Reaction mass of bis(oxiran-2-ylmethyl) terephthalate and tris(oxiran-2-ylmethyl) benzene-1,2,4-tricarboxylate

Endpoint summary

Description of key information

One acute oral toxicity study was performed in rats. The substance was administered by oral gavage in males and females Sprague-Dawley rats.
The LD50 cut off was determined to be 1000 mg/kg,   at the highest level tested of 2000 mg/kg all three animals died.  No death occurred in all six animals treated at the dose level of 300 mg/kg.
One acute dermal toxicity study was performed. Male and female rats (Sprague Dawley) were treated dermally by occlusive application during 24 hours at a dosage of 2000 mg/kg body weight.  The acute dermal median lethal dose (LD50) in rats was found to be greather than 2000 mg/kg. 

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:

2013-12-17 to 2014-02-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP 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)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

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 (lot no. 0801)
- 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 / individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 260913)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Route of administration:

oral: gavage

Vehicle:

other: DMSO and PEG400

Details on oral exposure:

The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals.
Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.
The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.

Doses:

Dose Level
The starting dose was selected to be 2000 mg/kg body weight. Compound-related mortality was recorded for 2 animals of step 1. Based on these results and according to the acute toxic class method regime, a second step was performed at a dose of 300 mg/kg body weight. No compound-related mortality was
recorded for any animal of step 2. Based on these results and according to the acute toxic class method regime, a third step was performed at a dose of
300 mg/kg body weight. No compound-related mortality was recorded for any animal of step 2. Based on these results and according to the acute toxic
class method regime no further testing was required.

No. of animals per sex per dose:

2000: 3
300: 6

Control animals:

no

Details on study design:

Observation Period
The surviving 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.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central
nervous systems and somatomotor activity and behavior pattern were examined. Particular attention was directed to observations of tremor,
convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Pathology
At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial; lot no.:231073 31 July 2017) at a dosage of approximately 8 mL/kg bw.
All animals were subjected to gross necropsy. All gross pathological changes were recorded and in case of findings the tissues were preserved for a possible histopathological evaluation. The preserved tissues of which no histopathological evaluation was made will be discarded 3 months after the release of the final report unless otherwise agreed upon with the sponsor.

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of
the results is not regarded as necessary.

Sex:

female

Dose descriptor:

approximate LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut-off (rat): 1000 mg/ kg bw

Mortality:

Two animals treated with the test item at a dose of 2000 mg/kg bw were found dead on the second day of observation. The remaining animal survived until the end of the study.

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, eyes half closed, catalepsies, kyphosis, bradykinesia and wasp waist. The most relevant clinical sign

Gross pathology:

Macroscopic findings of surviving animals:
At necropsy, no macroscopic findings were observed in any animal of any step.
Macroscopic findings of animals not having survived until the end of the observation period:
Necropsy revealed gaseous distension of stomach, dark blood-red colored duodenum and jejunum.

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Under the conditions of the present study, a single oral application of the test material to rats at a dose of
2000mg/kg body weight was associated with signs of toxicity and mortality.
Under the conditions of the present study, a single oral application of the test material to rats at a dose of 300mg/kg body
weight was associated with signs of toxicity but no mortality.
The median lethal dose of the test material after a single oral administration to female rats, observed over a period of 14 days is:
LD50 (rat) is > than 300 mg/kg and < 2000 mg/kg.
LD50 cut-off (rat): 1000 mg/ kg bw
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test material has obligatory
labelling requirement for toxicity.
According to Annex I of Regulation (EC) 1272/2008 the test material has obligatory labelling requirement for toxicity and is classified into Category 4.
According to GHS (Globally Harmonized Classification System) the test material has obligatory labelling requirement for toxicity and is classified into Category 4.

Executive summary:

Summary Results

One group 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 in the vehicle DMSO and PEG400 (ratio 1:5)at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

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 300 mg/kg body weight. The test item was suspended in the vehicle DMSO and PEG400 (ratio 1:5) at a concentration of 0.03 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. Two animals treated with the test item at a dose of 2000 mg/kg bw were found dead on the second day of observation. The remaining animal survived until the end of the study.

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, eyes half closed, catalepsies, kyphosis, bradykinesia and wasp waist. The most relevant clinical signs persisted on the day of treatment in all animals. The surviving animal recovered from all clinical signs on the fifth day of the observation period.

The most relevant clinical findings in the animals treated with the test item at a dose of 300 mg/kg bw were reduced spontaneous activity, piloerection, eyes half closed, catalepsies, kyphosis, bradykinesia and wasp waist. The most relevant clinical signs persisted on the day of treatment in all animals. All animals recovered from all clinical signs on the third day of the observation period.

Throughout the 14-day observation period, the weight gain of the surviving 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.

Macroscopic findings of animals not having survived until the end of the observation period:

Necropsy revealedgaseous distension of stomach,darkblood-red colored duodenum and jejunum.

 LD₅₀cut-off:                                                       1000 mg/kg bw

Species/strain:                                                  WISTAR Crl: WI(Han) rats

Number of animals:                                         3 per step / 3 steps performed

Vehicle:                                                               DMSO and PEG400 (ratio 1:5)

Method:                                                               OECD 423; EC 440/2008, Method B.1 tris; OPPTS 870.1100

Conclusion

Under the conditions of the present study, a single oral application of the test material to rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity and mortality.

