Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
other information
Study period:
November 11 to 19, 1985
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Well documented and reported study, conducted equivalent or similar to internationally accepted technical guideline in recognized industrial research organization. A quality assurance inspection report with reference to GLP, but not a GLP compliance statement, was included in the study report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report date:
1986

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
of 1984
Deviations:
yes
Remarks:
exposure period was 24 hours instead of 4 hours
GLP compliance:
no
Remarks:
but Quality Assurance statement with reference to IKS GLP guidance document was included in the report

Test material

Constituent 1
Chemical structure
Reference substance name:
2,6-dichloro-N-phenylaniline
EC Number:
239-349-0
EC Name:
2,6-dichloro-N-phenylaniline
Cas Number:
15307-93-4
Molecular formula:
C12H9Cl2N
IUPAC Name:
2,6-dichloro-N-phenylaniline
Details on test material:
- Name of test material (as cited in study report): PBS 3247.2 (Voltamin)
- Description: yellowish to brown needles
- Expiration date of the lot/batch: December 1985
- Storage conditions: room temperature
Analyzed by Pharmaceuticals Production: Test Article Data Sheet of September 30, 1985


Test animals

Species:
rabbit
Strain:
other: albino, Chbb:HM (SPF)
Details on test animals or test system and environmental conditions:
- Animal supplier: Thomae, FRG
- Age: Approximately 4 - 6 months.
- Number and Sex: 3 males and 3 females
- Weight on day of treatment (before application): Minimum 1.69 kg, maximum 2.20 kg.
- Housing: Individual housing in metal cages.
- Diet (ad libitum): Commercially available pelleted standard diet (NAFAG No. 814, batch 73/85, analysed by the
manufacturer, NAFAG, Gossau, Switzerland).
- Water (ad libitum): Tap water. (Periodically analysed for compliance with Swiss drinking water specifications) - Acclimation period: Approximately 2 weeks under laboratory conditions.


ENVIRONMENTAL CONDITIONS

Air conditioned room:
- Temperature (°C): 18 ± 2°C
- Relative Humidity (%): 60 ± 10%
- Photoperiod: 14 hrs light/day


Test system

Type of coverage:
occlusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
0.5 g test material per approx. 6 cm2 skin per animal.
The unchanged test material was ground to a fine powder using a mortar. The ground test material was loaded on gauze patches moistened with distilled water.
Duration of treatment / exposure:
24 hours
Observation period:
From prior to treatment (i.e. pretest) until 7 days post patch removal
Number of animals:
6 (3 males and 3 females)
Details on study design:
TEST SITE PREPARATION:

3 days before treatment, both flanks of the rabbits were shaved with an electric clipper and an electric shaver.

- Area of treated skin: Approx. 6 cm2 (= 2.5 x 2.5 cm gauze patch).
- Type of wrap used: Gauze patches were covered with impermeable foil and fastened to the rabbit body with elastic bandage and adhesive tape.

TEST MATERIAL AND DOSE PREPARATION

- Administration Volume/animal (i.e. one gauze patch of 2.5 x 2.5 cm per animal):
Test material was ground to a fine powder using a mortar. 0.5 g of ground test material was loaded on a gauze patch moistened with distilled water.

TREATMENT PERIOD AND REMOVAL OF TEST SUBSTANCE

Epicutaneous, occlusive treatment lasted 24 hours. Then the dressings were removed and residual test material was gently washed off the skin with warm water

SCORING SYSTEM:

