2-{[(2-{3-[(4-{Ethyl[3-(vinylsulfonyl)phenyl]amino}-6-fluoro-heteromonocycle-2-yl)amino]-2- (hydroxy-O)phenyl}hydrazono-N2)(phenyl)methyl]diazenyl-N1}benzoato(6-)-O]cuprate(4-) sodium salt, polysulfo

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-12-14 to 2012-02-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test material

Test material information

Constituent 1

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Housing and Feeding Conditions:
- Semi barrier in an air-conditioned room
- Temperature: 18 +/- 3 °C (recommendations of TVT, GV-SOLAS)
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access to autoclaved hay and to Altromin 2123 maintenance diet for rabbits (lot no. XX), rich in crude fibre
- Free access to tap water (drinking water, municipal residue control, microbiological controls at regular intervals)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Housed in ABS - plastic rabbit cages, floor 4200 cm2
- Adequate acclimatisation period (at least 5 days) under laboratory conditions

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: water for injection

Controls:

no

Amount / concentration applied:

0.5 g per application site

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

Clinical Observation:
The animals were examined for signs of erythema and oedema 1 hour after the patch removal. For the determination of classification-relevant values, the animals were examined for signs of erythema and oedema 24, 48 and 72 hours after the patch removal. Dermal irritation was scored and recorded according to the grades in the table below. Any other signs such as hyperplasia, scaling, discolouration, fissures and scabs or any systemic effects were also recorded.
For the initial test in one animal, the test site was also examined immediately after the patch had been removed.

Evaluation of Results:
Individual reactions of each animal were recorded at each time of observation.
Nature, severity and duration of all lesions observed were described.
The body weight development was recorded at the start and at the end of the study.
On the basis of the test results, the test substance may be classified in one of the following classes in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC:
 Corrosive and assigned the symbol „C“ and the indication of danger corrosive in accordance with the following criteria:
- a substance or a preparation is considered to be corrosive if, when it is applied to healthy intact animal skin, it produces full thickness destruction of skin tissue on at least one animal during the test for skin irritation cited in Annex B.4 to Commission Regulation (EC) No. 440/2008, L 142 or during an equivalent method,
- classification can be based on the results of a validated in vitro test, such as that cited in Annex B.40 to Commission Regulation (EC) No. 440/2008, L 142 (skin corrosion: rat skin transcutaneous electrical resistance assay and human skin model assay),
- a substance or a preparation should also be considered corrosive if the result can be predicted, for example from strongly acid or alkaline reactions indicated by a pH of 2 or less or 11.5 or greater. However, where extreme pH is the basis for classification, acid/alkali reserve may also be taken into consideration. If consideration of alkali/acid reserve suggests the substance or preparation may not be corrosive then further testing should be carried out to confirm this, preferably by use of an appropriate validated in vitro test. Consideration of acid/alkali reserve should not be used alone to exonerate substances or preparations from classification as corrosive.
Risk phrases shall be assigned in accordance with the following criteria:
R35 Causes severe burns
- if, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to three minutes exposure, or if this result can be predicted.
R34 Causes burns
- if, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to four hours exposure, or if this result can be predicted,
- organic hydroperoxides, except where evidence to the contrary is available.
Notes:
Where classification is based on results of a validated in vitro test R35 or R34 should be applied according to the capacity of the test method to discriminate between these.
Where classification is based upon consideration of extreme pH alone, R35 should be applied.
 Irritant and assigned the symbol „Xi“ and the indication of danger „irritant“ in accordance with the criteria given below.
The following risk phrase shall be assigned in accordance with the criteria given:
R38 Irritating to skin
- Substances and preparations which cause significant inflammation of the skin which persists for at least 24 hours after an exposure period of up to four hours determined on the rabbit according to the cutaneous irritation test method cited in Annex B.4 to Commission Regulation (EC) No. 440/2008, L 142 (skin corrosion).
Inflammation of the skin is significant if:
(a) the mean value of the scores for either erythema and eschar formation or oedema formation, calculated over all the animals tested, is 2 or more; or
(b) in the case where the Annex B.4 (skin corrosion) test has been completed using three animals, either erythema and eschar formation or oedema formation equivalent to a mean value of 2 or more calculated for each animal separately has been observed in two or more animals.
In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating respective mean values.
Inflammation of the skin is also significant if it persists in at least two animals at the end of the time of observation. Particular effects e.g. hyperplasia, scaling, discoloration, fissures, scabs and alopecia should be taken into account. Relevant data may also be available from non-acute animal. These are considered significant if the effects seen are comparable to those described above.
- Substances and preparations which cause significant inflammation of the skin, based on practical observations in humans on immediate, prolonged or repeated contact.
- Organic peroxides, except where evidence to the contrary is available.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in Annex I of Regulation (EC) 1272/2008:
Skin corrosion, category 1 A, B, C:
Destruction of skin tissue, with subcategorisation based on exposure of up to 3 minutes (A), 1 hour (B), or 4hours (C). DANGER, corrosion symbol in diamond. Causes severe skin burns and eye damage.
Skin irritation, category 2:
Mean value of >=2.3 <= 4.0 for erythema / eschar or edema in at least 2 of 3 tested animals from gradings at 24, 48, and 72hours (or on 3 consecutive days after onset if reactions are delayed); inflammation that persists to the end of the (normally 14-day) observation period in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; in some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above. WARNING, exclamation mark in diamond. Causes skin irritation.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in GHS - Globally Harmonized System of Classification and Labelling of Chemicals, third revised edition, July 2009:
Skin corrosion, category 1 A, B, C: Destruction of skin tissue, with subcategorisation based on exposure of up to 3 minutes (A), 1 hour (B), or 4 hours (C). DANGER, corrosion symbol in diamond. Causes severe skin burns and eye damage.
Skin irritation, category 2: Mean value of >= 2.3 > 4.0 for erythema / eschar or oedema in at least 2 of 3 tested animals from gradings at 24, 48, and 72hours (or on 3 consecutive days after onset if reactions are delayed); inflammation that persists to end of the (normally 14-day) observation period. WARNING, exclamation mark in diamond. Causes skin irritation.
Skin irritation, category 3: Mean value of >= 1.5 < 2.3 for erythema / eschar or oedema in at least 2 of 3 tested animals from gradings at 24, 48, and 72 hours (or on 3 consecutive days after onset if reactions are delayed). WARNING, no symbol. Causes mild skin irritation.
No classification or labelling unless category 1 – 3 criteria are met.

