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: oral

Administrative data

Endpoint:

acute toxicity: oral

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 420 (Acute Oral Toxicity - Fixed Dose Method)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

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. Adjustment was made for specific gravity of the test substance.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species: Rat, Wistar strain, Crl:WI (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 Pilot Study: 1 female. Main Study: 4 females per dose group (females were nulliparous and non-pregnant).
Age and body weight: Young adult animals (approx. 8-9 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification: Earmark
Health inspection: At least prior to dosing. It is ensured that the animals were healthy and without any abnormality that might affect the study integrity

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
Group housing of 3 animals per cage in labeled Makrolon cages (MIV 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.
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

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Remarks:

The test substance was dosed undiluted as delivered by the sponsor

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2.265 mL/kg body weight

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Based on resuts from the Pilot Study

Doses:

Single dose 2000 mg/kg (2.265 mL/kg) body weight.

No. of animals per sex per dose:

Four females were dosed at 2000 mg/kg body weight.

Control animals:

no

Details on study design:

Based on this "Pilot Study" results, the fixed dose level was selected for the main study where 4 females were dosed at 2000 mg/kg body weight via oral gavage and observed over a 14 day period. The animals were fasted prior to dosing and received food 3-4 hrs post-dosing. The animals were observed for signs or mortatlity twice daily, body weights recorded on days 1, 8 and 15. Clinical signs were observed on the day of dosing (Day1) and once daily until Day 15 and macroscopic examination performed after terminal necropsy.

Statistics:

None

Results and discussion

Preliminary study:

A "Pilot Study" was conducted in which females were orally administered the test substance at 2000 mg/kg body weight.. The test system and procedures were identical to those used during the main study. No mortality occurred. Hunched posture, piloerection and ptosis were noted on Days 1 and/or 2. Body weight gain was considered to be normal. No necropsy findings were noted. A fixed dose of 2000 mg/kg body weight was selected for the main study.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

other: Hunched posture and/or piloerection were noted for all animals between Days 1 and 7. Additionally, one animal showed flat posture, laboured respiration, lean appearance and ptosis on Days 1 and/or 2.

Gross pathology:

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

Applicant's summary and conclusion

Interpretation of results:

not classified

Conclusions:

The minimum oral lethal dose of Rosyfolia in rats was established to exceed 2000 mg/kg body weight. The LD50 cut-off value was considered to exceed 2000 mg/kg body weight.

Executive summary:

An acute oral toxicity study was performed in rats in accordance with OECD 420 guideline for "Acute Oral Toxicity-Fixed Dose Procedure" and EC, No440/2008, B1: Acute oral toxicity, Fixed Dose Procedure. Initially, in a Pilot Study, Rosyfolia was administered by oral gavage to one female Wistar rat at 2000 mg/kg body weight and no mortality occurred or signs of significant toxicity were observed. In this main study, four female fasted rats were administered a single dose of the test article via gavage at a dose of 2000 mg/kg body weight and observed for 14 days. Clinical signs were confined to hunched posture and/or piloerection, noted for all animals between Days 1 and 7. Additionally, one animal showed flat posture, labored respiration, lean appearance and ptosis on Days 1 and/or 2. There were no effects on body weight gain and by the end of the observation period, no deaths occurred. No macroscopic abnormalities were recorded. In this study, the acute oral LD₅₀for Rosyfolia was determined to be greater than 2000 mg/kg body weight.