Registration Dossier

Administrative data

Description of key information

Waiving of the eye irritation endpoint and rational for the classification as an eye irritation and as a respiratory irritant
Classification for the respiratory irritation endpoint

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2012-06-12 to 2012-06-22
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability is rated 1 because the study followed the standard guideline of reference OECD 404, which describes a procedure designed to evaluate this endpoint. The results were reviewed for reliability and assessed as valid, and the study was conducted under GLP condition.
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Elevage de Gérome (quartier Labaste - F40260 Linxe)
- Age at study initiation: 11 weeks old
- Weight at study initiation: between 2.20 kg and 2.46 kg
- Housing: Individually housed
- Diet (e.g. ad libitum): Ad libitum - Tap-water from public distribution
- Water (e.g. ad libitum): Ad libitum - Foodstuff (SDS-C15)
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 °C +/- 3°C
- Humidity (%): 30 to 70%
- Air changes (per hr): approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: distilled water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g of the test item as supplied, moistened beforehand with distilled water


VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 mL
Duration of treatment / exposure:
Exposure of 4 hours
Observation period:
Observation at 1, 24, 48 and 72 hrs after removal of the patch
Number of animals:
1 rabbit + 2 other rabbits.
Details on study design:
TEST SITE
- Area of exposure: 6 cm² per patch. an untreated area on the opposite flank served as control.
- Type of wrap if used: a strip of surfical adhesive tape under semi-occlusive dressing.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): rinsing with distilled water and liquid paraffin
- Time after start of exposure: at the end of the exposure period, i.e after 4h.

SCORING SYSTEM: Grading of Skin Reactions as displayed in the OECD No. 404, dated April 24th, 2002.
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Other effects:
A pink coloration, not preventing local observations, was noted between days 0 and 3 in all animals.

Individual skin reactions and average scores after 4 -hour exposure

OBSERVATIONS

INDIVIDUAL DATA

Total of individual data

Skin reaction

Observation time

Animal n°

Weight (kg)

Animal n°

Weight (kg)

Animal n°

Weight (kg)

A2132

(application date : 2012-06-12)

Start: 2.33

A2174

(application date : 2012-06-19)

Start: 2.34

A2177

(application date : 2012-06-19)

Start: 2.20

End: 2.46

End: 2.42

End: 2.32


Erythema and eschar

1 hour (D0)

0

0

0

0

24 hours

0

0

0

0

48 hours

0

0

0

0

72 hours*

0

0

0

0

Mean(24 to 72 hrs)

0.0

0.0

0.0

/

Oedema formation

1 hour (D0)

0

0

0

0

24 hours

0

0

0

0

48 hours

0

0

0

0

72 hours*

0

0

0

0

Mean(24 to 72 hrs)

0.0

0.0

0.0

/

Other reactions

1 hour (D0)

0

24 hours

0

48 hours

0

72 hours*

0

Mean(24 to 72 hrs)

0.0

0.0

0.0

/

* As no persistent reaction was observed 72 hours after the treatment, the study was ended.

²: pink coloration, not preventing local observations.

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The results obtained, in these experimental conditions, enable to conclude that the test item SEPISOL FAST RED SN must not be classified, according to the criteria for classification, packging and labelling of dangerous substances and preparations in compliance with the E.E.C. directives 67/548, 2001/59 and 99/45. No symbol or risk phrase is required.

In accordance with the regulation (EC) No. 1272/2008, the test item must not be classified. No signal word or hazard statement is required.
Executive summary:

The test item SEPISOL FAST RED SN was applied, as supplied, at the dose of 0.5 g, under semi-occlusive dressing during 4 hours on an undamaged skin area of three New Zealand rabbits. The experimental protocol was established from the OECD guideline No. 404 dated April 24th, 2002 and the test method B.4 of the Council regulation No. 440/2008 of 30 May 2008.

No cutaneaous reactions (erythema and oedema) were observed whatever the examination time (1, 24, 48 and 72 hours). A pink coloration, not preventing erythema assessment, was noted between days 0 and 3 in all animals.

