1-(3-chlorophenyl)-4-(3-chloropropyl)piperazinium chloride

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data is from peer reviewed journals

Data source

Reference

Reference Type:

publication

Title:

Evaluation of Skin Sensitization and Cross Reaction of nine Alkyleneamines in the Guinea pig maximization test

Author:

HON- WING LEUNG, CAROL S. AULETTA

Year:

1997

Bibliographic source:

Journal of Toxicology: Cutaneous and Ocular Toxicology 16, no. 3 (1997): 189-195

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Magnusson and Kligman

Principles of method if other than guideline:

The dermal sensation potential of the test chemical was evaluated in guinea pigs following the method described by Magnusson and Kligman

GLP compliance:

not specified

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

2-piperazin-1-ylethylamine

EC Number:

205-411-0

EC Name:

2-piperazin-1-ylethylamine

Cas Number:

140-31-8

Molecular formula:

C6H15N3

IUPAC Name:

1-(2-Aminoethyl) piperazine

Test material form:

liquid

Details on test material:

Name of the test chemical: 1-(2-Aminoethyl) piperazine
Molecular Formula:C6H15N3
Molecular Weight: 129.206 g/mol
InChI: 1S/C6H15N3/c7-1-4-9-5-2-8-3-6-9/h8H,1-7H2
Substance Type: Organic
Physical State: Liquid

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

Details on test animal
TEST ANIMALS
- Source: HRP Inc. (Denver, PA).
- Age at study initiation: 5-7 weeks old
- Weight at study initiation: 278-444 grams

Study design: in vivo (non-LLNA)

Induction

Route:

intradermal and epicutaneous

Vehicle:

not specified

Concentration / amount:

0.1ml intradermal induction injections containing 50% (v/v) Freunds complete adjuvant (FCA) water emulsion, 5% test material, and the test material in FCA/water emulsion or FCA/water emulsion.
Epicutaneous induction - 0.2 ml of 50% piperazine

Adequacy of induction:

other: The concentration that produced only local necrosis (i.e., no extensive necrosis or ulceration) was used for the intradermal induction. The highest concentration that produced only mild irritation was considered for the epicutaneous induction.

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

not specified

Concentration / amount:

25%

Day(s)/duration:

24 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10 male and 10 female guinea pigs

Details on study design:

RANGE FINDING TESTS: Range finding studies were conducted to select the appropriate concentration for induction and challenge exposures.
Animals were inspected 24 and 48 h after dosing for signs of necrosis and ulceration. The concentration that produced only local necrosis (i.e., no extensive necrosis or ulceration) was used for the intradermal induction. The highest concentration that produced only mild irritation was appropriate for the epicutaneous induction, and the highest concentration that did not produce irritation was used for the epicutaneous challenge.
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: single
- Exposure period: epicutaneous induction – 48hours
- Test groups: 20 guinea pigs
- Control group: no data
- Site: shoulder
- Frequency of applications: single
- Duration: epicutaneous induction – 48 hours
- Concentrations: 0.1ml intradermal induction injections containing 50% (v/v) Freunds complete adjuvant (FCA) water emulsion, 5% test material, and the test material in FCA/water emulsion or FCA/water emulsion.
Epicutaneous induction - 0.2 ml of 50% test chemical

B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day(s) of challenge: single
- Exposure period:24 hours
- Test groups: 20 guinea pigs
- Control group:
- Site: untreated site (right flank)
- Concentrations: 25% test chemical
- Evaluation (hr after challenge): The sites inspected for signs of irritation 24-48 h after removal of the occlusive dressings

OTHER: Irritation control animals, five male and five female guinea pigs, received the same challenge procedures as in the definitive sensitization study, but did not have preceding intradermal and/or epicutaneous induction procedures. This allowed differentiation between primary skin irritation due to the test material, and those produced by a hypersensitivity reaction.

