20%)	79.434	1.370	99.977	105.4	6.0
		70.979	2.722	111.802
		63.046	2.759	104.435
Test item	Test material preparation	-0.880	0.016	-0.637	-0.2	0.4
		-0.503	0.038	0.065
		-0.128	0.017	0.120

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present study, the test item did not show an eye hazard potential. The test item is not requiring classification for eye irritation or serious eye damage (UN GHS: No Category).

Executive summary:

The objective of the present study was to examine the potential of the test item to induce serious eye damage in the BCOP assay. The BCOP assay with isolated fresh bovine corneas is an acceptedin vitromodel for ocular hazard assessment.

After a first opacity measurement of the untreated bovine corneas, 750 µL of the suspended test item, positive or negative control were applied on the corneas and incubated for 240 minutes. After the incubation phase the test item, the positive, and the negative control were rinsed from the corneas and the opacity was measured again.

After the opacity measurements, the permeability of the corneas was determined by application of a fluorescein solution for 90 minutes. The amount of fluorescein solution that crossed the cornea was measured spectrophotometrically.

The opacity and permeability assessments were combined to determine anIn VitroIrritancy Score (IVIS).

The IVIS obtained after treatment was -0.2 and, thus lower than 3,i.e.according to OECD 437 the test item is not requiring classification for eye irritation or serious eye damage (UN GHS: No Category).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro skin irritation (OECD 439)

The tissue viability after treatment with the test item was 83.09% and, thus, higher than 50%, i.e. according to UN GHS classification the test item is considered as non-irritant to skin.

In vitro eye irritation (OECD 437)

The IVIS obtained after treatment was -0.2 and, thus lower than 3,i.e.according to OECD 437 the test item is not requiring classification for eye irritation or serious eye damage (UN GHS: No Category).

Justification for classification or non-classification

The information provided shows that the test material does not need to classified for skin or eye irritation according to the EU Regulation (EC) No 1272/2008 on Classification,Labelling andPackaging of Substances and Mixtures.