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

Oral: measured LD50 > 2000 mg/kg bw, and the estimated LD50 cut-off can be considered to be > 2000 mg/kg bw, female rat, OECD TG 420, 2015

Dermal: measured LD50 > 2000 mg/kg bw for males and for the sexes combined and within 1000 -2000 mg/kg bw for female rats, OECD TG 402, 2015

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

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.

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

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.

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.

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

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.

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.

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The available information as a whole meets the tonnage driven information requirements of REACH.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

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.

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

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

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.

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

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.

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The available information as a whole meets the tonnage driven information requirements of REACH

Additional information

ORAL

Key study : OECD TG 420, 2015 :

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 tohunched 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. Applicant assessment indicates: under the conditions of this study the acute toxicity estimate would be considered to be > 5000 mg/kg bw.

DERMAL:

Key study : OECD TG 402, 2015 : The study was performed according to OECD TG 402 and EU Method B.3 Acute Toxicity (Dermal) and in accordance with GLP.

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. Applicant assessment indicates: under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight on the basis of absence of significant clinical toxicological effects and/or bodyweight increases in all males/females.

Justification for classification or non-classification

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: oral

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: dermal