Registration Dossier

Diss Factsheets

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 01 - October 25, 1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report date:
1981

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
Batchno and purity were not given, and limited reported housing conditions.
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
study was already available before entry into force of REACH

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2'-azobis[2-methylpropionamidine] dihydrochloride
EC Number:
221-070-0
EC Name:
2,2'-azobis[2-methylpropionamidine] dihydrochloride
Cas Number:
2997-92-4
Molecular formula:
C8H18N6.2ClH
IUPAC Name:
2,2'-diazene-1,2-diylbis(2-methylpropanimidamide) dihydrochloride
Details on test material:
- Name of test material (as cited in study report): V-50

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Dean Dual, Luxemburg, Wisconsin
- Age at study initiation: young adults
- Weight at study initiation: 331-421 g
- Housing: Individually in stainless-steel cages
- Diet (e.g. ad libitum): Continuous access to Purina Guinea Pig Chow
- Water (e.g. ad libitum): Continuous access to water
- Acclimation period: At least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data, temperature-controlled rooms
- Humidity (%): no data, humidity-controlled rooms
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: From: 01 October 1981 To: 22 October 1981

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
petrolatum
Concentration / amount:
The Freund's Complete Adjuvant Solution used in all intradermal injections was prepared by adding 2.5 ml of sterile water for injection in approximately 1.0 ml increments to 2.5 ml of Freund's Adjuvant.

The 5% solution of test material in sterile water was prepared by taking 0.5g of V-50 and adding sterile water for injection to a total volume of 10.0 ml.

The 5% solution of test material in Freund's Adjuvant was prepared by adding 2.5 ml of a 10% w/v mixture of V-50 in sterile water to 2.5 ml of Freund's Adjuvant solution. The test material was added in approximately 1.0 ml increments.

For the topical induction and challenge procedure, the test material was administered as 50% w/w and 25% w/w mixtures, respectively, in petrolatum.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
The Freund's Complete Adjuvant Solution used in all intradermal injections was prepared by adding 2.5 ml of sterile water for injection in approximately 1.0 ml increments to 2.5 ml of Freund's Adjuvant.

The 5% solution of test material in sterile water was prepared by taking 0.5g of V-50 and adding sterile water for injection to a total volume of 10.0 ml.

The 5% solution of test material in Freund's Adjuvant was prepared by adding 2.5 ml of a 10% w/v mixture of V-50 in sterile water to 2.5 ml of Freund's Adjuvant solution. The test material was added in approximately 1.0 ml increments.

For the topical induction and challenge procedure, the test material was administered as 50% w/w and 25% w/w mixtures, respectively, in petrolatum.
No. of animals per dose:
Treated group 15 animals, and a naive control group, positive control group and vehicle group of four animals each.
Details on study design:
RANGE FINDING TESTS: A range-finding study to determine the irritation threshold of the test material was conducted. Based upon these results, the test material was administered as 50% w/w mixture in petrolatum for the topical induction and as 25% w/w in petrolatum for the challenge application.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: two
- Exposure period: intradermal injections: 7 days, topical induction: 48 hours
- Test groups: 50% w/w mixture of V-50 in petrolatum
- Control group: naive control group, positive control group (0.1% w/v mixture of 2,4-Dinitrochlorobenzene in sterile water) and vehicle control group (sterile 0.9% saline)
- Site: A 4.0x6.0 com area was clipped along the midline across the shoulder region,
- Frequency of applications: one

B. CHALLENGE EXPOSURE
- No. of exposures: one
- Day(s) of challenge: two weeks after topical induction the animals received a challenge dose for 24 hours.
- Exposure period: 24 hours
- Test groups: 25% w/w mixture of V-50 in petrolatum
- Control group: positive control group and vehicle control group.
- Site: The hair was removed from a 5.0x5.0 cm area on the flank by clipping and as reported at the induction exposure.
- Evaluation (hr after challenge): 24 and 48 hours following patch removal the test sites were examined for erythema and edema.

Positive control substance(s):
yes
Remarks:
The 0.1% solution of 2,4-Dinitrochlorobenzene in sterile water was prepared by adding 0.10 g of 2,4-Dinitrochlorobenzene to 100 ml of sterile water. The solution was mildly heated and thoroughly stirred prior to use.

