3-chloropropene

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: - guideline compliant study - GLP compliant

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

3-chloropropene

EC Number:

203-457-6

EC Name:

3-chloropropene

Cas Number:

107-05-1

Molecular formula:

C3H5Cl

IUPAC Name:

3-chloroprop-1-ene

Details on test material:

Sponsor's identification: Allyl chloride
Description: clear colourless liquid
Analytical purity: 98.7%
Impurities: water 18 mg/kg
Purity test date: 18 November 2009
Batch number: RBA091118A
Expiry date: 27 November 2011
Stability under test conditions: stable under recommended storage conditions
Date received: 27 November 2009
Storage conditions: approximately 4°C in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Bicester, Oxon, UK
- Age at study initiation: 8 - 12 wks
- Weight at study initiation: 15 to 23 g
- Housing: individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes
- Diet (e.g. ad libitum): ad libitum, 2014 Teklad Global Rodent diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK
- Water (e.g. ad libitum): ad libitum, tap water
- Acclimation period: > 5 d


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25
- Humidity (%): 30 to 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 2010-01-13 To: 2010-01-27

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

25, 50 % (v/v) in olive oil and 100 % (undiluted)

No. of animals per dose:

5

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: completely soluble in the vehicle
- Irritation: The preliminary screening test suggested that the test material would not produce systemic toxicity or excessive local irritation at the highest suitable concentration.
- Lymph node proliferation response: not assessed


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: random
- Criteria used to consider a positive response: The test material will be regarded as a sensitiser if at least one concentration of the test material results in a threefold or greater increase in 3HTdR incorporation compared to control values.


TREATMENT PREPARATION AND ADMINISTRATION: animals, were treated with 50 pl (25 pl per ear) of the undiluted test material or the test material as a solution in acetone/olive oil 4:1 at concentrations of 50% or 25% v/v. A further group of five animals was treated with acetone/olive oil 4:1 alone. A concurrent positive control test, using a group of five animals, was also performed with the known sensitiser, a-Hexylcinnamaldehyde tech., 85%, at a concentration of 15% v/v in acetone/olive oil 4:1.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Data was processed to give group mean values for disintegrations per minute and standard deviations where appropriate. Individual and group mean disintegrations per minute values were assessed for dose response relationships by analysis of homogeneity of variance followed by one way analysis of variance (ANOVA). In the event of a significant result from the ANOVA, pairwise comparisons were performed between control and treated groups. For homogenous datasets Dunnett's Multiple Comparison test was used and for non-homogenous datasets Dunnett's T3 Multiple Comparison Method was used.

Results and discussion

Positive control results:

a-Hexylcinnamaldehyde tech., 85%, at a concentration of 15% v/v in acetone/olive oil 4:1: stimulation index: 6.21, Positive

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: < 3 for all treatment groups, > 3 for the positive test substance, see table 1

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see table 1

Any other information on results incl. tables

- table 1: Individual Disintegrations per Minute and Stimulation Indices

Treatment Group	Animal Number	dpm/ Animal a	Mean dpm/Animal (Standard Deviation)	Stimulation Index b	Result
Vehicle acetone/olive oil 4:1	1 -1	1668.60	1655. 95 (±417.16)	N/A	N/A
	1-2	1290.06
	1-3	2285.84
	1-4	1769.67
	1-5	1265.60
Test Material 25% v/v in acetone/olive oil 4:1	2-1	938.63	1293.35 (±358.65)	0.78	Negative
	2-2	1558.52
	2-3	1699.85
	2-4	1361.70
	2-5	908.03
Test Material 50% v/v in acetone/olive oil 4:1	3-1	1484.54	1246.26 (±378.07)	0.75	Negative
	3-2	1252.86
	3-3	1384.65
	3-4	595.10
	3-5	1514.15
Test Material 100%	4-1	5363.54	3255 21 . (±1380.91)	1.97	Negative
	4-2	3626.75
	4-3	1772.00
	4-4	3174.02
	4-5	2339.74
Positive Control Material 15% v/v in acetone/olive oil 4:1	5-1	6359.41	10278.58* (±3233.93)	6.21	Positive
	5-2	14875.75
	5-3	8667.83
	5-4	9670.29
	5-5	11819.63

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

3-chloropropene was tested in a local lymphnode assay according to OECD TG 429.
The test material was considered to be a non-sensitiser under the conditions of the test.
a-Hexylcinnamaldehyde, tech., 85% gave a Stimulation Index of greater than 3 when tested at a concentration of 15% v/v in acetone/olive oil 4:1

Executive summary:

The present study (Sanders 2010) was performed to assess the skin sensitisation potential of the test material 3 -chloropropene in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear. The method was designed to meet the requirements of the following:

OECD Guideline for the Testing of Chemicals No. 429 "Skin Sensitisation: Local Lymph Node Assay" (adopted 24 April 2002) Method B42 Skin Sensitisation (Local Lymph Node Assay) of Commission Regulation (EC) No. 440/2008 Methods.

Following a preliminary screening test in which no clinical signs of toxicity were noted at a concentration of 100%, this concentration was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of five animals, were treated with 50 µl (25 µl per ear) of the undiluted test material or the test material as a solution in acetone/olive oil 4:1 at concentrations of 50% or 25% v/v. A further group of five animals was treated with acetone/olive oil 4:1 alone. A concurrent positive control test, using a group of five animals, was also performed with the known sensitiser, a-Hexylcinnamaldehyde tech., 85%, at a concentration of 15% v/v in acetone/olive oil 4:1. Results. The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

Treatment Group	Concentration	Stimulation Index	Result
Test Material	25% v/v in acetone/olive oil 4:1	0.78	Negative
	50% v/v in acetone/olive oil 4:1	0.75	Negative
	100%	1.97	Negative
Positive Control Material	15% v/v in acetone/olive oil 4:1	6.21	Positive

The test material was considered to be a non-sensitiser under the conditions of the test. a-Hexylcinnamaldehyde, tech., 85% gave a Stimulation Index of greater than 3 when tested at a concentration of 15% v/v in acetone/olive oil 4:1 thereby validating the test system.