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Diss Factsheets

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
6 July 2021 - 10 August 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2021
Report date:
2021

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
December 2001
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
December 2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EC No 440/2008 Part B. Acute Oral Toxicity, Acute Toxic Class Method
Version / remarks:
May 2008
Deviations:
no
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 200, including the most recent revisions
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
(3aS,3bS,9aR,9bS,11aS)‐11a‐ethyl‐10‐methylidene‐1H,2H,3H,3aH,3bH,4H,5H,7H,8H,9H,9aH,9bH,10H,11H,11aH‐cyclopenta[a]phenanthrene‐1,7‐dione
Cas Number:
54024-17-8
Molecular formula:
C20 H26 O2
IUPAC Name:
(3aS,3bS,9aR,9bS,11aS)‐11a‐ethyl‐10‐methylidene‐1H,2H,3H,3aH,3bH,4H,5H,7H,8H,9H,9aH,9bH,10H,11H,11aH‐cyclopenta[a]phenanthrene‐1,7‐dione
Test material form:
solid: particulate/powder
Details on test material:
Storage Conditions: In refrigerator (2-8°C)
Specific details on test material used for the study:
Physical description: Off-white powder
Test item handling: No specific handling conditions required
CAS number: 54024-17-8
Molecular formula: C20H26O2
Molecular weight: 298.42
Purity correction factor: No correction factor required

Test animals

Species:
rat
Strain:
other: Crl: WI(Han)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat: Not applicable
- Source: Charles River France, L’Arbresle, France
- Females (if applicable) nulliparous and non-pregnant: yes
- Rationale for use of males: Not applicable
- Age at study initiation: Young adult animals (approximately 8-11 weeks old)
- Weight at study initiation: 147 to 197 g
- Fasting period before study: Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the test item. Water was available.
- Housing: On arrival and following assignment to the study, animals were group housed (up to 3 animals of the same sex and same dosing group together) in polycarbonate cages containing sterilized wooden fibers as bedding material equipped with water bottles. Animals were separated during designated procedures/activities
- Historical data: Not reported
- Diet: ad libitum; Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water: ad libitum; Municipal tap-water
- Contaminants: It is considered that there were no known contaminants in the feed and water that would interfere with the objectives of the study.
- Acclimation period: at least 5 days
- Microbiological status when known: SPF
- Method of randomisation in assigning animals to test and control groups: Animals were assigned to the study, with all animals within ± 20% of the sex mean body weights. Animals in poor health or at extremes of body weight range were not assigned to the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 22 (actual temperature); 18 to 24 (target temperature)
- Humidity (%): 53-73 (actual humidity); 40-70 (target humidity); Values outside target range were without a noticeable effect on the clinical condition of the animals or on the outcome of the study
- Air changes (per hr): Ten or greater air changes per hour with 100% fresh air
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 7 July 2021 To: 10 August 2021

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: Not reported
- Amount of vehicle: Not reported
- Justification for choice of vehicle: Trial preparations were performed to select the suitable vehicle and to establish a suitable formulation procedure. These trials were not performed as part of this study and these preparations were not used for dosing
- Lot/batch no.: Not reported
- Purity: 1.125 (specific gravity)

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: The test item was stored in the refrigerator and was allowed to warm to room temperature for at least 30 minutes before use. Test item dosing formulations (w/w) were homogenized to visually acceptable levels at appropriate concentrations to meet dose level requirements. The dosing formulations were kept at room temperature until dosing. The dosing formulations were stirred until and during dosing.
An adjustment was made for specific gravity of the vehicle. No correction was made for the purity/composition of the test item.

CLASS METHOD
- Rationale for the selection of the starting dose: The dose levels were based on the OECD test guidelines and were selected from the series 5 (lowest dose level), 50, 300 and 2000 (highest dose level) mg/kg body weight. The starting dose level should be the one that is likely to produce mortality in at least some of the animals and was selected based on available toxicity data of the test item.
- The toxicity of the test item was assessed by stepwise treatment of groups of 3 females. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups. The first group was treated at a dose level of 300 mg/kg. Based on the results, two additional groups were dosed at 2000 mg/kg.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
300 mg/kg bw: three females
2000 mg/kg bw: two groups of three females (six in total)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: at periodic intervals on the day of dosing (at least three times) and once daily thereafter
- Frequency of weighing: Day 1 (pre-dose), 8 and 15
- Necropsy of survivors performed: yes
- Clinical signs: All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate), particular attention being paid to the animals for the first hour after dosing.
- Other examinations performed: mortality/morbundity checks, twice daily
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occured.
Clinical signs:
other: At 300 mg/kg, hunched posture was noted in all animals and erected fur in two animals between Days 1 and 3. At 2000 mg/kg, hunched posture was noted in all animals on Day 1 and/or 2. In addition, three animals showed uncoordinated movements on Day 1 and t
Gross pathology:
No test item related abnormalities were found at macroscopic postmortem examination of the animals.
One incidental finding included reddish foci on the thymus in one animal, which is more often seen in rats of this strain in this kind of studies and is therefore not toxicological relevant.

Applicant's summary and conclusion

Interpretation of results:
other: Not classified according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments)
Conclusions:
An acute oral toxicity study was performed in Wistar Han rats according to OECD 423 and in accordance with GLP principles. The oral LD50 value of the test substance was established to exceed 2000 mg/kg bw. Based on the results, the test substance does not have to be classified according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).
Executive summary:

An acute oral toxicity study was performed in Wistar Han rats according to OECD 423 and in accordance with GLP principles. The objective of this study was to assess the toxicity of the test item when administered in a single dose to female rats at one or more defined dosages. Initially, EMETAM was administered by oral gavage to three female Wistar Han rats at 300 mg/kg body weight. In a stepwise procedure two additional groups of three females were dosed at 2000 mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).
No mortality occurred.
At 300 mg/kg, hunched posture was noted in all animals and erected fur in two animals between Days 1 and 3. At 2000 mg/kg, hunched posture was noted in all animals on Day 1 and/or 2. In addition, three animals showed uncoordinated movements on Day 1 and three animals erected fur on Days 1 and 2.
The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
No test item related abnormalities were found at macroscopic postmortem examination of the animals.
The oral LD50 value of EMETAM in Wistar Han rats was established to exceed 2000 mg/kg body weight. Based on the results, the test substance does not have to be classified according to Regulation (EC) No 1272/2008 on classification, labelling and packagin of items and mixtures (including all amendments).