Fatty acids, tall-oil, compds. with (Z)-N-9-octadecenyl-1,3-propanediamine (2:1)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 September 2017 to 20 October 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

May 2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Version / remarks:

March 2003

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF)

Version / remarks:

November 2000, including the most recent revisions

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

See attached justification. In short, based on the current data-set, it cannot be concluded if the outcome of the LLNA performed with Additiv 309 is a true or a false positive response. This implies that no conclusion can be drawn on classification for skin sensitizing properties. It is noted that classification for skin sensitizing properties at this point can be considered over-classification. Therefore further data are needed for a final conclusion on this endpoint. Since Additiv 309 is a UVCB with estimated low water solubility and high partition coefficient, the available in vitro/in chemico tests to address skin sensitizing properties are either technically not feasible to perform or are expected to yield a result that might not be reliable (false negative outcome). In order to allow final conclusion on skin sensitizing properties of Additiv 309, it was therefore concluded that performance of a GPMT study was necessary.

Test material

Test material information

Constituent 1

Reference substance name:

Additiv 309

IUPAC Name:

Additiv 309

Test material form:

liquid: viscous

Details on test material:

Sponsor's Identification: Optimol Additiv 309
Appearance: Pasty dark brown liquid
Batch: 0000769695
Expiry date: 27 June 2021
Storage conditions: room temperature in the dark

Specific details on test material used for the study:

Specific gravity / density: <1g/cm3 at 20 degrees C

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle, France
- Females nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: SPF
- Age at study initiation: approx. 4 weeks
- Weight at study initiation: 301-353 g
- Housing: Group housing of maximally 5 animals per labeled Noryl cage containing sterilized sawdust as bedding material and shelters as cage enrichment.
- Diet: Complete maintenance diet for guinea pigs (MS-H, SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum. In addition, hay was provided at least twice a week.
- Water: free access to tap water
- Acclimation period: at least 5 days
- Indication of any skin lesions: Before experimental start it was ensured that the animals were healthy and that the skin to be treated was intact and free from any abnormality.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-22
- Humidity (%): 46 - 65
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

-IN-LIFE DATES: From: 20 September 2017 To: 20 October 2017

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

corn oil

Concentration / amount:

1%

Day(s)/duration:

Day1

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Induction 2

Route:

other: provocation mild inflammatory reaction

Vehicle:

water

Concentration / amount:

10% SDS

Day(s)/duration:

Day 7, 24 hours before epidermal induction,

Induction 3

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Concentration / amount:

100%

Day(s)/duration:

Day 8, 48 hours

Adequacy of induction:

other: 100% was well-tolerated systemically and did not cause skin irritation. Hence SDS was used to provoke a mild inflammatory reaction.

Challenge

No.:

#1

Route:

epicutaneous, semiocclusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

100%

Day(s)/duration:

Day 21, 24 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10 (experimental group), 5 (control group)

Details on study design:

RANGE FINDING TEST:
A preliminary irritation study was conducted in order to select test item concentrations to be used in the main study. The starting- and subsequent concentrations were taken from the series: 100% (undiluted), 50%, 20%, 10%, 5%, 2%, 1%. The test system and procedures we re identical to those used during the main study. The four animals selected were between 4 and 9 we eks old. No body weights were determined.

Intradermal injections: A series of four test item concentrations was tested, the highest concentratio n being the maximum concentration that could technically be injected. Two animals received two dif ferent concentrations in duplicate (0.1 mL/site) in the clipped scapular region. The injection sites were assessed for irritation 24 and 48 hours after treatment.

Epidermal application: A series of four test item concentrations was tested, the highest concentration being the maximum concentration that could technically be applied. Two different concentrations were applied (0.5 mL each) per animal to the clipped flank, using Metalline patches (2x3 cm) mounted on Medical tape which were held in place with Micropore tape and subsequently Coban elastic bandage. The animals receiving intradermal injections were treated with the lowest concentrations and two other animals with the highest concentrations.

After 24 hours, the dressing was removed and the skin cleaned of residual test item using water. The treated skin areas were assessed for irritation 24 and 48 hours after removal of the dressings.

MAIN STUDY

A. INDUCTION EXPOSURE
- No. of exposures: 2 (injection day 1, epidermal exposure day 8)
- Dermal observations: The dermal reactions caused by the intradermal injections were assessed for irritation on day 3
- Groups: One test substance group and one control group
- Site: Scapular region (clipped)
- Concentrations: 1% for intradermal, 100% for epicutaneous

B. CHALLENGE EXPOSURE
- No. of exposures: 1 (day 21)
- Exposure period: 24 hours (after removal of the dressing, the skin was cleaned of residual test item and vehicle using water)
- Groups: One test substance group and one control group
- Site: flank (clipped)
- Concentrations: 100%
- Evaluation (hr after challenge): 24 and 48 hours after removal of the dressing

Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were weighed individually on day 1 (predose) and at termination of the study.

Challenge controls:

Results of a reliability study are included.

Positive control substance(s):

yes

Remarks:

Results of a reliability check performed with alpha-hexylcinnemaldehyde are included in the report.

Results and discussion

Positive control results:

Reliability check performed in July 2017: all experimental animals gave a positive response to the 50% test item concentration in the challenge phase at 24 hours after the challenge exposure (8/10: score 2; 2/10: score 1). This was considered indicative of sensitisation, based on the absence of any response in the control animals. These results lead to a sensitisation rate of 100 percent to the 50% concentration.

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

1% intradermal, 100% epidermal, 100% challenge

No. with + reactions:

8

Total no. in group:

10

Remarks on result:

positive indication of skin sensitisation

Results 2

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

no indication of skin sensitisation

Any other information on results incl. tables

At 24 hours after challenge, 8 out of 10 test animals showed discrete or patch erythema (grade 1). After 48 hours, discrete or patch erythema were still visible in 3/10 guinea pigs in the test substance group. Scaliness was observed in 7/10 animals in the test substance group (all of these showed positive signs of skin sensitization).The reactions noted in the experimental animals after the epidermal induction exposure were considered to be enhanced by the SDS treatment. No skin reactions were evident in the control animals.

No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

Applicant's summary and conclusion

Interpretation of results:

Category 1A (indication of significant skin sensitising potential) based on GHS criteria

Conclusions:

Based on a Guinea Pig Maximization Test conducted according to OECD/EC guidelines and GLP principles, Optimol Additiv 309 is considered to have skin sensitising properties (Cat. 1A according to Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) of substances and mixtures).

Executive summary:

A Guinea Pig Maximization Test was conducted according to OECD/EC guidelines and GLP principles. Ten experimental animals were intradermally injected with a 1% concentration and epidermally exposed to a 100% concentration. Five control animals were similarly treated, but with vehicle alone (corn oil). Doses were established based on the outcome of a preliminary irritation study. Approximately 24

hours before the epidermal induction exposure all animals were treated with 10% SDS. Two weeks after the epidermal application all animals were epidermally challenged with a 100% test item concentration and the vehicle. No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. Discrete or patchy erythema in 80% (8 out of 10) of the animals was observed at 24 hours after challenge which was still present in 33% (3 out of 10) of the animals at 48 hours after exposure. Scaliness was seen on some treated skin sites. No skin effects were seen in the controls. Based on the positive response in 80% of the animals at 24 hours after exposure, Optimol Additiv 309 is considered to have skin sensitising properties and is classified Cat. 1A according to Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) of substances and mixtures.