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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-02-13 to 2012-03-29
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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals 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. 1610), 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:
water
Remarks:
Aqua ad injectionem, Diprom, lot no. 109521-1, expiry date 09/2013
Controls:
no
Amount / concentration applied:
A dose of 0.5 g of the test item was applied to each test site.
Duration of treatment / exposure:
The test item was held in contact with the skin throughout a 4-hour period.
At the end of the exposure period, the residual test item was removed with tap water.
Observation period:
All animals were observed for 72 hours after the patch removal.
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.

residue of the test item

-

24 hours

nsf

residue of the test item

-

48 hours

nsf

residue of the test item

-

72 hours

nsf

residue of the test item

-

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

residue of the test item

-

24 hours

nsf

residue of the test item

-

48 hours

nsf

residue of the test item

-

72 hours

nsf

residue of the test item

-

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

residue of the test item

-

24 hours

nsf

residue of the test item

-

48 hours

nsf

residue of the test item

-

72 hours

nsf

residue of the test item

-

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)

3.7

3.6

4.4

End of Study
(weight in kg)

3.7

3.7

4.5

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 40858/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 40858/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 40858/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 40858/A TE has no obligatory labelling requirement for skin irritation.
For details of the classification criteria, see Evaluation of Results.
Executive summary:

On the basis of the test results given below and in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC[6], the substance should be assigned the following risk phrase:

R38 irritating to skin

 

R34 causes burns

 

R35 causes severe burns

 

no phrase

X

On the basis of the test results given below and in conformity with the criteria given in Annex I of Regulation (EC) 1272/2008, the substance should be:

classified into category 1

 

classified into category 2

 

not classified

X

On the basis of the test results given below and in conformity with the criteria given in GHS (Globally Harmonized System of Classification and Labelling of Chemicals), the substance should be:

classified into category 1

 

classified into category 2

 

classified into category 3

 

not classified

X

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


 Average Scores – (24, 48, 72 hour reading)

Mean Value Irritation Scores

Animal No.

Mean 24 – 72 hours

Erythema

Oedema

1

0

0

2

0

0

3

0

0

Total Mean Value

0

0

Conclusion:

Under the conditions of the present study, the single dermal application of the test item FAT 40858/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 40858/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 40858/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 40858/A TE has no obligatory labelling requirement for skin irritation.

For details of the classification criteria, see Evaluation of Results.