Docosyltrimethylammonium methyl sulphate

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008-05-07 till 2008-06-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD TG 406)

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation: 4-5 weeks
- Weight at study initiation: 336 - 345 g
- Housing: Individually in Makrolon type-4 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 guinea pig breeding / maintenance diet, at libidum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum
- Acclimation period: 2008-05-07 to 2008-05-19

ENVIRONMENTAL CONDITIONS
- Temperature (°C): room temperature 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

MAIN STUDY
A: INDUCTION EXPOSURE (TEST GROUP) (intradermal injections and epidermal induction)
Day 1: three intradermal injections/animal (0.1 mL each)
- FCA/physiological saline, 1:1
- 5% test item in purified water
- 5% test item in a 1:1 mixture FCA/physiological saline

Day 8: 0.3 g epicutaneously (50% of test item)

B. CHALLENGE EXPOSURE
Day 22: 0.2 mL of test item (3%) on 5 x 5 cm area on the left flank; 0.2 mL vehicle (purified water) on 5 x 5 cm area on the right flank
Day 23: 3 hrs and 24 hrs after dressing removal skin reactions were checked.

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

MAIN STUDY
A: INDUCTION EXPOSURE (TEST GROUP) (intradermal injections and epidermal induction)
Day 1: three intradermal injections/animal (0.1 mL each)
- FCA/physiological saline, 1:1
- 5% test item in purified water
- 5% test item in a 1:1 mixture FCA/physiological saline

Day 8: 0.3 g epicutaneously (50% of test item)

B. CHALLENGE EXPOSURE
Day 22: 0.2 mL of test item (3%) on 5 x 5 cm area on the left flank; 0.2 mL vehicle (purified water) on 5 x 5 cm area on the right flank
Day 23: 3 hrs and 24 hrs after dressing removal skin reactions were checked.

No. of animals per dose:

5 control animals (females)
10 test animals (females)

Details on study design:

RANGE FINDING TESTS:
A: intradermal injections:
Four intradermal injections (0.1 mL/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline (day 1) (shaved neck of one guinea pig)
Four intradermal injections (0.1 mL/site) at concentrations of A = 15 %, B = 10 % and C = 5 % of the test item in purified water (day 7) (clipped flank of the same guinea pig)

B: epidermal applications:
Four intradermal injections (0.1 mL/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline (day 1) (shaved neck of two guinea pigs)
Epidermal application with the test item at D = 50 %, E = 25 %, F = 5 % and G = 10 % in purified water (day 7), (shaved flank of the same guinea pigs)


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal/epicutaneous)
- Exposure period: injected/48 hrs occlusive
- Test groups: scapular region
- Control group: only vehicle (purified water)
- Frequency of applications: 1 / 1
- Duration: 0 - 8 days
- Concentrations: 5% epidermal; 50% epicutaneous


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: 24 hrs
- Site: left flank (test item); right flank: purified water
- Concentrations: 3% epicutaneous
- Evaluation (hr after challenge): 3 hrs and 24 hrs after dressing removal

Positive control substance(s):

yes

Remarks:

ALPHA-HEXYLCINNAMALDEHYDE

Positive control results:

Skin Effects after epidermal Induction :
Discrete/patchy erythema was observed in all animals at the 24- and 48-hour reading after treatment with the test item at 50 % in purified water.

Skin Effects after Challenge:
No positive/skin reactions were observed in the animals when treated with either purified water only or when treated with the test item at 3 % in purified water.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5% i.d., 50% epic., 3% epic.

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5% i.d., 50% epic., 3% epic.. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: None.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

5% i.d., 50% epic., 3% epic.

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5% i.d., 50% epic., 3% epic.. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the above mentioned findings in an adjuvant sensitization test (M&K-test) in guinea pigs and in accordance to Commission Directive 2001/59/EC, the submission substance does not have to be classified and labelled as a skin sensitizer.

Executive summary:

The sensitization potential of the submission substance was evaluated in guinea-pig according to the Maximization-Test by Magnusson and Kligman.

The intradermal induction of sensitization in the test group was performed in the nuchal region with a 5 % dilution of the test item in purified water and in an emulsion of Freund's Complete Adjuvant (FCA)/physiological saline. The epidermal induction of sensitization was conducted for 48 hours under occlusion with the test item at 50 % in purified water one week after the intradermal induction. The animals of the control group were intradermally induced with purified water and FCA/physiological saline and epidermally induced with purified water under occlusion.

Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test item at 3 % in purified water and purified water alone under occlusive dressing.

Cutaneous reactions were evaluated at 24 and 48 hours after removal of the dressing.

No toxic signs were evident in the guinea pigs of the control or test group. No deaths occurred. No skin reactions were observed after the challenge treatment with the submission substance at 3 % in purified water or purified water alone in neither the control group nor the test group.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

The sensitization potential of the submission substance was evaluated in guinea-pig according to the Maximization-Test by Magnusson and Kligman.

The intradermal induction of sensitization in the test group was performed in the nuchal region with a 5 % dilution of the test item in purified water and in an emulsion of Freund's Complete Adjuvant (FCA)/physiological saline. The epidermal induction of sensitization was conducted for 48 hours under occlusion with the test item at 50 % in purified water one week after the intradermal induction. The animals of the control group were intradermally induced with purified water and FCA/physiological saline and epidermally induced with purified water under occlusion.

Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test item at 3 % in purified water and purified water alone under occlusive dressing.

Cutaneous reactions were evaluated at 24 and 48 hours after removal of the dressing.

No toxic signs were evident in the guinea pigs of the control or test group. No deaths occurred. No skin reactions were observed after the challenge treatment with the test item at 3 % in purified water or purified water alone in neither the control group nor the test group.

A similar supporting substance was tested in a guideline-compliant in vivo skin sensitization test in guinea pigs according to the Buehler method using 20% test item (containing 80% test substance in isopropanol) in ethanol:water 80:20 for epicutaneous induction and 0.8% test item in isopropanol for epicutaneous challenge. None of 20 animals of the treatment group showed a positive skin response after the challenge procedure; also none of 10 control animals showed skin reactions. Based on the results of this study the test item showed no evidence for sensitising properties. The solvent (isopropanol) in the test material is not considered to have influenced the selection of the test concentrations because its skin irritative effects are small compared to those of the test substance which is a surfactant. It is concluded that no higher concentrations of test substance could have been tested if the substance had been used in pure form. Therefore, the study is considered fully adequate to assess the skin sensitizing properties of the supporting substance.

Migrated from Short description of key information:
A guideline study (OECD TG 406) with the submission substance and another guideline study (OECD TG 406) with a similar supporting substance showed no evidence for sensitising properties.

Justification for selection of skin sensitisation endpoint:
Reliable study.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

Migrated from Short description of key information:
Respiratory tract sensitisation:
Substance is not classified for this endpoint. The substance is considered not to exert any sensitizing effects on the respiratory tract.

Justification for classification or non-classification

A guideline study (OECD TG 406) with the submission substance and another guideline study (OECD TG 406) with a similar supporting substance showed no evidence for sensitising properties.

It can reasonably be deduced that the submission substance does not cause respiratory tract sensitization and thus does not have to be classified according to the criteria laid down in the EU Dangerous Substances Directive (67/548/EEC) and in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).