Poly(oxy-1,2-ethanediyl), alpha-[2-[bis(2-aminoethyl)methylammonion)ethyl]-, omega-hydroxy, N,N'-di-Limnanthes alba seed oil acyl derivatives, methyl sulfates (salts)

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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

17 July 1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

November 2000

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Poly(oxy-1,2-ethanediyl), alpha-[2-[bis(2-aminoethyl)methylammonion)ethyl]-, omega-hydroxy, N,N'-di-Limnanthes alba seed oil acyl derivatives, methyl sulfates (salts)

EC Number:

629-704-1

Cas Number:

226995-92-2

Molecular formula:

The substance is a UVCB - no specific molecular weight or formula is available.

IUPAC Name:

Poly(oxy-1,2-ethanediyl), alpha-[2-[bis(2-aminoethyl)methylammonion)ethyl]-, omega-hydroxy, N,N'-di-Limnanthes alba seed oil acyl derivatives, methyl sulfates (salts)

Test material form:

liquid

Specific details on test material used for the study:

Batch number: CW-4896
Storage conditions: at room temperature
Purity: 85.9% actives
Expiry date: 2 years

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

Nulliparous and non-pregnant.
On the first day of treatment animals were approximately 9 weeks old and had a mean body weight of 20.5 ± 1.0g.
Acclimitisation: 5 days

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

Preliminary test: 5, 10, 25 and 50% concentration.
Main test - first experiment: 0, 1, 2.5 and 5%.
Main test - second experiment: 0, 0.1, 0.5 and 1%.

No. of animals per dose:

Preliminary test: 4 animals
Main test: 4 animals/dose

Positive control substance(s):

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

Results and discussion

Positive control results:

In the positive control group given HCA at the concentration of 25%, a moderate increase in cellularity and a stimulation index exceeding the threshold value of 3 (First experiment: SI = 4.62; Second experiment: SI = 6.97) was noted. The study was therefore considered to be valid.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

17.36

Test group / Remarks:

1%

Results 2

Key result

Parameter:

SI

Value:

47.89

Test group / Remarks:

2.5%

Results 3

Key result

Parameter:

SI

Value:

6.43

Test group / Remarks:

5%

Results 4

Key result

Parameter:

SI

Value:

2.03

Test group / Remarks:

0.1%

Results 5

Key result

Parameter:

SI

Value:

6.98

Test group / Remarks:

0.5%

Results 6

Key result

Parameter:

SI

Value:

14.58

Test group / Remarks:

1%

Cellular proliferation data / Observations:

Preliminary test:
The substance was an irritant at all concentrations tested.

Main test:
No clinical signs and no mortality was observed during the study.
The body weight gain of the treated animals was similar to that of the controls.
A slight to moderate erythema (grades 1-3) was noted in all animals given the concentrations of 2.5% and 5%. Dryness of the skin was recorded at the concentrations 1, 2.5 and 5% and crusts at the concentration of 5%. These skin reactions were associated with increase in ear thickness at the concentrations of 5, 2.5, 1 and 0.5%.
No cutaneous reactions and no increase in ear thickness were observed in the animals given the concentration of 0.1%.
A significant lymphoproliferation was noted in the treated groups given the concentration of 5, 2.5, 1 and 0.5%. However, as this lymphoproliferation was not dose-dependent and as it could be due to the sole irritant effect observed at these same concentrations, it was not attributed to delayed contact hypersensitivity. No lymphoproliferation was noted at the concentration of 0.1%.

Any other information on results incl. tables

Study results - First Experiment

Concentration (%)	0	1	2.5	5
Viability (%)	91.67	89.89	80.32	91.03
Cellularity index	-	9.09	5.66	9.68
Stimulation Index	-	17.36	47.89	6.43
Increase in ear thickness (% between day 1 -6)	0	23.3	68.63	83.5

Study results: Second Experiment

Concentration (%)	0	0.1	0.5	1
Viability (%)	95.93	95.14	95.14	96.98
Cellularity index	-	1.07	2.85	4.67
Stimulation Index	-	2.03	6.98	14.58
Increase in ear thickness (% between day 1 -6)	3.06	7.14	23.71	42.27

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The substance induces a lymphoproliferation which is most probably not attributable to delayed contact hypersensitivity in the murine LLNA.

Executive summary:

This study was undertaken to evaluate the potential of the substance to induce delayed contact hypersensitivity using the murine LLNA. Evaluation of irritation was also conducted in parallel by measurement of ear thickness. A preliminary test was conducted in order to define the concentration of the substance to be usedin the main test. There were two experiments conducted in the main test. The first experiment employed substance concentrations of 1, 2.5 and 5% and the second experiment employed 0.1, 0.5 and 1%. The second experiment was included becuase irritation occurred at all concentrations in the first experiment. In both experiments a negative group that received the vehicle only and a positive group that received alph hexyl cinnamaldehyde (HCA) only was also included. Four females of CBA/J strain mice were included per dose group. During the induction phase the substance was applied over the ears (25 µL/ear) for three consecutive days. After 2 days of resting, the proliferation of lymphocytes in the lymph node draining the application site was measured by incoporation of tritiated methyl thymidine (day 6). The obtained values were used to calclulate the Stimulation Index (SI). The substance was an irritant at all concentrations tested in the preliminary test. In the main test there were no clinical signs and no mortality observed.

The body weight gain of the treated animals was similar to that of the controls. A slight to moderate erythema (grades 1-3) was noted in all animals given the concentrations of 2.5% and 5%. Dryness of the skin was recorded at the concentrations 1, 2.5 and 5% and crusts at the concentration of 5%. These skin reactions were associated with increase in ear thickness at the concentrations of 5, 2.5, 1 and 0.5%. No cutaneous reactions and no increase in ear thickness were observed in the animals given the concentration of 0.1%. A significant lymphoproliferation was noted in the treated groups given the concentration of 5, 2.5, 1 and 0.5%. However, as this lymphoproliferation was not dose-dependent and as it could be due to the sole irritant effect observed at these same concentrations, it was not attributed to delayed contact hypersensitivity. No lymphoproliferation was noted at the concentration of 0.1%. In conclusion, the substance induces a lymphoproliferation which is most probably not attributable to delayed contact hypersensitivity in the murine LLNA.