The treated skin patches were evaluated at pretest and 30 minutes 24, 48 and 72 hours and 7 days post patch removal. Skin reactions were evaluated for erythema/eschar formation and edema according to Draize (see Table 1). In addition, scaling and any other abnormalities were recorded.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Remarks:
according to Draize as defined above
Basis:
mean
Remarks:
over 6 animals and 3 time points
Time point:
other: 24, 48, 72 h
Score:
2.17
Max. score:
8
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Full reversibility of erythema/ edema in 4 of 6 animals. Marked improvement in the other two animals. 5 of 6 animals showed skin skaling or slight skin skaling on the treated flank on day 7 (i.e. at 7 days post patch removal).
Irritation parameter:
erythema score
Remarks:
according to Draize as defined above
Basis:
mean
Remarks:
over 6 animals and 3 time points
Time point:
other: 24, 48, 72 h
Score:
1.56
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Full reversibility of erythema in 4 of 6 animals. Improvement in the other two animals. 5 of 6 animals showed skin skaling or slight skin skaling on the treated flank on day 7.
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
mean
Remarks:
over 6 animals and 3 time points
Time point:
other: 24, 48, 72 h
Score:
0.61
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: 5 of 6 animals showed skin skaling or slight skin skaling on the treated flank on day 7.
Irritation parameter:
erythema score
Remarks:
according to Draize as defined above
Basis:
animal: male no. 28
Time point:
other: 24, 48, 72 h
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Skin scaling on treated flank at 48 h, 72 h and 7 days post patch removal
Irritation parameter:
erythema score
Remarks:
according to Draize as defined above
Basis:
animal: male no. 29
Time point:
other: 24, 48, 72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Skin skaling on treated flank on day 7
Irritation parameter:
erythema score
Remarks:
according to Draize as defined above
Basis:
animal: male no. 30
Time point:
other: 24, 48, 72 h
Score:
2.67
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Skin scaling on treated flank at 72 h and 7 days post patch removal.
Irritation parameter:
erythema score
Remarks:
acccording to Draize as defined above
Basis:
animal: female no. 58
Time point:
other: 24, 48, 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Skin scaling, slight in degree, on treated flank on day 7
Irritation parameter:
erythema score
Remarks:
according to Draize as defined above
Basis:
animal: female no. 59
Time point:
other: 24, 48, 72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritation parameter:
erythema score
Remarks:
according to Draize as defined above
Basis:
animal: female no. 60
Time point:
other: 24, 48, 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Skin skaling on treated flank on day 7
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
animal: male no. 28
Time point:
other: 24,48,72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other: Skin scaling on treated flank at 48 h, 72 h and 7 days post patch removal
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
animal: male no. 29
Time point:
other: 24,48,72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Skin skaling on treated flank on day 7
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
animal: male no. 30
Time point:
other: 24,48,72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Skin scaling on treated flank at 72 h and 7 days post patch removal
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
animal: female no. 58
Time point:
other: 24,48,72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
other: Skin scaling, slight in degree, on treated flank on day 7
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
animal: female no. 59
Time point:
other: 24,48,72 h
Score:
0
Max. score:
4
Reversibility:
other: Female no. 59 was free from edema throughout the study
Irritation parameter:
edema score
Remarks:
according to Draize as defined above
Basis:
animal: female no. 60
Time point:
other: 24,48,72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other: Skin skaling on treated flank on day 7
Irritant / corrosive response data:
Erythema and edema result tables for each individual animal are presented under "Remarks on results including tables and figures"
Other effects:
Yellow-brown discoloration of small areas of the treated flank was seen in one animal (female No. 60) at 24, 48 and 72 h after patch removal having fully reversed by 7 days.

Skin scaling or slight skin scaling, was seen in 5 of 6 animals (male Nos. 28, 29, 30 and female nos. 58 and 60) on the treated flank at 48 h, 72 h and/or 7 days after patch removal having not reversed by 7 days in any of these animals.

Any other information on results incl. tables

 

Table 2: Gradings of Skin Reactions Attained for each Male Animal at Different Observation Time Points

Pretest /
Time after Patch Removal

Pretest

0.5 h

24 h

48 h

72 h

7 days

Animal (Identification) No.