Results and discussion

In vivo

Irritant / corrosive response data:

No irritant or corrosive effects were observed on the intact skin of the three female rabbits (strain NZW) after a contact time of 4 hours.

Other effects:

Neither mortalities nor significant clinical signs of toxicity were observed.
There were no significant body weight changes during the contact and observation period.

Any other information on results incl. tables

Table: Dermal Irritation Evaluation

Animal No.	Application Site	Irritation (hours after patch removal)
		1 hour		24 hours		48 hours		72 hours
		T	C	T	C	T	C	T	C
1	Erythema	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0
2	Erythema	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0
3	Erythema	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0

T = test item;       C = control

Table: Individual Data

Individual Systemic and Local Findings - Animal No. 1
Time after Patch Removal	Systemic Findings	Specific Local Findings (apart from erythema/oedema)	Comments
1 hour	nsf.	nsf	-
24 hours	nsf	nsf	-
48 hours	nsf	nsf	-
72 hours	nsf	nsf	-
Individual Systemic and Local Findings - Animal No. 2
Time after Patch Removal	Systemic Findings	Specific Local Findings (apart from erythema/oedema)	Comments
1 hour	nsf	nsf	-
24 hours	nsf	nsf	-
48 hours	nsf	nsf	-
72 hours	nsf	nsf	-

 

Individual Systemic and Local Findings - Animal No. 3
Time after Patch Removal	Systemic Findings	Specific Local Findings (apart from erythema/oedema)	Comments
1 hour	nsf	nsf	-
24 hours	nsf	blue coloured skin due to test item residues	-
48 hours	nsf	nsf	-
72 hours	nsf	nsf	-

nsf = no specific findings

Table: Absolute Body Weights in kg

	Animal No. 1	Animal No. 2	Animal No. 3
Start of Study (weight in kg)	3.6	3.2	3.4
End of Study (weight in kg)	3.8	3.3	3.4

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Under the conditions of the present study, the single dermal application of the test item FAT 40853/A TE to three rabbits at a dose of 0.5 g showed neither irritant nor corrosive effects.
Neither mortalities nor significant clinical signs of toxicity were observed.
In conformity with the EC criteria for classification and labelling requirements for dangerous substances and preparations according to Annex VI of Commission Directive 2001/59/EC , the test item FAT 40853/A TE does not have to be classified and has no obligatory labelling requirement for skin irritation.
According to Annex I of Regulation (EC) 1272/2008, the test item FAT 40853/A TE does not have to be classified and has no obligatory labelling requirement for skin irritation.
According to GHS (Globally Harmonized Classification System), the test item FAT 40853/A TE has no obligatory labelling requirement for skin irritation.

Executive summary:

Species/strain:                      New Zealand White Rabbits Crl: KBL (NZW)

Number of animals:              3

Duration of exposure:           4 hours

Amount of substance:           0.5 g per test site

Type of dressing:                  occlusive  [    ]  semi-occlusive  [ X ]

Vehicle (moistening):            aqua ad injectionem

First time of effects:              no effects observed

Last time of effects:              no effects observed

Reversibility of the
observed effects:                  no effects observed

Method: OECD 404
EC 440/2008, Method B.4
OPPTS 870.2500 1.1.             Conclusion

Under the conditions of the present study, the single dermal application of the test item FAT 40853/A TE to three rabbits at a dose of 0.5 g showed neither irritant nor corrosive effects.

Neither mortalities nor significant clinical signs of toxicity were observed.

In conformity with the EC criteria for classification and labelling requirements for dangerous substances and preparations according toAnnex VI of Commission Directive 2001/59/EC, the test item FAT 40853/A TE does not have to be classified and has no obligatory labelling requirement for skin irritation.

According to Annex I of Regulation (EC) 1272/2008, the test item FAT 40853/A TE does not have to be classified and has no obligatory labelling requirement for skin irritation.

According to GHS (Globally Harmonized Classification System), the test item FAT 40853/A TE has no obligatory labelling requirement for skin irritation.