Reaction mass of Benzeneethanol, 2-chloro-α-(1-chlorocyclopropyl)-α-(chloromethyl)- and Oxirane, 2-(1-chlorcyclopropyl)-2-[(2-chlorphenyl)methyl]-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 02 - April 26, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The guinea pig maximisation test was performed in accordance to the OECD guideline 406 and can be considered as a validated and reliable testing method for skin sensitization including a high statistical sample size and the challenge phase. In the beginning of 2001 the GPMT was the generally accepted in vivo model for the assessment of the skin sensitisation potential of substances in science and regulatory context. The adoption of the LLNA as a new Test Guideline is dated 24th April 2002.

Test material

Test material information

1

Reference substance name:

Reaction mass of Benzeneethanol, 2-chloro-.alpha.-(1-chlorocyclopropyl)-.alpha.-(chloromethyl)- and Oxirane, 2-(1-chlorcyclopropyl)-2-[(2-chlorphenyl)methyl]-

EC Number:

929-889-1

Cas Number:

not available

IUPAC Name:

Reaction mass of Benzeneethanol, 2-chloro-.alpha.-(1-chlorocyclopropyl)-.alpha.-(chloromethyl)- and Oxirane, 2-(1-chlorcyclopropyl)-2-[(2-chlorphenyl)methyl]-

Specific details on test material used for the study:

- Solubility and stability of the test substance in the solvent/vehicle: The stability of the test item in the vehicle was analytically verified for up to 2 hours.
- Treatment of test material prior to testing: The test item was formulated in polyethylene glycol 400. The formulations were visually described as solutions.

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Hsd Poc:DH

Sex:

female

Details on test animals and environmental conditions:

Animals:
-Reason for this choice: Species generally accepted by regulatory authorities for this type of study. The strain used has been shown to produce a satisfactory sensitization response using known sensitizers.
-Breeder: Harlan Winkelmann GmbH Laboratory, 33176 Borchen, Germany
-Number: 7 for the preliminary test, 30 animals for the main test.
-Age/weight: 5 - 6 weeks/284 -383 grams
-Acclimation: At least 5 d before the beginning of the study.
-Identification of the animals: Ear-tattoo.

Environmental conditions:
-Temperature: 22 ± 3°C
-Relative humidity: 40 to 60%
-Light/dark cycle: 12 h/ 12 h
-Ventilation: Approx >= 10 times per hour

Study design: in vivo (non-LLNA)

Induction

Route:

intradermal and epicutaneous

Vehicle:

polyethylene glycol

Concentration / amount:

Intradermal: 2.5% (= 10 mg test item/animal)
Topical: 100% (= 500 mg test item/animal)

Day(s)/duration:

once - the injection sites were visually assessed 2 and 7 days after the injections One week after the intradermal induction the topical (48-hour exposure period) was performed

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

No.:

#1

Route:

epicutaneous, semiocclusive

Vehicle:

polyethylene glycol

Concentration / amount:

25%

Day(s)/duration:

24 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

range finding group: 7 animals
test item group: 20 animals
control group: 10 animals

Challenge controls:

A patch loaded only with the vehicle was placed also on the right flank (cranial) as control.

Positive control substance(s):

yes

Remarks:

Vohr, H.-W.: Validation of the Magnusson - Kligman Maximization Test Method used by the Fachbereich Toxikologie, Bayer AG, performed in Guinea Pigs of the Strain Hsd:Poc:DH with Alpha-Hexylzimtaldehyd (BAYER-AG, PH 31458, November 2001).

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25 %

No. with + reactions:

16

Total no. in group:

20

Remarks on result:

positive indication of skin sensitisation

Results 2

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

25 %

No. with + reactions:

16

Total no. in group:

20

Remarks on result:

positive indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

no indication of skin sensitisation

Results 4

Reading:

2nd reading

Hours after challenge:

72

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

no indication of skin sensitisation

Results 5

Group:

positive control

Remarks on result:

positive indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Executive summary:

A study was conducted to assess the sensitizing potential of the test item in a guinea-pig maximisation test according to OECD Guideline 406 and EU method B.6.

The intradermal induction was performed using a 2.5% test item concentration in polyethylene glycol 400, and the topical induction was performed with a 100% test item concentration. After the intradermal induction the animals in the control group and in the test item group showed strong effects up to encrustation at the injection sites of the first induction. The challenge with the 25% test item formulation led to skin effects (grade 1 - 3) in 17 of 20 animals (85%) in the test item group and no skin effects were seen in the control group animals.