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Administrative data

Description of key information

The test item is considered to be a non-sensitizer under the conditions of a local lymph node assay.

No information is available for respiratory sensitization, however, the substance is a waxy solid with a low vapour pressure and therefore respiratory sensitization is considered unlikely.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was conducted between 20 May 2016 and 08 June 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
22 July 2010
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
EC No. 440/2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
mouse local lymph node assay (LLNA)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sponsor Lot/Batch no.: 5399561P10
- Expiration date of the lot/batch: 25 January 2020
- Purity test date: 25 January 2016


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
Species:
mouse
Strain:
CBA/Ca
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS B.V., Inc, the Netherlands
- Females (if applicable) nulliparous and non-pregnant: yes
- Microbiological status of animals, when known:
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 15 - 23 g
- Housing: Suspended solid floor polypropylene cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: >5 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 ”C
- Humidity (%): 30 - 70%
- Air changes (per hr): >15
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES: From: 20 May 2016 To: 08 June 2016
Vehicle:
propylene glycol
Concentration:
Preliminary Screening: 10% w/w in propylene glycol (maximum attainable concentration
Main Test: 10%, 5% and 2.5% w/w in propylene glycol
No. of animals per dose:
Preliminary screening: 1 mouse
Main test 5 mice /concentration
Details on study design:
PRE-SCREEN TESTS:
- Compound solubility: Maximum attainable concentration = 10% w/w
- Irritation: scored daily
- Systemic toxicity: not determined
- Ear thickness measurements: Between Days 1 and 3 and Days 1 and 6
- Erythema scores: no

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response: Preliminary test: A mean ear thickness increase of equal to or greater than 25% was considered to indicate excessive irritation and limited biological relevance to the endpoint of sensitization.
Main test: The test item will be regarded as a sensitizer if at least one concentration of the test item results in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test item failing to produce a threefold or greater increase in 3HTdR incorporation will be classified as a "non-sensitizer".

TREATMENT PREPARATION AND ADMINISTRATION:
Positive control substance(s):
other: Phenylacetaldehyde
Statistics:
Data was processed to give group mean values for disintegrations per minute and standard deviations where appropriate. Individual and group mean disintegrations per minute values were assessed for dose response relationships. Data was first assessed for suitability by analysis of normality and homogeneity of variance. If the assumptions that the data are both normally distributed and has homogeneity of variances, then parametric one way analysis of variance (ANOVA) and Dunnett’s multiple comparison procedure were used to determine
statistical significance. If the assumptions were not met, non-parametric Kruskal-Wallis Rank Sum and Mann-Whitney U test procedures were used.
Positive control results:
The positive control Phenylacetaldehyde (>90%) gave a Stimulation Index of greater than 3 (15.12) when tested at a concentration of 5% v/v in propylene glycol, thus, demonstrating the sensitivity and reliability of the test system.
Key result
Parameter:
SI
Value:
> 0.92 - < 1.77
Test group / Remarks:
Overall range from all test groups
Key result
Parameter:
SI
Remarks:
5%
Value:
> 0.92 - <= 1.77
Key result
Parameter:
SI
Remarks:
10%
Value:
> 0.92 - <= 1.77
Key result
Parameter:
SI
Remarks:
2.5%
Value:
> 0.92 - <= 1.77
Cellular proliferation data / Observations:

DETAILS ON STIMULATION INDEX CALCULATION The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group

Body weight change of the test animals between Day 1 and Day 6 was comparable to that observed in the corresponding control group animals over the same period.
There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test.

Estimation of the Proliferative Response of Lymph Node Cells - Stimulation Index

 Concentration in propylene glycol  Stimulation Index  Result
 RD 14156 2.5% w/w  0.92  Negative
 RD 14156 5% w/w  1.29  Negative
 RD 14156 10% w/w  1.77  Negative
 Positive control 5% v/v  15.12  Positive
Interpretation of results:
GHS criteria not met
Remarks:
non-sensitizer
Conclusions:
The test item was considered to be a non-sensitizer under the conditions of the test.
The positive control Phenylacetaldehyde (>90%) gave a Stimulation Index of greater than 3 (15.12) when tested at a concentration of 5% v/v in propylene glycol, thus, demonstrating the sensitivity and reliability of the test system.
Executive summary:

A study was performed to assess the skin sensitization potential of the test item in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear.

Following a preliminary screening test in which no clinical signs of toxicity were noted at a maximum attainable concentration of 10% w/w, this concentration was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of five animals, were treated with 50 μL (25 μL per ear) of the test item as a suspension in propylene glycol at concentrations of 10%, 5% or 2.5% w/w. A further group of five animals was treated with propylene glycol alone. A concurrent positive control test, using a group of five animals, was also performed with the known sensitizer, Phenylacetaldehyde (>90%), at a concentration of 5% v/v in propylene glycol.

The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group.

The test item was considered to be a non-sensitizer under the conditions of the test. The positive control Phenylacetaldehyde (>90%) gave a Stimulation Index of greater than 3 (15.12) when tested at a concentration of 5% v/v in propylene glycol, thus, demonstrating the sensitivity and reliability of the test system.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test. Body weight change of the test animals between Day 1 and Day 6 was comparable to that observed in the corresponding control group animals over the same period.

Stimulation index was <3 at all three test concentrations (SI 0.92 at 2.5%, SI 1.29 at 5% and SI 1.77 at 10%)

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The substance has been tested and found not to be a skin sensitzer with a Stimulation index of <3.

In accordance with the classifciation criteria described in CLP Regulation 1272/2008 the substance is not classified for skin sensitization.

No information is available for respiratory sensitization however, the substance is a waxy solid with a low vapour pressure and therefore respiratory sensitization is considered unlikely.