2-Naphthalenol, 1-[2-[2-ethyl-4-[2-(2-ethylphenyl)diazenyl]phenyl]diazenyl]-, ar-heptyl derivs.

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

1

Reference substance name:

Reaction products of 2-Ethylaniline with heptyl naphthalen-2-ol

Cas Number:

1401000-07-4

Molecular formula:

Not applicable (UVCB Substance)

IUPAC Name:

Reaction products of 2-Ethylaniline with heptyl naphthalen-2-ol

Test material form:

solid

Specific details on test material used for the study:

Batch No.: 80-47-16
Storage Conditions: Room temperature, protected from light
Expiry Date: August 2017

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species/strain: WISTAR rats Crl: WI(Han)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female (non-pregnant and nulliparous)
Number of animals: 3 per step
Age at the
beginning of the study: 8-9 weeks


Housing and Feeding Conditions:

- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

In order to get the test item in a suspension, which is applicable to the animals, Polyethylenglycol (B. Merck, lot no. S7263585646, expiry date: 30 June 2017) was evaluated as vehicle and was considered to be adequate. This vehicle was chosen due to its non-toxic characteristics.
The test item was weighed out into a tared plastic vial on a precision balance.
Homogeneity of the test item in the vehicle was maintained by vortexing the prepared suspension thoroughly before each dose administration.
For all animals 2 g of the test item was suspended with the vehicle to gain a final volume of 10 mL and to achieve a dose of 2000 mg/kg body weight at a dose volume of 10 mL/kg body weight.
The dose formulations were made shortly before each dosing occasion.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

STEP 1: 3 female rats/2000 mg/kg body weight
STEP 2: 3 female rats/2000 mg/kg body weight

Control animals:

no

Details on study design:

Observation Period: All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.

Weight Assessment: The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

Clinical Examination: A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Mortality:

The test item showed no mortality but acute oral toxicity characteristics after a single dose administration.

Clinical signs:

other: The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, piloerection, hunched posture, sunken flunks, half eyelid-closure, prone position, discoloured faeces and dehydrati

Gross pathology:

No specific gross pathological changes were recorded for any animal.

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the conditions of the present study, a single oral application of the test item RED 2735 to rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity but no mortality.
The median lethal dose of RED 2735 after a single oral administration to female rats, observed over a period of 14 days is: LD50 cut-off (rat): 5000 mg/ kg bw.

According to Annex I of Regulation (EC) 1272/2008 the test item RED 2735 has no obligatory labelling requirement for toxicity and is not classified.
According to GHS (Globally Harmonized Classification System) the test item RED 2735 has obligatory labelling requirement for toxicity and is classified into Category 5.

Executive summary:

Two groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was suspended with / the vehicle Polyethylenglycol (PEG) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15.All animals were necropsied and examined macroscopically.

Table1:  Resultsper Step

Step		Sex / No.		Starting Dose (mg/kg bw)		Number of Animals		Number of Intercurrent Deaths
1		Female / 1 - 3		2000		3		0
2		Female / 4 - 6		2000		3		0

bw = body weight

All animals survived until the end of the study showing signs of toxicity.

The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, piloerection, hunched posture, sunken flunks, half eyelid-closure, prone position, discoloured faeces and dehydration. All symptoms recovered within up to 3 days post-dose.

Throughout the 14-day observation period, the weight gain of the animals was within the normal range of variation for this strain.

Macroscopic findings of surviving animals:

At necropsy, no macroscopic findings were observed in any animal of any step.