1-Hexanaminium, 6-bromo-N,N,N-trimethyl-, bromide (1:1)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

not specified

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

6-Bromhexyltrimethylammoniumbromide

IUPAC Name:

6-Bromhexyltrimethylammoniumbromide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Name: Monoquat
Chemical name: 6-Bromhexyltrimethylammoniumbromid
Molecular formula: C9H21NBr2
CAS No.: 32765-81-4
Supplier: Sponsor
Batch No.: HRHM 002
Melting point: 120 °C
Appearance: White crystalline powder
Solubility in water: Soluble
Conditions of storage: In the dark, may be used under light

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

Species, strain: Guinea pigs, Dunkin Hartley, HsdPoc:DH.
Supplier: Harlan Winkelmann, Gartenstrasse 27, D-33178 Borchen.
Sex, specification: Female healthy young adult and non pregnant animals.
Age of the animals: Approx. 5-7 weeks at the first application.
Weight range of the animals: 281 g to 361 g at the first application.
Number of the animals in the main study: 10 animals for the test substance group; 5 animals for the control group.

Animal maintenance:
Room temperature: Average of 21.9 °C
Relative humidity: Average of 50.5 %
Air exchange: Approx. 12/h.
Light: Only artificial light from 6.00 a.m. to 6.00 p.m.
Cages: Until Day 0: Makrolon cages type III (23 cm x 39 cm bottom area, 18 cm height) with wire mesh lids, Single caging.
From Day 0: group caging in plastic Containers (46 cm x 105 cm x 36 cm), partly shaded, 6 (control group) or 11 (test substance group) animals per Container.
Feed: Altromin Maintenance Diet No. 3122, rich in crude fiber, ad libitum, offered in stainless steel Containers. Analysis of the feed for ingredients and contaminants are performed randomly by Altromin GmbH, D-32791 Lage.
Bedding material: Wood Chips (aspen) from Fa. ABEDD Dominik Mayr KEG, A-8580 Köflach. Reduction of microorganisms by autoclaving.
Water: Tap water offered in Makrolon bottles with stainless steel canules ad libitum.
Identification of the animals: Numbers tattooed in the pinna of the right ear.
Acclimatisation: 5 days.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Induction 2

Concentration / amount:

0.1 mL of 1 % w/v in physiological saline for intradermal induction.
0.5 mL of 50 % w/v in physiological saline for epicutaneous exposure.
0.5 mL of 50 % w/v in deionised water for epicutaneous exposure.
Concentrations were selected after dose finding studies.

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Challenge 2

Concentration / amount:

0.1 mL of 1 % w/v in physiological saline for intradermal induction.
0.5 mL of 50 % w/v in physiological saline for epicutaneous exposure.
0.5 mL of 50 % w/v in deionised water for epicutaneous exposure.
Concentrations were selected after dose finding studies.

No. of animals per dose:

10 animals for the test substance group
5 animals for the control group

Details on study design:

Test principle
First induction exposure: intradermal injections of the test substance, of Freunds Complete Adjuvant FCA (to enhance a possible sensitisation) and of the test substance diluted with FCA. Application site was an area of approx. 2 cm x 4 cm in the interscapular region.
Second induction exposure: epicutaneous application of the test substance to the sites of the intradermal injections.
Challenge exposure: epicutaneous application of the test substance to the left flanks and application of the vehicle to the right flanks of all animals.

Time schedule
Day 0: removal of hair, recording of body weight, intradermal induction exposure.
Day 1: examination of injection sites.
Day 6: removal of hair, treatment with n-dodecylsulfate, sodium salt.
Day 7: removal of hair, epicutaneous induction exposure.
Day 9: end of the epicutaneous induction exposure.
Day 10: skin examination.
Day 20: removal of hair, epicutaneous challenge exposure.
Day 21: end of the epicutaneous challenge period.
Day 22: approximately 21 hours after removing the patch cleaning of the challenge area, approximately 3 hours later skin examination.

Intradermal induction exposure
The application Site for all injections was an area of about 2 cm x 4 cm in the interscapular region. In each of the injection rows two intradermal injections were made side by side.

Epicutaneous induction exposure
The test substance concentration of 50 % (w/v) in physiological saline did not cause remarkable skin irritations in the preliminary test. According to B. Magnusson and A.M. Kligman, all animals of both groups were therefore pretreated with a formulation of n-dodecylsulfate, sodium salt, 10 % (w/w) in white petrolatum, one day before the epicutaneous induction exposure. About 0.5 g per animal was gently applied with a plastic spatula onto the appropriate area to give a slight local hyperaemia. The area was left uncovered.
The test substance was dissolved in physiological saline.
After clipping the hair test patches, about 2 cm x 4 cm, were soaked with the test substance solution (test substance group) and with physiological saline (control group), respectively, and were applied to the area of the intradermal injection. They were fixed with a strip of non-irritating tape. The area of administration was then covered occlusively with aluminium foil and finally fixed with "Fixomull® Stretch".
0.5 mL of test substance formulation or 0.5 mL physiological saline were applied to each animal. The exposure time was 48 hours.

Challenge exposure
The test substance was dissolved in deionised water.
After clipping the hair test patches, about 2 cm x 2 cm, were soaked with the test substance solution, and were applied to the left flanks of all animals of both groups. Test patches of the same size, soaked with deionised water, were applied to the right flanks of all animals. Mode of fixation was the same as for the epicutaneous induction exposure.
0.5 mL of test substance formulation and 0.5 mL of deionised water were applied to each animal. The exposure time was 24 hours.

Challenge controls:

Challenge exposure: epicutaneous application of the test substance to the left flanks and application of the vehicle to the right flanks of all animals.

Positive control substance(s):

yes

Remarks:

Periodic checks with hexyl cinnamic aldehyde.

Results and discussion

Positive control results:

7 out of 10 animals were sensitised by the positive control substance at the last check.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

50 %

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

50 %

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

50 %

No. with + reactions:

8

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 8.0. Total no. in groups: 10.0.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

50 %

No. with + reactions:

8

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 8.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Mortality: All animals survived till the end of the study.

Body weight: There were no significant differences in the mean body weights between the animals of the test substance group and those of the control group on Days 0 and 23.

General observations: No unexpected abnormal behaviour or clinical signs were detected during the experiment in the animals.

Skin reactions after the challenge exposure:

Negative control group:

Vehicle site: No positive skin reaction in any animal at any reading time.

Test substance site: No positive skin reaction in any animal at any reading time.

Test substance group:

Vehicle site: No positive skin reaction in any animal at any reading time.

Test substance site: 8/10 animals had very slight to severe erythema and/or oedema 24 and/or 48 hours after the end of the challenge exposure. In animal Nos. 11 and 13 the severe erythema and/or oedema were attended additionally by eschars 48 hours after the end of the challenge exposure.

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Monoquat is considered to be a skin sensitizer in the guinea pig maximisation test.

Executive summary:

A guinea pig maximisation test according to OECD 406 method was performed. Monoquat is considered to be a skin sensitizer in the this test, as 8 out of 10 animals were sensitised.