Azarbre

Endpoint summary

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:

3-9 June 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, to GLP

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Pre-incubated overnight (guideline recommends 18-24 h)

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: human derived epidermal keratinocytes

Strain:

not specified

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SkinEthic Laboratories, Lyon, France
- Age at study initiation: Not applicable
- Weight at study initiation: Not applicable
- Housing: Not applicable
- Diet (e.g. ad libitum): Not applicable
- Water (e.g. ad libitum): Not applicable
- Acclimation period: Not applicable

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not applicable
- Humidity (%): Not applicable
- Air changes (per hr): Not applicable
- Photoperiod (hrs dark / hrs light): Not applicable

IN-LIFE DATES: From: 3 June 2014 To: 9 June 2014

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 μL (26.3 μL/cm squared)
- Concentration (if solution): Not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable

Duration of treatment / exposure:

15 minutes

Observation period:

42 hours post-exposure incubation period

Number of animals:

Not applicable

Details on study design:

TEST SITE
- Area of exposure: 0.38 cm squared
- % coverage: Not applicable
- Type of wrap if used: Not applicable

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not applicable
- Time after start of exposure: Not applicable

SCORING SYSTEM: Not applicable

Irritation / corrosion parameter:

other: other: Relative viability (test item)

Value:

7.2

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 42.25 h. Max. score: 100.0. (migrated information)

Irritation / corrosion parameter:

other: other: Optical density (test item)

Value:

0.078

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 42.25 h. (migrated information)

Irritation / corrosion parameter:

other: other: Relative viability (positive control)

Value:

6.2

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 42.25 h. Max. score: 100.0. (migrated information)

Irritation / corrosion parameter:

other: other: Optical density (positive control)

Value:

0.068

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 42.25 h. (migrated information)

Irritation / corrosion parameter:

other: other: Relative viability (negative control)

Value:

100

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 42.25 h. Max. score: 100.0. (migrated information)

Irritation / corrosion parameter:

other: other: Optical density (negative control)

Value:

1.09

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 42.25 h. (migrated information)

Irritant / corrosive response data:

No data

Other effects:

No data

Item	OD of tissues	Mean OD	± SD of OD	Relative individual tissue viability	Relative mean viability/%	± SD of Relative mean viability/%
Negative control	1.138	1.090	0.049	104.4	100	4.5
	1.040			95.4
	1.093			100.3
Positive control	0.083	0.068	0.028	7.6	6.2	2.6
	0.035			3.2
	0.085			7.8
Test material	0.060	0.078	0.031	5.5	7.2	2.9
	0.060			5.5
	0.114			10.5

Mean optical density (OD) values, percentage viabilities and standard deviations (SD) for the negative control, positive control and test material. Mean viabilities are measured relative to the negative control (i.e. this is set at 100%).

The relative mean tissue viability for the positive control was 6.2% compared to the negative control, with a standard deviation of 2.6%. The mean optical density for the negative control was 1.090 and the standard deviation of the percentage viability was 4.5%. The standard deviation of the three test percentage viabilities was 2.9%. The acceptance criteria for negative, positive and test material were therefore satisfied.

Interpretation of results:

irritating

Remarks:

Migrated information Category 2 Criteria used for interpretation of results: EU

Conclusions:

In a guideline study (OECD TG 439), to GLP, the dermal irritation of azarbre was assessed using the in vitro human epidermis model. The test item was classified as irritant according to EU labelling regulations.

Executive summary:

In an OECD guideline study (TG 439), conducted according to GLP, the dermal irritation of azarbre was assessed using the in vitro human epidermis model. The quality acceptance criteria were satisfied for both the positive and negative controls and for the test matieral. The relative mean tissue viability of the test item was less than 50% of the negative control (7.2%) and was therefore classified as a category 2 irritant according to EU labelling regulations.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (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:

October 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to guideline. In compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: Chicken

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The eyes used in this assay were collected from c hickens obtained from a slaughterhouse (Etablissement Brun, 338 Etauliers, France) where they were killed for human consumption. Chicken age was generally 7 weeks old.
The eyes of the chickens were enucleated at Phycher.

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

30uL of test item was applied

Duration of treatment / exposure:

The test item was applied for 10 seconds so that it covered the entire surface of the cornea evenly. It was then rinsed from the eye with 20mL of physiological saline at ambient templerature.

Observation period (in vivo):

Treated corneas were evaluated pretreatment and after treatment 30, 75, 120, 180 and 240 minutes (+/- 5 minutes).

Number of animals or in vitro replicates:

3 enucleated chicken eyes

Details on study design:

Negative controls (physiological saline) and positive controls (5% Benzalkonium chloride) were included in this experiment.

Irritation parameter:

maximum mean total score (MMTS)

Remarks:

Corneal Opacity

Basis:

mean

Score:

ca. 1.3

Remarks on result:

other: Corresponds to ICE class II

Irritation parameter:

other: Mean score of fluorescein retention

Basis:

mean

Score:

ca. 0.7

Remarks on result:

other: Corresponds to ICE class II

Irritation parameter:

cornea opacity score

Remarks:

Maximal mean corneal swelling

Basis:

mean

Time point:

other: 240 minutes post-dose

Score:

ca. 12

Remarks on result:

other: Corresponds to ICE class II

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Executive summary:

The results obtained under the experimental conditions described, lead to the conclusion that Azarbre is not a corrosive / severe irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Justification for selection of skin irritation / corrosion endpoint:
Reliable in vitro K1 study

Justification for selection of eye irritation endpoint:
Reliable in vitro K1 study

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

Skin irritation not observed and therefore no classification required.

Eye irrtation was observed and the following classification apply:

EU DSD (67 -548 -EEC) Irritant requires symbol "Xi" risk phrase R38 "Irritating to Skin"

EU CLP and UN GHS Hazard statement H315 "Causes Skin Irritation" Category 2