Reaction mass of rel-{(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl}methanol and rel-{(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl}methanol

Endpoint summary

Description of key information

An assessment of Contact Hypersensitivity to the registraton substance in the Mouse (Local Lymph Node Assay) was performed.

The data showed a dose-response and an EC3 value (the estimated test substance concentration that will give a SI =3) of 62.5% was calculated.

Mean DPM/animal values for the experimental groups treated with test substance concentrations 25, 50 and 100% were 922, 1129 and 1647 DPM, respectively. The mean DPM/animal value for the vehicle control group was 423 DPM. The SI values calculated for the substance concentrations 25, 50 and 100% were 2.2, 2.7 and 3.9, respectively. These results indicate that the test substance could elicit a SI ≥ 3.

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:

2015

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Specific details on test material used for the study:

GR-50-1408 is the Givaudan identification code which was employed for ROSYFOLIA during the early, developmental and testing period.

Species:

mouse

Strain:

other: CBA/J strain

Sex:

female

Details on test animals and environmental conditions:

Compared to sensitization tests using Guinea pigs, the Local Lymph Node Assay (LLNA) provides certain advantages with regard to animal welfare and scientific aspects.

Species Mouse, CBA/J strain, inbred, SPF-Quality. Recognized by the international guidelines as the recommended test system (e.g. OECD, EC, EPA).
Source: Janvier, Le Genest-Saint-Isle, France
Number of animals 20 females (nulliparous and non-pregnant), five females per group (main study only).
Age and body weight Young adult animals (approx. 10 weeks old) were selected. Body weight variation was within +/- 20% of the sex mean.
Identification Tail mark with marker pen.
Health inspection At least prior to dosing. It was ensured that the animals were healthy and that the ears were intact and free from any abnormality.


Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.
Animals were group housed in labeled Makrolon cages (MIII type; height 18 cm) containing sterilised sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany). Paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom) and shelters (disposable paper corner home, MCORN 404, Datesand Ltd, USA) were supplied as cage-enrichment. The acclimatization period was at least 5 days before the start of treatment under laboratory conditions. On Day 6, the animals were group housed in Makrolon MII type cages with a sheet of paper instead of sawdust and cage enrichment.
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
Free access to tap water.
Diet, water, bedding and cage enrichment evaluations for contaminants and/or nutrients were performed according to facility standard procedures. There were no findings that could interfere with the study.

Vehicle:

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

Concentration:

0 (Acetone/Olive oil (4:1 v/v)), 25, 50 and 100 % w/w

No. of animals per dose:

Three groups of five animals were treated with one test substance concentration per group.

Details on study design:

In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 25, 50 or 100% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Acetone/Olive oil (4:1 v/v)).
Three days after the last exposure, all animals were injected with 3 H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and pooled for each animal.
After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of Disintegrations Per Minute (DPM) and a stimulation index (SI) was subsequently calculated for each group.

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Mean DPM/animal values for the experimental groups treated with test substance concentrations 25, 50 and 100% were 922, 1129 and 1647 DPM, respectively. The mean DPM/animal value for the vehicle control group was 423 DPM

Parameter:

SI

Remarks on result:

other: The SI value calculated for the substance concentration 25 was 2.2 respectively.

Parameter:

SI

Remarks on result:

other: The SI value calculated for the substance concentration 50 was 2.7 respectively.

Parameter:

SI

Remarks on result:

other: The SI value calculated for the substance concentration 100 was 3.9 respectively.

Pre-screen test

No irritation and no signs of systemic toxicity were observed in any of the animals examined. Variations in ear thickness during the observation period were less than 25% from Day 1 pre-dose values. Based on these results, the highest test substance concentration selected for the main study was a 100% concentration.

Msin test

The slight irritation of the ear(s) as shown by the animals treated at 100% between Days 2 and 4 was considered not to have a toxicologically significant effect on the activity of the nodes.

No mortality occurred and no clinical signs 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.

The majority of auricular lymph nodes were considered normal in size, except for the nodes in the animals treated at 100% which were considered enlarged. No macroscopic abnormalities of the surrounding area were noted for any of the animals.

Mean DPM/animal values for the experimental groups treated with test substance concentrations 25, 50 and 100% were 922, 1129 and 1647 DPM, respectively. The mean DPM/animal value for the vehicle control group was 423 DPM. The SI values calculated for the substance concentrations 25, 50 and 100% were 2.2, 2.7 and 3.9, respectively.

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The results indicate that the test substance could elicit a SI ≥ 3. The data showed a dose-response and an EC3 value (the estimated test substance concentration that will give a SI =3) of 62.5% was calculated.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)