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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

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:
21 May - 27 July 2002
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
/ the test group consisted of 5 instead of 10 animals; Titermax was used as adjuvants; grading scale differs from OECD 406, limited data on test article given (expiration date, purity, storage)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report date:
2002

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
17 July 1992
Deviations:
yes
Remarks:
/ the test group consisted of 5 instead of 10 animals; Titermax was used as adjuvants; grading scale differs from OECD 406, limited data on test article given (expiration date, purity, storage)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was conducted in 2002, where no validated protocol for the LLNA was available.

Test material

Constituent 1

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Remarks:
SPF Hartley guinea pig Aai: (HA) Outbred
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Microbiological status of animals: Animals were checked for respiratory or intestinal disease, skin eruptions, mucosal membrane irritation, postural difficulties and general condition.
- Age at study initiation: 4 - 6 weeks
- Weight at study initiation: 386 - 423 g (Test group) 368 - 419 g (Control group)
- Housing: animals were housed in stainless steel cages (no further information available)
- Diet: Lab Diet Certified Guinea Pig Diet (#5026), ad libitum
- Water: ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15.6 - 29.4
- Humidity (%): 30 - 70%
- Photoperiod (hrs dark / hrs light): 12/ 12
- IN-LIFE DATES: From: 21 May To: 27 July 2002

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100% (v/v)
0.1 mL/ injection site
positive control: 25%
Day(s)/duration:
single injection
Adequacy of induction:
other: The test substance did not exhibit skin irritating properties in the screening test. Thus, undiluted test substance was used for intradermal injection.
Route:
epicutaneous, semiocclusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100 % (v/v)
2 x 4 cm patch spread with 0.5 mL
Day(s)/duration:
48 h
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
Challenge
No.:
#1
Route:
epicutaneous, semiocclusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100 %
0.4 mL applied via 25 mm Hilltop Chamber
Day(s)/duration:
1
Adequacy of challenge:
other: non-irritant concentration
No. of animals per dose:
3 males and 2 females (main test)
2 males and 2 females (epicutaneous pretest)
1 male (intradermal pretest)
Details on study design:
RANGE FINDING TESTS:
- intradermal injections:
site 1: 5% test substance (50/50 emulsion of TiterMax (TM) and distilled water)
site 2: 100%
site 3: 50% test substance (in distilled water)
Injections (0.1 mL of each) were made between shaved shoulder blades of one guinea pig.

- epidermal applications: 10, 25, 50 and 100 % (in distilled water). 0.4 mL of the test article were applied to close-clipped trunks of each of four guinea pigs via a 25 mm Hilltop Chamber (with the cotton patch). An elastic tape (Elastoplast, Beiersdorf Inc., Norwalk, USA) was wrapped around the animal, which was lined to pure latex dental dam (HCM - Hygienic Corporation, Malaysia). After 24 h, dressings were removed. The remaining test substance was wiped away with ethanol 45 h after application. Skin reactions were evaluated 3 and 24 h after the removal of the test substance.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: 1:1 mixture (v/v) TM/ distilled water
Injection 2: 100 % test substance
Injection 3: 100 % test substance, emulsified in a 1:1 mixture (v/v) TM/ distilled water

Epicutaneous:
100% test substance

- Control group: was not co-treated during induction phase
- Site: shoulder area (intradermal + epicutaneous)
- Frequency of applications: every 7 days
- Duration: days 0 - 9
- Concentrations: 100% (intradermal + epicutaneous)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day of challenge: 21
- Exposure period: 24 h
- Test groups: test substance and positive control substance
- Control group: 100% test substance
- Site: shaved flank of respective animal
- Concentrations: 100%
- Evaluation (hr after challenge): 48, 72 and 96 h
Challenge controls:
the control group is actually a challenge control
Positive control substance(s):
yes
Remarks:
1 -Chloro-2,4-Dinitrobenzene, Lot# 100K1323

Results and discussion

Positive control results:
1-Chloro-2,4-Dinitrobenzene was used as positive control substance in the study. For intradermal induction, the following injetions were made:
site 1: TiterMax®/ distilled water (1:1)
site 2: 5% test substance (in corn oil)
site 3: 1% test substance (in TiterMax®/ distilled water emulsion (1:1))
On day 7, 25% 1-Chloro-2,4-Dinitrobenzene solved in petrolium jelly was applied for epidermal induction. For challenge, 0.05% 1-Chloro-2,4-Dinitrobenzene dissolved in pertoleum jelly was applied.

