N,N'-bis(2,4,6-triisopropylphenyl)methanediimine

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-04-30 - 2014-09-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

under GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

OECD - Guideline for Testing of Chemicals No. 423 -"Acute Oral Toxicity- Acute Toxic Class Method"; adopted: 17th December 2001

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

EEC Directive 440/2008 Part B -Method B.1.tris, test methods pursuant to Regulation (EC) No 1907/2006 (REACH), EU Directive 67/548/EEC and EC Regulation 1272/2008

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-bis[2,4,6-tris(propan-2-yl)phenyl]methanediimine

EC Number:

815-171-4

Cas Number:

300382-79-0

Molecular formula:

C31H46N2

IUPAC Name:

N,N'-bis[2,4,6-tris(propan-2-yl)phenyl]methanediimine

Test material form:

liquid

Remarks:

yellowish

Details on test material:

Storage: Room temperature

Test animals

Species:

rat

Strain:

Wistar

Remarks:

RCCHan: WIST

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan GmbH, 5960 AD Horst, Netherlands
- Females (if applicable) nulliparous and non-pregnant: Yes, at start of the study the animals were nulliparous and non-pregnant and free of all clinical symptoms or diseases.
- Age at study initiation: 8 - 12 weeks approximately
- Weight at study initiation: 153g - 176g
- Fasting period before study: Yes. For administration, food was withheld from the animals for approximately 16 - 24 h before administration of the test item, and they were fed again approximately 2 - 4 h after administration.
- Housing: The animals were group caged conventionally in polycarbonate cages on low dust wood granulate bedding ("SsniffBF 1, Brandenburg Holzfaserstoffe GmbH & Co. KG, Germany'''). The cages of the animals were placed on racks. The wood granulate was randomly checked for contaminants at regular intervals. The analyses yielded no evidence of any adverse effects on the aim of the study. Wooden blocks for environmental enrichment were added to each cage. As soon as necessary, they were replaced by new ones. The cages were changed at least once a week. Feed racks and water bottles were not changed. All cage material was washed with hot water. In the first stage of the washing programs an alkaline cleaning agent (Neodisher Alka 300; Chemische Fabrik Dr. Weigert GmbH & Co. KG, concentration: 2.2 g/L) was used.
The animal room was cleaned and disinfected weekly. A continuous pest control was performed using a cockroach trap without pesticides (e.g. Killgerm Roach Trap, Killgerm GmbH, 41460 Neuss, Germany). The contact ofthe animals with the traps was avoided in any case.
- Diet (e.g. ad libitum): standard diet "ssniffR/M-H 10 mm (Ratten-/MäuseHaltungsf.) V1534-0, ssniff Spezialdiäten GmbH, 59494 Soest", ad libitum, available from racks in the lid of the cage.
The nutritive composition and the contaminant content of the standard diet were checked and analyzed routinely in random samples. Nothing untoward was found.
- Water (e.g. ad libitum): tap water ad libitum from polycarbonate bottles
The tap water was of drinking water quality (according to the Drinking Water Decree in the current version).
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): 55 ± 5 %
- Air changes (per hr): approx. 10 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours rhythm

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): The administration volume was 10 mL/kg body weight.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg bw

DOSAGE PREPARATION (if unusual):
The test item was formulated in com oil. The applied formulations were well mixed before administration. The formulations for administration were prepared at room temperature.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
Three animals were used for each step. The dose level to be used as the starting dose was selected from one of four fixed levels, 5, 50, 300 and 2000 mg/kg body weight. The starting dose level should be that which is most likely to produce mortality in some of the dosed animals. Absence or presence of compound-related mortality of the animals do:,ed at one step determined the next step, i.e.:
• no further testing is needed,
• dosing of three additional animals, with the same dose,
• dosing of three additional animals at the next higher or the next lower dose level.
The test item was tested using a stepwise procedure, each step using three animals of a single sex. The procedure is described in the flow charts of Annex 2, OECD guideline 423. The starting dose selected was 2000 mg/kg bw.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

2 x 3 females

Control animals:

no

Remarks:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Animals were observed individually after dosing at least once during the first 30 minutes, periodically during the first 24 hours, with individual attention given during the first 4 hours, and twice daily (except weekend and public holidays) for at least 14 days thereafter.
Clinical signs and mortality rates were determined several times on the day of application and subsequently at least once daily for an observation period of at least 14 days. Mortality and in the event of symptoms occurring, nature and duration were recorded individually. The day of application is defined as day 1. Times after application until the following day were recorded either in minutes or in hours, depending on what was appropriate. The duration of the symptoms and the times of death are given relative to the time of application to the individual animal. The real time points can be taken from the raw data. In general, death was taken as a symptom. If no symptoms were seen until death, time of death was taken as the first occurrence of a symptom. In the results section, the findings are summarized.
The weight gain of the animals was checked weekly until the end of the study. The weight gain of the animals was calculated based on rounded individual values. The weights are given in grams (g). Indicated under the heading day 8 are e.g. the data obtained on the 7th day after administration.
- Necropsy of survivors performed:
Animals which died or were killed in moribund state were weighed (except on day
of administration) and dissected as soon as possible, and examined macroscopically.
The surviving animals were sacrificed by carbon dioxide at the end of the study, dissected and examined macroscopically.
- Other examinations performed: clinical signs, body weight

Statistics:

Collection, Processing and Evaluation of Data
During this study collection, storage and evaluation including statistics of in-life data was done on- or offline by using the validated Pristima System produced by Xybion Medical Systems Corporation, 240 Cedar Knolls Road, Cedar Knolls, New Jersey 07927, USA.
Data of the necropsies were collected offline and were entered by key entry.

Calculation of the LD50
The LD50 value was estimated according to OECD - Guideline for Testing of Chemicals No. 423 -"Acute Oral Toxicity- Acute Toxic Class Method"; adopted: December 17, 2001.

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Effect levels 2

Sex:

female

Dose descriptor:

LD0

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No deaths occurred.

Clinical signs:

other: No clinical signs were observed.

Gross pathology:

The necropsies performed at the end of the study revealed no particular findings.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

EU implementation

Conclusions:

The study was conducted under GLP according to OECD 423 on the registered substance itself. The method is to be considered scientifically reasonable with no deficiencies, hence, the results are considered to be sufficiently reliable to assess the acute oral toxicity of the test item in rats.
A dose of 2000 mg/kg body weight was tolerated by female rats without mortalities, clinical signs, effects on weight gain and gross pathological findings.
According to OECD guideline 423 the LD50 cut-off of Bis(2,4,6-triisopropylphenyl)carbodiimide is > 5000 mg/kg bw. According to EU Directive 67/548/EEC and EC Regulation 1272/2008 the test item is unclassified. So it is regarded as non-toxic after oral application.

Executive summary:

This study was performed according to OECD 423 under GLP to assess the acute oral toxicity of Bis(2,4,6-triisopropylphenyl)carbodiimide to Wistar rats. The test item was formulated in corn oil; the administration volume was 10 mL/kg body weight, it was administered to six female rats.

A dose of 2000 mg/kg body weight was tolerated by female rats without mortalities, clinical signs, effects on weight gain and gross pathological findings. According to OECD guideline 423 the LD50 cut-off of Bis(2,4,6-triisopropylphenyl)carbodiimide is > 5000 mg/kg bw. According to EU Directive 67/548/EEC and EC Regulation 1272/2008 the test item is unclassified. So it is regarded as non-toxic after oral application.