Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
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
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
Adopted: 17th December 2001
Deviations:
no
Remarks:
There were no significant deviations from the Good Laboratory Practice Regulations
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material: 4,4'-bis(2-methoxystyryl)-1,1'-biphenyl
- IUPAC name: 1-[(E)-2-(2-methoxyphenyl)ethenyl]-4-{4-[(E)-2-(2-methoxyphenyl)ethenyl]phenyl}benzene
- Molecular formula: C30H26O2
- Molecular weight: 418.53 g/mol
- Smiles notation: COC1=CC=CC=C1/C=C/C2=CC=C(C3=CC=C(/C=C/C4=CC=CC=C4OC)C=C3)C=C2
- InChl: 1S/C30H26O2/c1-31-29-9-5-3-7-27(29)21-15-23-11-17-25(18-12-23)26-19-13-24(14-20-26)16-22-28-8-4-6-10-30(28)32-2/h3-22H,1-2H3/b21-15+,22-16+
- Substance type: Organic
- Physical state: Solid

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Source: Velaz, Czech Republic
Sex:
female
Details on test animals and environmental conditions:
- Age at First Dose: 8-12 weeks; female animals were non-pregnant and nulliparous.

- Animal Health: Health condition of animals was examined by a veterinarian before initiation of the study.

- Acclimation: The animals were acclimated under the conditions identical to the conditions during the experiment 5 days prior to the start of treatment. The acclimation was according to the standard operation procedure.

- Housing Condition: The animals were housed in plastic cages suspended on stainless steel racks, up to 3 animals per cage in a room equipped with central air-conditioning. The average room temperature was maintained within the range of 23.2 ± 0.2 °C, relative humidity within 54.7 ± 2.3 %. The light regimen was set to a 12-hour light /12-hour dark cycle. Sanitation was performed according to the standard operation procedures.

- Diet: The laboratory food ssniff (Spezialdiäten GmbH, Germany) was offered at recommended doses each day approximately at the same time. The certificate of analysis is included in the raw data.

- Water: The animals received tap water for human consumption. Drinking water was supplied ad libitum. The quality of drinking water is periodical analysed and recorded; certificate of analysis is included in the raw data.

- Bedding: Lignocel S3/4, Lufa - ITL GmbH, Germany

- Animals Identification: The animals in the cage were marked by a line (I-III) on the tail with a waterproof marker. Each cage was marked with the study code, ID of animals and date of administration of the test item.

- Justification for the Choice of Species: Normally females are used for testing OECD TG 423 because females are typically the more sensitive gender.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
The test item was administered in a single dose by gavage using a metal stomach tube (administration volume 10 mL/kg. ( Oral administration simulates a potential route of human exposure, and oral dosing is recommended by regulatory agencies.)). Animals were fasted prior to dosing (food but not water were withheld overnight).
Doses:
5, 50, 300, and 2000 mg/kg body weight
No. of animals per sex per dose:
The starting dose could be selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg body weight. Available information indicated that the test item is likely to be non-toxic with regard to acute toxicity. A limit dose of 2000 mg/kg body weight was therefore used as a starting dose. One group of 3 females was dosed. Test item-related mortality was not observed during 24 hours and therefore, in a second step, another 3 females were treated at the same dose level.
Control animals:
not specified
Details on study design:
Animals were observed individually immediately after administration of the test item and 0.5, 1, 2, and 4 hours later. Each animal was inspected daily for the next 14 days.

All test animals were subjected to gross necropsy and the results were recorded for each animal whenever they died, survivors at the end of the observation period.

Individual weights of animals were measured immediately prior to test item administration and weekly thereafter. Weight differences after first and second weeks after administration were calculated and recorded.
Statistics:
No statistics were performed.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 - < 5 000 mg/kg bw
Based on:
test mat.
Mortality:
Five animals (5/6 females) survived the limit dose of 2000 mg/kg body weight.
Clinical signs:
Animals were observed individually immediately after administration of the test item and 0.5, 1, 2, and 4 hours later. Each animal was inspected daily for the next 14 days.
One animal was observed lethargy during 24 hour after administration and died on Day 2 post-treatment.
Five animal were observed no signs of intoxication, change of health, nor any other adverse reactions during 24 hours or 14-days observation period.
Body weight:
The body weight of animals increased during the study.
Gross pathology:
All animals were necropsied. During necropsy, no macroscopic findings were observed.

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The LD50 of the test item 4,4´-bis(2-methoxystyryl)-1,1´-biphenyl is greater than 2000 mg/kg and lower than 5000 mg/kg body weight after single oral administration to Wistar rats.
Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423 it can be concluded that the test item 4,4´-bis(2-methoxystyryl)-1,1´-biphenylis classified in GHS Category 5 (> 2000 – <5000) with a LD50 cut off value equal to 2500 mg/kg body weight, after single oral administration to Wistar rats.
Executive summary:

The purpose of the study was to evaluate the potential toxic effect of the test item 4,4´-bis(2-methoxystyryl)-1,1´-biphenyl when administered as a single oral dose to Wistar rats.

The procedure according to OECD Guideline 423 Acute Toxic Class(ATC)method was used. Available information indicated that the test item is likely to be non-toxic; therefore,a limit dose of 2000 mg/kg body weight was used as a starting dose.One group of 3 females was dosed. Test item-related mortality was not observed during 24 hours and therefore in a next step, 3 females were treated with the same dose.

The test item 4,4´-bis(2-methoxystyryl)-1,1´-biphenylwas administeredto 6 females Wistar rats at a limit dose of 2000 mg/kg.

Five females survived the limit dose. One female died on Day 2 post-treatment.During post-treatment time, the rest animals displayed neither signs of intoxication, change of health, nor any other adverse reaction.The body weight of animals increased during the study.

The macroscopical examination of the animals at the end of the study did not reveal treatment-related changes.

The LD50of the test item4,4´-bis(2-methoxystyryl)-1,1´-biphenylis greater than 2000 mg/kg and lower than 5000 mg/kg body weight after single oral administration to Wistar rats.

Based onAnnex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423itcan be concluded that thetest item 4,4´-bis(2-methoxystyryl)-1,1´-biphenylisclassified in GHS Category 5 (> 2000 – <5000) with a LD50cutoff value equal to 2500 mg/kg body weight,after single oral administration to Wistar rats.