The results obtained, in these experimental conditions, enable to conclude that the test item SEPISOL FAST RED SN must not be classified, according to the criteria for classification, packaging and labelling of dangerous substances and preparations in compliance with the E.E.C. directives 67/548, 2001/59 and 99/45. No symbol or risk phrase is required.

In accordance with the Regulation (EC) No. 1272/2008, the test item must not be classified. No signal word or hazard statement is required.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

The BCOP assay (realized on the Sepisol Fast Violet 3B –Eye irritation.001) apparently underestimates the irritancy potential. In fact the in vivo assay performed on a very similar dye, namely the Sepisol Fast Violet 2B (Eye irritation.002) shows a high irritancy level whereas the BCOP assay shows a moderate irritation.

Furthermore, because of lack of practice and perspective on this category of molecules, no comparison with an appropriate database could be performed. So the accuracy of BCOP assay and its applicability domain for this category of substance cannot be assessed and the results remain uncertain.

Besides the uncertainty of the applicability domain of the test, the results of the BCOP assay was also biased by the coloration of the cornea, thus it was not possible to distinguish the corneal opacity due to the coloration by the dye and the one due to real toxicity. This finding support the previous statement that the BCOP assay underestimates the results since a part of the irritation revealed by the corneal opacity measurement was partly due to the coloration.

Those identified limitations for the BCOP method (i.e. coloration of test material and uncertainty of the applicability) can be extrapolated to the other in vitro tests method (validated or not), since (1) their current validation database did not allow an adequate evaluation of organic dyes class and (2) the coloring property of the dye is most likely to interfere with the implementation of the test protocols and/or the interpretation of the results. Therefore, the realization of an in vitro assay, to test the eye irritancy potency of the Sepisol Fast Red SN is not technically feasible.

The Sepisol Fast Red SN is a result of a reaction of co-precipitation between two ionic units, as well as all the other organic dyes cited in the studies Eye irritation 001 – 002 and 003.

The cationic part is represented by the so-called Basic Red 1, commonly known as the Rhodamine 6G (CAS n°989-38-8 / EC n°213-584-9) and the anionic part by the so-called Dowfax 2A1 (CAS n°28519-02-0/EC n°249-063-8 and CAS n°25167-32-5/EC n°246-688-8).

Based on a bibliographic searches, both of the ionic units are considered injurious for the eyes (Eye irritation 004 to 007).

The Sepisol Fast Violet 881239 is also constituted of the Dowfax 2A1 as the anionic part and of a cationic part named Basic Violet 1. As stated in the handbook and the publication cited in the Eye irritation 004 and 007 respectively, cationic dyes are known to be injurious to the eye. Thus the Basic Violet 1 is considered irritant for the eye, as the Rhodamine 6G.

And in the in vivo eye irritation test performed for the Violet 881239 (Eye irritation 003) the dye was found to be irreversibly highly irritating.

 

Based on all those data, it is reasonably expected that the Sepisol Fast Red SN will also be highly irritating to the eyes of rabbits and it should be directly classified “Eye damage category 1” in order to avoid an unnecessary and painful in vivo test on rabbit eyes.


Effects on eye irritation: highly irritating

Effects on respiratory irritation: irritating

Justification for classification or non-classification

Irritation of the respiratory tract:

The Sepisol Fast Red SN is a powder exhibiting a fine granulometry. The particles size distribution has been determined by laser diffraction. 90 % of the particles size stands below 232.35 µm, 50% below 59.69 µm and 10% below 6.065 µm. MMAD is epxected to be less than 100 µm.

The size of 81.2% of the total sample is less than 125 µm and most probably the MMAD is below 100 µm.

It is then expected that this powder will be present all along the respiratory tract and will penetrate deeply in the lungs. The Sepisol Fast Red SN as been classified as eye damage category 1. The effect did not seem to be caused by a mechanical irritation. As a result of the previous reasons Sepisol Fast Red SN is foreseen to be a possible irritant for the mucous membrane lining the respiratory tract. Therefore it has been classified accordingly: STOT category 3 - Inhalation; H335.

For the eyes irritating property:

To avoid an unnecessary in vivo test on rabbits’eyes, the Sepisol Fast Red SN has been directly classified “Eye damage 1 – H318” based on a weight of evidence approach discussed above.