Grading of Irritation:
Skin responses were evaluated according to a scale as follows:
Evaluation of Skin Responses:
No reaction = 0
Very slight (barely perceptible) erythema, usually non-confluent= 0.5
Slight (well-defined) erythema, usually confluent = 1
Moderate erythema = 2
Severe erythema, with or without edema, necrosis, or eschar formation = 3
Edema and necrosis or eschar formation was noted when present. Erythema or edema at the challenge site of any of the observations greater than that seen in the irritation control animals was considered an allergic response. In general, scores of 1 or greater were considered clearly indicative of sensitization. Scores of 0.5 were considered equivocal, although a high percentage of 0.5 scores with no response in irritation control animals would be considered suggestive of sensitization.
To compare the relative sensitizing potencies, the responses were normalized as follows:
normalized response = %response / [(epicutaneous induction concentration)*(challenge concentration)]

Challenge controls:

no data available

Results and discussion

In vivo (non-LLNA)

Results

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25% challenge concentration

Total no. in group:

20

Clinical observations:

1-(2-Aminoethyl) piperazine appeared to be moderate sensitizer based on the normalized response

Remarks on result:

positive indication of skin sensitisation

Any other information on results incl. tables

Table:Skin Sensitization Responses Tested with the Guinea Pig Maximization Protocol

 

Test chemical	Concentration(%) used for			Skin sensitization response^a
	Intradermal induction	Epicutaneous induction	Epicutaneous challenge	Percentage	Normalized
140 -31 -8	5	50	25	25	0.020

a- Percentage of animal in the group (n=20) showing a positive skin response with score≤l.

b- n=19.

Normalized response=response/[(epicutaneous induction conc.)*(challenge conc.)]

Applicant's summary and conclusion

Interpretation of results:

other: Sensitizing

Conclusions:

The skin sensitization response for the test chemical was 2 5% when tested in 19 guinea pigs. The normalized skin sensitization response was 0.020.
The test chemical appeared to be moderate sensitizer based on the normalized response
Hence, the test chemical can be considered to be sensitizer to the skin of Dunkin Hartley Haz:(DH)fBR albino male and female guinea pigs.

Executive summary:

The dermal sensation potential of the test chemical was evaluated in guinea pigs following the method described by Magnusson and Kligman. 10 male and 10 female Dunkin Hartley Haz:(DH)fBR albino male and female guinea pigs weighing 278-444 grams were used for the study.Range finding studies were conducted to select the appropriate concentration for induction and challenge exposures.

Animals were inspected 24 and 48 h after dosing for signs of necrosis and ulceration. The concentration that produced only local necrosis (i.e., no extensive necrosis or ulceration) was used for the intradermal induction. The highest concentration that produced only mild irritation was appropriate for the epicutaneous induction, and the highest concentration that did not produce irritation was used for the epicutaneous challenge.

The concentrations obtained for the test chemical were – 5% intradermal induction, 50% epicutaneous induction,25% epicutaneous challenge. In the main sensitization study,0.1ml intradermal induction injections containing 50% (v/v) Freunds complete adjuvant (FCA) water emulsion, 5% test material, and the test material in FCA/water emulsion or FCA/water emulsion were injected on 2 sites each of the clipped shoulder skin of guinea pigs. After 7 days of intradermal induction, epicutaneous inductions were performed. 0.2 ml of 50% test chemical was applied in saturation to a 2*4cm filter paper, which was then placed on the test site and secured with a tape. The patches were left in place for 48 hours and after removal of the patches, the skin was wiped free of the excess test material. During the challenge phase, 25% test chemical was soaked in 2*2 cm filter paper squares was applied to a previously untreated site(right flank) 14 days after epicutaneous induction. Patches were left in place for 24 hours, and the sites were inspected for signs of irritation and scored 24 -48 hours after removal of patches.

The skin sensitization response for the test chemical was 2 5% when tested in 19 guinea pigs. The normalized skin sensitization response was 0.020.

The test chemical appeared to be moderate sensitizer based on the normalized response

Hence, the test chemical can be considered to be sensitizer to the skin of Dunkin Hartley Haz:(DH)fBR albino male and female guinea pigs.