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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:
2008-10-08 to 2009-04-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-compliant study under GLP without deviations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): NLP #15 (2,2',2''-Nitrilotriethanol, propoxylated)
- Molecular weight (if other than submission substance): 340 g/mol
- Physical state: liquid
- Analytical purity: 100 %
- Purity test date: 2008-08-28
- Lot/batch No.: 2224R01
- Expiration date of the lot/batch: 26-Feb-2009
- Storage condition of test material: at room temperature in the dark
- Stability: Stable in water for at least 6 months if stored in the dark
or under diffuse light and/or in limited access of air;
unknown in a 1:1 (v/v) mixture of FCA/physiological
saline; is excluded from the statement of compliance.

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS Albino Dunkin Hartley Guinea Pig, CRL:(HA)BR, SPF (Specific Pathogen Free)
- Source: Charles River, Kisslegg, Germany
- Age at study initiation: 9-11 weeks
- Weight at study initiation: 538-704 g
- Housing:Individually in Makrolon type-4 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard guinea pig breeding / maintenance diet (Provimi Kliba AG, Kaiseraugst, Switzerland) ad lib.
-Water (e.g. ad libitum):Community tap water ad lib.
- Acclimation period: under laboratory conditions


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 3
- Humidity (%): 30-70
- Air changes (per hr): at least 15/h
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 2008-11-12 to 2008-12-09

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
5% for intradermal induction, 100% for epidermal induction;
75% for epidermal challenge
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
5% for intradermal induction, 100% for epidermal induction;
75% for epidermal challenge
No. of animals per dose:
10 control animals and 20 test animals
Details on study design:
RANGE FINDING TESTS:
Intradermal pretest (together with Freund's) in two animals, 75-50-25-15-10 and 5 %. Dermal reactions were assessed 24 hours later based on the results, the test item concentration of 5 % (w/w) was selected for intradermal induction in the main study.
Epidermal pretest (after Freund's) in two animals, 100-75-50 and 25 %. Based on the results, the test item concentration of 75 % (w/w) was selected for epidermal challenge and 100% as lowest irritating concentration for epidermal induction.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 1x intradermal, 1 x epicutaneous
- Exposure period: intradermal-continuous, epicutaneous: 48 h
Test group:
intradermal induction:
in parallel intradermal injection of Freund's complete adjuvant, test item (5% ) in purified water, Freund's/test item (5%) 1:1
epidermal induction:
100% test item, occlusive conditions, 48 h
- Control group:
intradermal induction:
in parallel intradermal injection of Freund's complete adjuvant, purified water, Freund's/purified 1:1
epidermal induction:
purified water, occlusive conditions, 48 h
- Site: dorsal skin from the scapular region (intradermal and epicutaneous)
- Frequency of applications: once



B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day(s) of challenge: day 22
- Exposure period: 24 h
- Test groups: test item (75%) / water
- Control group: test item (75%) / water
- Site: left flank
- Evaluation (hr after challenge): 24 h, after depilation
Challenge controls:
No re-challenge
Positive control substance(s):
yes
Remarks:
Alpha-Hexylcinnamaldehyde

Study design: in vivo (LLNA)

Statistics:
Descriptive statistics (means and standard deviations) were calculated for body weights. No
inferential statistics were used

Results and discussion

Positive control results:
90% of positive responders in test group, 0% in control group

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water
No. with + reactions:
6
Total no. in group:
10
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the control group
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water . No with. + reactions: 6.0. Total no. in groups: 10.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the control group.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the control group
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water . No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the control group.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
purified water only
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the control group
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: purified water only . No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the control group.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
purified water only
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the control group
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: purified water only . No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the control group.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water
No. with + reactions:
8
Total no. in group:
20
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the test group
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water . No with. + reactions: 8.0. Total no. in groups: 20.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the test group.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the test group
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: NLP#15 (2,2’,2’’-Nitrilotriethanol, propoxylated), 75 % in purified water . No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the test group.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
purified water only
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the test group.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: purified water only . No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the test group..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
purified water only
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No systemic toxicity and/or deaths were noted in the guinea pigs of the test group.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: purified water only . No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No systemic toxicity and/or deaths were noted in the guinea pigs of the test group..

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
No skin sensitizing potential was identified in the guinea pig Maximization-test with NLP#15
Executive summary:

The skin sensitizing potential of the test item NLP#15 was investigated in a guinea pig maximisation test according to OECD 406. Male guinea pigs were treated intradermally (together with Freund's adjuvant) and epicutaneously (lowest irritating concentration) with the test item. 21 days after first administration, the test item was applied epicutaneously at the highest non-irritating concentration

The local reactions recorded in the eight test animals (40 %) after the challenge were considered to be of irritant nature. This interpretation is supported by the fact that the reactions faded at the 48-hour reading and that 60 % of the control animals also showed a local erythema at the 24 -hour reading.

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, NLP #15 (2,2’,2’’-Nitrilotriethanol, propoxylated) does not have to be classified and labelled as a skin sensitizer.