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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 December 1993 - 25 January 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study from 1994

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Product name: Amgard V6
Appearance: very pale straw coloured viscous liquid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: David Hall Limited, Burton-on-Trent, UK
- Age at study initiation: 8 - 12 weeks old
- Weight at study initiation: 355 - 450 grams
- Housing: in groups of up to three, in solid floor polypropylene cages
- Diet: ad libitum (Guinea Pig FDI Diet)
- Water: ad libitum
- Acclimation period: minimum five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 22
- Humidity (%): 42 - 65
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12h/12h

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Epicutaneous exposure undiluted (no vehicle), for intradermal exposure Acetone 6% in Arachis oil used as vehicle
Concentration / amount:
- Intradermal induction: sighting test with 1 and 5% w/v of the test substance in 6% acetone v/v in arachis oil, main test only with 5% w/v
- Epicutaneous induction: sighting test with 25, 50, 75 and 100 % v/v of the test material in acetone, main study only with 100 % v/v (undiluted)
- Epicutaneous challenge: sighting test with 25, 50, 75 and 100 % v/v of the test material in acetone, main study only with 75 and 100 % v/v (undiluted)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Epicutaneous exposure undiluted (no vehicle), for intradermal exposure Acetone 6% in Arachis oil used as vehicle
Concentration / amount:
- Intradermal induction: sighting test with 1 and 5% w/v of the test substance in 6% acetone v/v in arachis oil, main test only with 5% w/v
- Epicutaneous induction: sighting test with 25, 50, 75 and 100 % v/v of the test material in acetone, main study only with 100 % v/v (undiluted)
- Epicutaneous challenge: sighting test with 25, 50, 75 and 100 % v/v of the test material in acetone, main study only with 75 and 100 % v/v (undiluted)
No. of animals per dose:
- Sighting tests: 3 x 2 animals, to select concentrations for the main study
- Main study: 20 animals for the treatment, 10 as control
Details on study design:
MAIN STUDY
- No. of exposures: 3 injections (intradermal) at Day 0, epicutaneous application at Day 7, epicutaneous challenge at Day 21
- Site: both intradermal and epicutaneous induction on skin site on right shoulder, epicutaneous challenge on right flank
- Exposure period: epicutaneous induction with 48 hours exposure, challenge with 24 hours exposure
- Frequency of applications: 1 intradermal, 2 epicutaneous (induction and challenge)
- Duration: 24 days (total)
- Evaluation: 24 and 48 hours after intradermal application, 1 and 24 hours after epicutaneous induction and 24, 48 and 72 hours after epicutaneous challenge
- Sodium laurylsulfate pretreatment (0.5 ml of a 10% solution in petrolatum) was used to enhance the irritant reaction on day 6 after the intradermal
induction.
Challenge controls:
Vehicle applied on left flank, application similarl to treatment group
Positive control substance(s):
yes
Remarks:
recent historical control data with 2,4-dinitrochlorobenzene, alpha-hexycinnamaldehyde and 2-mercaptbenzthiazole were provided

Results and discussion

Positive control results:
2,4-dichloronitrobenzene: intradermal induction conc. 0.5% in arachis oil, topical induction: 0.75% in abs. ethanol, challenge: 0.25% in abs. ethanol.:
9/10 positive
alpha-hexycinnamaldehyde (85%): intradermal induction conc. 25% in arachis oil, topical induction: 100%, challenge: 100% and 75% in arachis oil.:
2 studies: one 3/10 positive, one 7/10 positive
2-mercaptobenzthiazole: intradermal induction conc. 5% in arachis oil, topical induction: 50% in archis oil, challenge: 50 and 25% in arachis oil:
8/10 positive

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
other: Intradermal induction
Hours after challenge:
24
Group:
test group
Dose level:
5% w/v
No. with + reactions:
20
Total no. in group:
20
Remarks on result:
other: Reading:
Reading:
other: Intradermanl induction
Hours after challenge:
48
Group:
test group
Dose level:
5% w/v
No. with + reactions:
20
Total no. in group:
20
Remarks on result:
other: Reading:
Reading:
other: Intradermal induction
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Intradermal induction
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous induction
Hours after challenge:
1
Group:
test group
Dose level:
undiluted
No. with + reactions:
16
Total no. in group:
20
Clinical observations:
2 animals found dead, one at Day 8 and one at Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous induction
Hours after challenge:
24
Group:
test group
Dose level:
undiluted
No. with + reactions:
5
Total no. in group:
20
Clinical observations:
2 animals found dead, one at Day 8 and one at Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous induction
Hours after challenge:
1
Group:
negative control
Dose level:
0
No. with + reactions:
7
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous induction
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
24
Group:
test group
Dose level:
undiluted
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
2 animals found dead, one at Day 8 and one at Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
24
Group:
test group
Dose level:
75% v/v
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
2 animals found dead, one at Day 8 and one at Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
48
Group:
test group
Dose level:
undiluted
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
2 animals found dead, one on Day 8 and one on Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
48
Group:
test group
Dose level:
75% v/v
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
2 animals found dead, one on Day 8 and one on Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
72
Group:
test group
Dose level:
undiluted
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
2 animals found dead, one on Day 8 and one on Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
72
Group:
test group
Dose level:
75% v/v
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
2 animals found dead, one on Day 8 and one on Day 9
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Epicutaneous challenge
Hours after challenge:
72
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: Intradermal induction
Hours after challenge:
24
Group:
positive control
Dose level:
5% w/w
No. with + reactions:
0
Total no. in group:
10

Any other information on results incl. tables

- Intradermal induction: very slight to well-defined erythema (grades 1 -2) at 24 and 48 hours in all tst animals, no reactions in control group.

- Epicutaneous induction: very slight erythema (grade 1) with or without very slight oedema (grade 1) noted at 16 test animals after 1 hour, in 5 animals also after 24 hours. Very slight erythema (grade 1) noted in 7 control animals after 1 hour, very slight oedema (grade 1) noted in 1 control group animal after 1 hour.

- Epicutaneous challenge undiluted test substance: very slight erythema (grade 1) noted in 2 test animals after 24 hours, in 1 animal after 48 hours and in none of the animals after 72 hours. No skin reactions in control group at any time.

-Epicutaneous challenge 75% in acetone: No skin reactions were observed in all the challenge sites of test animals and controls at all time points.

- 17% of the test animals (3/18) showed a positive skin reaction at challenge. According to the test guideline this result can be regarded as negative as less than 30% of the animals showed a positive challenge reaction. The test material is therefore not a skin sensitizer under the conditions of this study.

2 animals died during the study for undefined reasons.

Body weight gains of the test group were comparable to those observed in the negative control group.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The substance is not considered a skin sensitizer in this study.
Executive summary:

In a dermal sensitization study, 2,2-bis(chloromethyl)trimethylene bis(bis(2-chloroethyl)phosphate) was tested in female albino Dunkin-Hartley guinea pigs according to OECD Test Guideline No.406 (guinea pig maximisation test). At challenge with the undiluted test substance 3 of 18 animals (17%) showed a slight skin reaction. None reacted positively in the group challenged with 75% of the test substance in acetone. According to the test guideline the substance is not considered a skin sensitizer in this study. Two test animals died during the study for undefined reasons. Body weight gains of the test animals were comparable to those of the negative controls.