Results and discussion

Positive control results:
A sensitization reaction to the positive control was exhibited by all animals following the challenge application. All animals responded with a moderate and diffuse redness at 24 hours. At the 48 hour observation a moderate and diffuse redness was seen in one animal and three animals had a mild redness.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
Challenge dose 25% w/w
No. with + reactions:
13
Total no. in group:
15
Clinical observations:
Ten animals reacting to the challenge application exhibited a moderate and diffuse redness. Three animals exhibited a mild redness. Two test animals were found dead on Day 11 of the study.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Challenge dose 25% w/w. No with. + reactions: 13.0. Total no. in groups: 15.0. Clinical observations: Ten animals reacting to the challenge application exhibited a moderate and diffuse redness. Three animals exhibited a mild redness. Two test animals were found dead on Day 11 of the study..
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
Challenge dose 25% w/w
No. with + reactions:
13
Total no. in group:
15
Clinical observations:
Six animals continued to exhibited a moderate and diffuse redness while the remaining seven animals exhibited a mild response.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Challenge dose 25% w/w. No with. + reactions: 13.0. Total no. in groups: 15.0. Clinical observations: Six animals continued to exhibited a moderate and diffuse redness while the remaining seven animals exhibited a mild response..
Reading:
1st reading
Hours after challenge:
24
Group:
other: untreated
Dose level:
Challenge dose 25% w/w
No. with + reactions:
3
Total no. in group:
4
Clinical observations:
Three animals responded to the challenge application with a mild redness.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: other: untreated. Dose level: Challenge dose 25% w/w. No with. + reactions: 3.0. Total no. in groups: 4.0. Clinical observations: Three animals responded to the challenge application with a mild redness..
Reading:
2nd reading
Hours after challenge:
48
Group:
other: untreated
Dose level:
Challenge dose 25% w/w
No. with + reactions:
1
Total no. in group:
4
Clinical observations:
A mild redness was observed in one animal.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: untreated. Dose level: Challenge dose 25% w/w. No with. + reactions: 1.0. Total no. in groups: 4.0. Clinical observations: A mild redness was observed in one animal..
Reading:
1st reading
Hours after challenge:
24
Group:
other: vehicle control
Dose level:
Sterile 0.9% Saline
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
None of the guinea pigsexhibited a reaction to the challenge dose.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: other: vehicle control. Dose level: Sterile 0.9% Saline. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: None of the guinea pigsexhibited a reaction to the challenge dose..
Reading:
2nd reading
Hours after challenge:
48
Group:
other: vehicle control
Dose level:
Sterile 0.9% Saline
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
None of the guinea pigs exhibited a reaction to the challenge dose.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: vehicle control. Dose level: Sterile 0.9% Saline. No with. + reactions: 0.0. Total no. in groups: 0.0. Clinical observations: None of the guinea pigsexhibited a reaction to the challenge dose..
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.1% w/v Dinitrochlorobenzene
No. with + reactions:
4
Total no. in group:
4
Clinical observations:
All animals responded with a moderate and diffuse redness.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.1% w/v Dinitrochlorobenzene. No with. + reactions: 4.0. Total no. in groups: 4.0. Clinical observations: All animals responded with a moderate and diffuse redness..
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.1% w/v Dinitrochlorobenzene
No. with + reactions:
4
Total no. in group:
4
Clinical observations:
A moderate and diffuse redness was seen in one animal and three animals had a mild redness.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.1% w/v Dinitrochlorobenzene. No with. + reactions: 4.0. Total no. in groups: 4.0. Clinical observations: A moderate and diffuse redness was seen in one animal and three animals had a mild redness..

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based upon the results obtained, the substance is considered a strong skin sensitizer in guinea pigs.
According to Guidance on information requirements and chemical safety assessment Chapter R.8, Appendix R.8-10, the substance is allocated to the moderate potency category.
Executive summary:

This study was designed to evaluate the skin sensitization properties of the substance using the Maximization test. The study consisted of a treated group of 15 animals, and a naive control group, positive control group (Dinitrochlorobenzene) and vehicle group (0.9% saline) of four animals each. In the test substance group, 0.05 ml test substance was injected intradermally on each side of the midline across the shoulder region of guinea pigs for the first induction. After 7 days, they were secondly induced with a 50% w/w mixture of V-50 in petrolatum for 48 hours. After two weeks, the right flank was challenged with 25% w/w test substance for 24 hours. Vehicle and positive control group were treated similarly. As a result of the study, all surviving animals of the test substance group and all animals of the positive control group exhibited a sensitization reaction following the challenge application. None of the animals exhibited a reaction in the vehicle control group. According to the results, it is concluded that the substance has strong skin sensitization properties in guinea pigs. Classification: Skin Sens. Cat. 1, May cause an allergic reaction (H317).