Under the conditions of the present study, a single oral application of the test material to rats at a dose of 300 mg/kg body weight was associated with signs of toxicity but no mortality.

The median lethal dose of the test material after a single oral administration to female rats, observed over a period of 14 days is:

LD₅₀(rat) is > than 300 mg/kg and <2000 mg/kg.

LD₅₀cut-off (rat): 1000 mg/ kg bw

In conformity with the criteria given inAnnex VI to Commission Directive 2001/59/EC the test material has obligatory labelling requirement for toxicity.

According to Annex I of Regulation (EC) 1272/2008 the test material has obligatory labelling requirement for toxicity and is classified into Category 4.

According to GHS (Globally Harmonized Classification System) the test material has obligatory labelling requirement for toxicity and is classified into Category 4.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-01-28 to 2014-03-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental 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 (lot no. 801)
- 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 individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 131113)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

Preparation of the Animals:
The animals were marked for individual identification by tail painting.
Approximately 24 hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper.
Care was taken to avoid abrading the skin, and only animals with healthy intact skin were used.
No less than 10% of the body surface was cleared for the application.
Prior to the application a detailed clinical observation was made of all animals.
Application:
The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface.
The test item was held in contact with the skin by a dressing throughout a 24-hour period. The dressing consisted of a gauze-dressing and
non-irritating tape and was fixed with an additional dressing in a suitable manner.

Duration of exposure:

The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was removed using the vehicle aqua ad injectionem.

Doses:

The test item was applied at a single dose of 2000 mg/kg body weight to each animal.

No. of animals per sex per dose:

5 male and 5 female

Control animals:

not required

Details on study design:

Observation period:
All animals were observed for 14 days after dosing.

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

Clinical Examination:
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.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Pathology:
At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial) at the dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded and in case of findings the tissues were preserved for a possible histopathological evaluation. The preserved tissues of which no histopathological evaluation was made will be discarded 3 months after the release of the final report unless otherwise agreed upon with the sponsor.

Evaluation of Results:
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by sex and dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Sex:

male/female

Dose descriptor:

approximate LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality was observed.

Clinical signs:

other: No treatment-related effects were observed.

Gross pathology:

No treatment-related effects were observed.

Other findings:

No erythema or oedema was observed.

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

Under the conditions of the present study, single dermal application of the test material to rats at a dose of 2000 mg/kg body weight was associated with no mortality and neither signs of toxicity nor signs of irritation.
The dermal LD50 was determined to be > 2000 mg/ kg body weight.
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test material has no obligatory labelling requirement for percutaneous toxicity.
According to Annex I of Regulation (EC) 1272/2008 the test material has no obligatory labelling requirement for percutaneous toxicity and is unclassified.
According to GHS (Globally Harmonized Classification System) the test material has no obligatory labelling requirement for percutaneous toxicity and is not classified.

Executive summary:

Summary Results

LD50: > 2000 mg /kg bw

Species/strain: WISTAR Crl: WI(Han) rats

Vehicle (moistening): aqua ad injectionem

Number of animals: 5 male and 5 female

Duration of exposure: 24 hours

Method: OECD 402, EC 440/2008, Method B.3, OPPTS 870.1200

 

Signs of systemic toxicity related to dose level used, time of onset and duration:

No treatment-related effects were observed.

Effect on organs (related to dose level):

No treatment-related effects were observed.

Signs of irritation:

No erythema or oedema was observed.

Conclusion

Under the conditions of the present study, single dermal application of the test material to rats at a dose of 2000 mg/kg body weight was associated with no mortality and neither signs of toxicity nor signs of irritation.

The dermal LD50 was determined to be > 2000 mg/ kg body weight.

In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test material has no obligatory labelling requirement for percutaneous toxicity.

According to Annex I of Regulation (EC) 1272/2008 the test material has no obligatory labelling requirement for percutaneous toxicity and is unclassified.

According to GHS (Globally Harmonized Classification System) the test material has no obligatory labelling requirement for percutaneous toxicity and is not classified.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

- oral toxicity:

Based on the above stated assessment of the acute oral toxicity of reaction mass of bis(2,3-epoxypropyl) terephthalate and tris(oxiranylmethyl) benzene-1,2,4-tricarboxylate the substance does need to be classified as Acute Oral toxicity Category 4 according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and accordingCLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council)as implementation of UN-GHS in the EU.

- dermal toxicity:

Based on the above stated assessment of the acute dermal toxicity of reaction mass of bis(2,3-epoxypropyl) terephthalate and tris(oxiranylmethyl) benzene-1,2,4-tricarboxylate (absence of toxicity up to 2000 mg/kg) the substance does not need to be classified according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and accordingCLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council)as implementation of UN-GHS in the EU.

- inhalation toxicity:

Due to the very low vapour pressure of the substance, the fact that the substance is imported into the EU in a formulated form as a granulate, the inhalation route of exposure is considered to be unlikely. Therefore no classification for acute inhalation toxicity is deemed necessary accordingto Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and accordingCLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council)as implementation of UN-GHS in the EU.