Sebacohydrazide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 June - 17 July 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF),Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau. November 2000, including the most recent revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sebacohydrazide

EC Number:

213-126-8

EC Name:

Sebacohydrazide

Cas Number:

925-83-7

Molecular formula:

C10H22N4O2

IUPAC Name:

sebacohydrazide

Test material form:

solid: particulate/powder

Details on test material:

Test item name (as stated in report): SDH
Test item storage: at room temperature protected from light

Test animals

Species:

rat

Strain:

other: Crl:WI (Han)

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 8-10 weeks old).
- Weight at study initiation: 139 to 178 g. Body weight variation was within +/- 20% of the sex mean.
- Fasting period before study: overnight (maximum 20 hours)
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 – 22
- Humidity (%): 32 - 72
- Air changes (per hr): approx 10
- Photoperiod (hrs dark / hrs light): 12/12

Deviations from the maximum (1 day) or minimum (2 days) level of daily mean target humidity occurred. This study plan deviation is considered not to have affected the integrity of the study because it did not noticeably affect the clinical condition of the animals or the outcome of the study.

IN-LIFE DATES: From: 05 June to 17 July

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle:Trial preparations were performed at the Test Facility to select the suitable vehicle and to establish a suitable formulation procedure. Trial preparation formulations were not used for dosing and were discarded after the assessment is complete. These trial preparations have a non-GLP status and were carried out in the quality assured environment of the Test Facility.

DOSE VOLUME APPLIED: 10 mL/kg body weight


DOSAGE PREPARATION: 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. No adjustment was made for specific gravity of the vehicle. No correction was made for the purity/composition of the test item. Any residual volumes were discarded.


CLASS METHOD
The first group was treated at a dose level of 2000 mg/kg. Based on the results, an additional group was dosed at 2000 mg/kg after which a group had to be dosed at 300 mg/kg, which was inadvertently dosed at 50 mg/kg. Subsequently, two additional groups were dosed at 300 mg/kg to comply with the dosing regimen as described in the guidelines.

Doses:

2000 mg/kg body weight
300 mg/kg body weight

No. of animals per sex per dose:

2000 mg/kg: 6 (2 groups of three females in a stepwise manner)
300 mg/kg: 6 (2 groups of three females in a stepwise manner)

Control animals:

no

Details on study design:

Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.

- Duration of observation period following administration: 14 days
- Mortality/Viability: Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day.Terminal body weight were collected from animals found dead or euthanised moribund after day 1.
- Body weights: Animals were weighed individually on day 1 (predose), 8 and 15. A fasted weight was recorded on the day of dosing.
- Clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15.
- Necropsy of survivors performed: All moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure at the end of the observation period. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.
- Other examinations performed: none.

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:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, no mortality occurred for the first dose group and all three animals of the second dose group were killed in extremis on Day 8.
At 300 mg/kg, no mortality occurred.
At 50 mg/kg, no mortality occurred.

Clinical signs:

At 2000 mg/kg, for the animals killed in extremis, hunched posture, uncoordinated movements, piloerection, lean appearance and yellow urine were noted up to the day of sacrifice.
At 2000 mg/kg, for the surviving animals, hunched posture and piloerection were noted for all animals on day 1 and hunched posture and lean appearance were noted between days 8 and 11 for one single animal.
At 300 mg/kg, hunched posture, uncoordinated movements and piloerection were noted between days 1 and 4.
At 50 mg/kg, hunched posture and piloerection were noted on day 1.

Body weight:

For the animals killed in extremis, severe body weight loss (>10% mean weight) was noted on day 8.
The mean body weight gain shown by the surviving animals over the study period was considered to be normal.

Gross pathology:

Accentuated lobular pattern and/or pale discoloration of the liver, reduced thymus size and general emaciation were found in the animals that were sacrificed for humane reasons during the study, at macroscopic post mortem examination. Macroscopic examination of the surviving animals at termination did not reveal any abnormalities.

Applicant's summary and conclusion

Interpretation of results:

other: Acute tox 4

Remarks:

according to EC No 1272/2008

Conclusions:

The oral LD50 value of SDH in Wistar rats was established to be within the range of 300- 2000 mg/kg body weight.

Executive summary:

An acute oral toxicity study with in rats was performed according to OECDC/EC guidelines and in accordance with GLP principles. SDH was administered by oral gavage to one group of three female rats at 2000 mg/kg body weight. In a stepwise procedure, four additional groups of three females were dosed at 2000 (1 group), 50 (1 group) and 300 (2 group) mg/kg body weight. All 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).

Two groups were administered with 2000 mg/kg. In the first group, no mortality occurred. Hunched posture and piloerection were noted for all animals on day 1 and hunched posture and lean appearance were noted between days 8 and 11 for one single animal. For the second group, severe body weight loss was noted on day 8, therefore all three animals were killed in extremis on day 8. Also hunched posture, uncoordinated movements, piloerection, lean appearance and yellow urine were noted up to the day of sacrifice. Macroscopic evaluation findings were found in liver (accentuated lobular pattern and/or pale discoloration) and thymus (reduced size). General emaciation was also found. Next, one group was inadvertently dosed at 50 mg/kg instead of 300 mg/kg. This deviation did not affect the integrity of the study because additional groups were added at the required dose levels. In this group no mortality occurred. Hunched posture and piloerection were noted on day 1. Lastly, two groups were administered with 300 mg/kg. No mortality occurred. Hunched posture, uncoordinated movements and piloerection were noted between days 1 and 4. The mean body weight gain shown by the surviving animals over the study period was considered to be normal. Macroscopic examination of the surviving animals at termination did not reveal any abnormalities.

The oral LD50 value of SDH in Wistar rats was established to be within the range of 300 -2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2000 mg/kg body weight. Based on these results, SDH should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.