At the 48 h reading time point, 4/5 animals revealed erythema graded with 1 (3/4 animals, very slight erythema) or 2 (1/4 animals, well-defined erythema). 24 h later, erythema were still visible in all 4 animals scored with grade 1 (very slight erythema). Very slight erythema (scored with grade 1) and questionable erythema were observed in 1/5 and 3/5 animals at the 72 h reading time point, respectively. No edema were observed.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
intradermal induction: 0%; challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
intradermal induction: 100% challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Animals were treated with a 10 % Sodium-Lauryl-Sulphate solution 24 h prior to the epicutaneous induction.
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
Dose level:
intradermal induction: 5% challenge: 0.05%
No. with + reactions:
4
Total no. in group:
5
Remarks on result:
other: One additional animal showed a questionable erythema.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
intradermal induction: 0%; challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
test chemical
Dose level:
intradermal induction: 100% challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Animals were treated with a 10 % Sodium-Lauryl-Sulphate solution 24 h prior to the epicutaneous induction.
Reading:
2nd reading
Hours after challenge:
72
Group:
positive control
Dose level:
intradermal induction: 5%; challenge: 0.05%
No. with + reactions:
4
Total no. in group:
5
Reading:
other: 3rd reading
Hours after challenge:
96
Group:
negative control
Dose level:
intradermal induction: 0% challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
other: 3rd reading
Hours after challenge:
96
Group:
test chemical
Dose level:
intradermal induction: 100% challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Animals were treated with a 10 % Sodium-Lauryl-Sulphate solution 24 h prior to the epicutaneous induction.
Reading:
other: 3rd reading
Hours after challenge:
96
Group:
positive control
Dose level:
intradermal induction: 5% challenge: 0.05%
No. with + reactions:
1
Total no. in group:
5
Remarks on result:
other: 3 additional animals showed questionable erythema.

Any other information on results incl. tables

Pre-tests

Intradermal injection

1 male was injected with 100 µL of the test substance per test site as follows:

Site    Dosage

1         5% in a 50/50 emulsion of TiterMax® and distilled water

2         100% (undiluted)

3         50% in distilled water

 

Table 1: Intradermal injection

Animal No.

Sex

Test site

Concentration [%]

Score after 24 h

Score after 48 h

Erythema

Oedema

Erythema

Oedema

1

M

1

5

2

1

2

1

2

100

0

0

0

0

3

50

1

0

0

0

 

Epicutaneous application

4 animals (2 males/2 females) were treated with different concentrations (10, 25, 50, 100% in distilled water) of the test substance. At the 48 h reading time point, 2 males showed a questionable erythema at skin areas treated with 50 and 100% test substance. In one male, the questionable erythema was still visible 24 h later (72 h reading time points) at both concentrations. There were no effects (oedema or erythema) visible in the other animals or using the other 2 concentrations, neither at the 48 nor after 72 reading time point.

Table 2: Body weights summary

 

 

Initial weight [g]

Terminal weight [g]

Test group

M

400

519

388

466

423

519

F

386

464

404

497

Control group

M

386

524

393

514

386

466

F

419

529

368

514

Test group (Positive control)

M

442

518

402

541

450

580

F

384

504

392

498

Control group (Positive control)

M

420

556

386

554

360

498

F

430

562

436

560

 

The body weight gain of the the animals was not affected adversely during the study.

Applicant's summary and conclusion

Interpretation of results:
other: CLP/ EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
CLP: not classified