Formamide, N-[(2-bromo-5-hydroxy-4-methoxyphenyl)methyl]-N-[2-(4-hydroxyphenyl)ethyl]-

Endpoint summary

Description of key information

Acute toxicity: Oral: In an acute oral toxicity study in female Crl:WI (Han) (SPF) rats, following the acute toxic class method in accordance with the OECD Guideline 423 and EU Method B.1 tris, the LD50 was established to exceed 2000 mg/kg. According to the OECD 423 guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight (Latour, 2016).

Acute toxicity: Inhalation: No reliable acute toxicity study via inhalation were available. However, this endpoint is waived as specific data are available for the oral exposure route.

Acute toxicity: Dermal: An acute dermal toxicity study does not have to be performed based on the following justification:

• The substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route: LD50>2000 mg/kg bw (OECD 423 study) and no clinical signs observed leading to STOT SE classification. In addition, no abnormalities were found at macroscopic post mortem examination of all animals.
• No systemic effects have been observed in in vivo studies with dermal exposure:
• Skin irritation: not irritating to the skin in an OECD 404 test
• Skin sensitization: the substance was not regarded as a skin sensitizer in a test comparable to the OECD 429 Guideline. No signs of systemic toxicity were noted for the animals in the pre-screen test and no mortality occurred and no clinical signs of systemic toxicity were observed in the animals of the main study.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-11-24 to 2016-12-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Test Material SOURCE OF TEST MATERIAL
- Batch No.of test material: I15EB1917
- Expiration date of the lot/batch: 13 June 2017 (retest date)
- Purity: 98.9%
- Purity correction factor: 1

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- Solubility and stability of the test substance in the solvent/vehicle: Analysis of test item in vehicle for concentration, stability, homogeneity was not performed, however preparation was performed with approved procedure and documented in detail. Homogeneity was visually inspected prior to use.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test preparation material was kept at room temperature no more than 4 hours before animals were dosed.

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han) (outbred, SPF-Quality)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: young adult animals: approximately 9 weeks
- Weight at study initiation: 161 - 175 grams
- Fasting period before study: animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test item. Water was available ad libitum.
- Housing: group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): ad libitum, free access to pelleted rodent diet
- Water (e.g. ad libitum): ad libitum, free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24 °C
- Humidity (%): 40-70%; study plan deviation: deviations from the minimum level of daily mean relative humidity occurred. Evaluation: laboratory historical data did not indicate an effect of the deviations. The study integrity was not affected by this deviation.
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): no data
- Justification for choice of vehicle: The vehicle was selected based on trial preparations performed at Charles River Den Bosch and on test item data supplied by the Sponsor. The vehilce was chosen from (in order of preference): water (Elix) (test item did not dissolve), 1% aq. carboxymethyl cellulose (test item did not dissolve), propylene glycol (spec. gravity 1.036)(suspension), polyethylene glycol 400 (spec. gravity 1.125) (not tested according to vehicle selection criteria) and corn oil (spec. gravity 0.92).

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION:
- The preparations (w/w) were kept at room temperature and were dosed within 4 hours after adding the vehicle to the test item. Homogeneity was obtained to visually acceptable levels and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies.
- Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test item as the correction factor is 1.

Doses:

2000 mg/kg (single dosage)

No. of animals per sex per dose:

3 females per dose group (2 dose groups in total)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (until day 15)
- Frequency of observations and weighing:
mortality/viability: twice daily
body weights: days 1 (pre-administration), 8 and 15; study plan deviation: bodyweights of the first three animals on Day 15 were not recorded correctly, due to a technical error. Evaluation: Sufficient data is available for evaluatio of the study. The study integrity was not affected by these deviations.
clinical signs: at periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded: maximum grade 4: grading slight (1) to very severe (4); maximum grade 3: grading slight (1) to severe (3); maximum grade 1: presence is scored (1).
- Necropsy of survivors performed: yes, at the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

No statistical analysis was performed.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred
Number of dead animals in each test group with 3 animals each:
Females 2000 mg/kg: 0
Females 2000 mg/kg: 0

Clinical signs:

other: Hunched posture, piloerection and uncoordinated movements were noted for all animals between Days 1 and 4.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of all animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 value of T002078 in Wistar rats was established to exceed 2000 mg/kg body weight. According to OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Based on these results, T002078 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals of the United Nations (2015) and Regulation (EC) No 1272/2008 on classification, labelling and packing of substances and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute Oral Toxicity:

An acute oral toxicity study with T002078 according to the acute toxic class method in female Crl:WI (Han) (SPF) rats (OECD guideline 423 and EU Method B.1 tris) was performed (Latour, 2016). The substance was administered to six female Wistar rats at 2000 mg/kg body weight. The substance was formulated in propylene glycol (spec. gravity 1.036) at a concentration 200 mg/mL. The rats received a single oral dose of test item, and were observed during 14 days following administration. Mortality/viability was observed twice daily, and bodyweight was monitored at day 1 (prior to administration), 8 and 15. Clinical signs were observed at periodic intervals on the day of dosing (day 1) and once daily thereafter. Necropsy of survivors and macroscopic examination were also performed. Internal macroscopic abnormalities were recorded.

No mortality occurred. Clinical signs observed were hunched posture, piloerection and uncoordinated movements and were noted for all animals between Days 1 and 4. The body weight gain shown by most of the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of all animals.

The oral LD50 value of T002078 in Wistar rats was established to exceed 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Based on these results, T002078 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals of the United Nations (2015) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Acute Inhalation Toxicity:

No reliable acute toxicity study via inhalation were available. However, this endpoint is waived as specific data are available for the oral exposure route.

Acute Dermal Toxicity:

An acute dermal toxicity study does not have to be performed based on the following justification:

• The substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route: LD50>2000 mg/kg bw (OECD 423 study) and no clinical signs observed leading to STOT SE classification. In addition, no abnormalities were found at macroscopic post mortem examination of all animals.
• No systemic effects have been observed in in vivo studies with dermal exposure:
• Skin irritation: not irritating to the skin in an OECD 404 test
• Skin sensitization: the substance was not regarded as a skin sensitizer in a test comparable to the OECD 429 Guideline. No signs of systemic toxicity were noted for the animals in the pre-screen test and no mortality occurred and no clinical signs of systemic toxicity were observed in the animals of the main study.

Justification for classification or non-classification

Acute Oral Classification:

Based on the results showing an acute oral LD50 >2000 mg/kg bw and according to the criteria laid down in Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, T002078 should not be classified for acute dermal toxicity.

Acute Inhalation Classification:

No data were available to decide on the classification for the inhalation route.

Acute Dermal Classification:

No data were available to decide on the classification for the dermal route.