28 29 30

28 29 30

28 29 30

28 29 30

28 29 30

28 29 30

Erythema / Eschar

0   0   0

1   1   2

2   2   3

1   2   3

1   2   2

0   1   1

Edema

0   0   0

1   1   1

1   1   1

1   1   1

0   1   1

0   0   0

 

 

 

Table 3: Gradings of Skin Reactions Attained for each Female Animal at Different Observation Time Points

Pretest /
Time after Patch Removal

Pretest

0.5 h

24 h

48 h

72 h

7 days

Animal (Identification) No.

58 59 60

58 59 60

58 59 60

58 59 60

58 59 60

58 59 60

Erythema / Eschar

0   0   0

2   1   3

1   1   3

1   0   2

1   0   1

0   0   0

Edema

0   0   0

1   0   1

1   0   1

0   0   1

0   0   0

0   0   0

 

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the attained primary skin irritation index of 2.4 reflecting “moderate irritation” and the fact that the affected skin had not fully reversed to normal skin by termination of the study, classification of the test material as “R38 Irritating to skin” according to EU DIRECTIVE 67/548/EEC and as “Category 2, irritant” according to REGULATION (EC) 1272/2008) seems to be appropriate.
Executive summary:

PBS 3247.2 (Voltamin) was tested for skin irritation similar to EU Method B.4 (Acute Toxicity: Skin Irritation) of 1984 (Dir. 84/449/EEC) in albino rabbits [Chbb:HM (SPF)]. The study was more comprehensive than foreseen in this test guideline in that six animals were exposed to the test material for 24 hours (instead of 4 hours).

 

Reliability grade 1 was assigned to the study. It was not conducted in compliance with GLP, but a Quality Assurance statement with reference to an IKS GLP guidance document*was included in the study report.

 

Intact shaved skin of each animal (3 males and 3 females) was treated on one flank by epicutaneous occlusive administration of 0.5 g of ground test material loaded on gauze patches (2.5 x 2.5 cm), the latter moistened with distilled water. The patches were covered with impermeable foil and fastened to the rabbit body with elastic bandage and adhesive tape. Shaved skin on the other flank served as a control. Skin reactions were recorded at pretest and 30 minutes 24, 48 and 72 hours and 7 days post patch removal and were evaluated for erythema/eschar formation and edema according to Draize. In addition, scale formation and local discoloration of the skin were recorded.

 

There was no mortality. General signs of toxicity or relevant changes in body weight were not evident. Erythema up to grade 3 (moderate to severe erythema) were seen in all of six animals. In addition, very slight edema (Grade 1, barely perceptible) and scale formation on the treated flank were recorded in five of them and yellow-brown discoloration of small areas of the treated flank in one of them. At termination of the study, 7 days after patch removal, edema and discoloration of the skin had fully disappeared in all affected animals, whilst erythema had fully reversed in four and partly reversed in the other two of six animals and scale formation was evident in five animals.

 

The primary skin irritation index (i.e. sum of mean irritation scores read 30 minutes and 48 hours after patch removal for erythema and edema divided by 2) attained during this study was 2.4 (maximum possible value: 8) reflecting “moderate irritation” according to the skin irritation/corrosion classification scheme adopted in the study report. However, the mean Draize scores (24, 48, 72 h over 6 animals) were 1.56 for erythema and 0.61 for edema. For three of six animals, the respective mean score for erythema (24, 48, 72 h) was ≥ 2, whilst for the other three animals the respective erythema and for all animals the respective edema score were < 2.

 

Based on the attained primary skin irritation index reflecting “moderate irritation” and the fact that the affected skin had not fully recovered by termination of the study, classification of the test material as “R38 Irritating to skin” according to EU DIRECTIVE 67/548/EEC and as “Category 2, irritant” according to REGULATION (EC) 1272/2008) seems to be appropriate.

 -----------------------------------------------------------------------------------------

*"Wegleitung der IKS betreffend gute Laboratoriumspraxis für nichtklinische Laborversuche, Unterabschnitt B.4.a.v."