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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study performed under GLP. All relevant validity criteria were met.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

[(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1R,2S)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol

EC Number:

942-597-9

Cas Number:

1655500-83-6

Molecular formula:

C12H22O

IUPAC Name:

[(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1R,2S)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl]methanol; [(1S,2R)-1-methyl-2-[(2S)-5-methylhex-4-en-2-yl]cyclopropyl]methanol

Test material form:

other: liquid

Details on test material:

Batch VE00340479
Purity 87.8% (Sum of the four isomers)

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.
The test substance was dosed undiluted as delivered by the sponsor. The test substance was kept at room temperature protected from light for a maximum of 4 hours prior to dosing. Based on the test substance data provided by the sponsor, it was considered that the test substance remained stable during this relatively short time period.
No correction was made for the purity/composition of the test substance

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species: Rat, Wistar strain, Crl:WI (Han) (outbred, SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC).
Source: Charles River Deutschland, Sulzfeld, Germany.
Number of animals 5 males and 5 females at 2000 mg/kg; 5 females at 1000 mg/kg, (females were nulliparous and non-pregnant),
Age and body weight: Young adult animals (approx. 10 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification: Tail mark with indelible ink.
Health inspection At least prior to dosing. It was ensured that the animals were healthy and that the skin to be treated was 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.
Accommodation
Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom). Acclimatization period was at least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm).
Free access to tap water and to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Remarks:

The test substance was dosed undiluted as delivered by the sponsor.

Details on dermal exposure:

Rosyfolia was applied to the skin undiluted, single application. The test substance was applied on an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The test substance was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D)* ,successively covered with aluminum foil and Coban elastic bandage* . A piece of Micropore tape* was additionally used for fixation of the bandages in females only.

Duration of exposure:

24 hours, after which dressings were removed and the skin cleaned of residual test substance using tap water.

Doses:

2000 mg/kg (2.265 mL/kg) body weight
1000 mg/kg (1.133 mL/kg) body weight

No. of animals per sex per dose:

2000 mg/kg (2.265 mL/kg) body weight, 5 males and 5 females,
1000 mg/kg (1.133 mL/kg) body weight, 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality and viability recorded 2x/day; clinical signs were monitored at periodic intervals on the day of dosing (Day1) and once daily until Day 15; body weight measurement performed on Days 1, 8, 15 and at death if found dead after Day 1
- Necropsy of survivors performed: yes
- Clinical signs : The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1)

Statistics:

None

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

ca. 1 000 - ca. 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, one male and three females were found dead on Day 2.
At 1000 mg/kg, no mortality occurred.

Clinical signs:

other: At 2000 mg/kg, lethargy, abnormal posture, flat posture, hunched posture, uncoordinated movements, head drop, rales, shallow respiration, piloerection, chromodacryorrhoea, hypersensitivity to touch, ptosis and/or hypothermia were noted for the animals. By

Gross pathology:

No abnormalities were found at macroscopic post mortem examination in any of the animals.

Applicant's summary and conclusion

Interpretation of results:

harmful

Conclusions:

The dermal LD50 value of Rosyfolia in Wistar rats was established to exceed 2000 mg/kg body weight for males and for the sexes combined and within 1000-2000 mg/kg body weight for females.

Executive summary:

The oral toxicity of rosyfolia was assessed when rats were administered a single dermal application. Initially rosyfolia was administered (n=5/sex) at 2000 mg/kg body weight for 24hrs. Based on the results, one additional group of five females was dosed at 1000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15). At 2000 mg/kg, one male and three females were found dead on Day 2. At 1000 mg/kg, no mortality occurred. At 2000 mg/kg, lethargy, abnormal posture, flat posture, hunched posture, uncoordinated movements, head drop, rales, shallow respiration, piloerection, chromodacryorrhoea, hypersensitivity to touch, ptosis and/or hypothermia were noted for the animals. The surviving animals had recovered from the symptoms by Day 7. General erythema, scales and/or scabs were seen in the treated skin-area of the animals during the observation period. At 1000 mg/kg, no clinical signs of systemic toxicity were noted. General erythema, erythema maculate, scales were seen in the treated skin-area of the animals during the observation period. The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity. No abnormalities were found at macroscopic post mortem examination in any of the animals. The dermal LD50 value of rosyfolia in Wistar rats was established to exceed 2000 mg/kg body weight for males and for the sexes combined and within 1000-2000 mg/kg body weight for females.