oligomeric reaction product of 2-hydroxybenzaldehyde with 1-chloro-2,3-epoxypropane and phenol

Endpoint summary

Description of key information

Under the conditions of this study, the acute oral LD50 value of the test item EPICLON EXA-7250 was found to be above 2000 mg/kg bw in female CRL:(WI) rats.

The median lethal dose (LD50) of EPICLON EXA-7250 after a single dermal administration was found to be greater than 2000 mg/kg bw in male and female Crl:WI rats.

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:

01 September 2015-16 September 2015

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:

yes

Remarks:

These deviations have no presumed effect on the outcome or validity of the study.

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:B006
- Expiration date of the lot/batch:02 March 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature 15-25°C, below 70 RH%

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item of a suitable chemical purity together with all precautions required in the
handling and disposal of the test item were supplied by the Sponsor. The identification
of the test item was made in the Pharmacy of CiToxLAB Hungary Ltd. on the basis of
the information provided by Sponsor.

The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle,
in the Pharmacy of CiToxLAB Hungary Ltd. on the day of administration. The
formulation container was magnetic stirred continuously up to the end of dose
administration procedures.

Species:

rat

Strain:

other: CRL:(WI) rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D- 97633 Sulzfeld.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young healthy adult rats, 8 weeks old
- Weight at study initiation: 172 – 190 g
- Housing: 3 animals / cage
-Fasting: 1 day
- Diet (e.g. ad libitum): Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany (Batch no.: 930 3907, expiry date: December 2015), ad libitum, except for the night before treatment. Tap water from the municipal supply, as for human consumption from a 500 ml bottle, ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Acclimation period: 5-6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.7 – 26.0°C
- Humidity (%): 28 – 70 %
- Air changes (per hr): 15 – 20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From:01 September 2015 To:16 September 2015

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle.
- Amount of vehicle (if gavage):10 mL/kg bw
- Lot/batch no. (if required): SZBE0310V

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose. Initially, 3 female animals were treated with 2000 mg/kg bw of EPICLON EXA-7250. No mortality was observed, therefore further 3 animals were treated at the dose level of 2000 mg/kg bw. As no mortality was observed in this second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris).

Doses:

Initially, 3 female animals were treated with 2000 mg/kg bw of EPICLON EXA-7250. No mortality was observed, therefore further 3 animals were treated at the dose level of 2000 mg/kg bw. As no mortality was observed in this second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris).

No. of animals per sex per dose:

6 animals, 3 animals/group

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter.

- Necropsy of survivors performed: Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthanimal® 40 % inj.; Lot No.: 1409236-06, Expiry Date: September 2017, Produced by: AlfasanNederland BV, Kuipersweg 9, Woerden). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were
observed. Macroscopic abnormalities were recorded.

- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

EPICLON EXA-7250 did not cause mortality at a dose level of 2000 mg/kg bw.

Clinical signs:

other: Acute oral administration of EPICLON EXA-7250 did not cause any test item related effect.

Gross pathology:

There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item EPICLON EXA-7250 was found to be above 2000 mg/kg bw in female CRL:(WI) rats.
According the GHS criteria, EPICLON EXA-7250 can be ranked as "Category 5" or “Unclassified” for acute oral exposure.

Executive summary:

The single-dose oral toxicity of EPICLON EXA-7250 was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in CRL:(WI) rats.

Two groups of 3 female CRL:(WI) rats were treated with the test item at a dose level of 2000 mg/kg bw (Group 1 and Group 2).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was administered formulated in DMSO at a concentration of 200 mg/mL at

a dose volume of 10 mL/kg bw.

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no

further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a

macroscopic examination.

The results of the study were summarized as follows:

Mortality

EPICLON EXA-7250 did not cause mortality at a dose level of 2000 mg/kg bw.

Clinical Observations

Acute oral administration of EPICLON EXA-7250 did not cause any test item related effect.

Body Weight and Body Weight Gain

There were no effects on body weights or body weight gains that could be attributed to treatment with of EPICLON EXA-7250.

Macroscopic Findings

There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

Conclusion:

Under the conditions of this study, the acute oral LD50 value of the test item EPICLON EXA-7250 was found to be above 2000 mg/kg bw in female CRL:(WI) rats. According to the GHS criteria, EPICLON EXA-7250 can be ranked as "Category 5" or “Unclassified” for acute oral exposure.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

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:

02 June 2016- 21 June 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

These deviations are considered to have no impact on the outcome of the study or on the interpretation of the results.

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:B006
- Expiration date of the lot/batch:02 March 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:Controlled Room Temperature (15-25oC, below 70 % RH)


FORM AS APPLIED IN THE TEST: yes

Species:

rat

Strain:

other: Crl:WI rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats Males: 7 weeks; Females: 8-9 weeks
- Weight at study initiation: Males: 215-234 g; Females: 214-237 g
- Fasting period before study: 24 hours
- Housing:Individual caging, Type II. polypropylene/polycarbonate cages. Lignocel 3/4 S, produced by J. Rettenmaier & Söhne GmbH+Co. KG Holzmühle 1, D-73494 Rosenberg, Germany, as certified bedding for laboratory animals was available to rats during the study.
- Diet and water (e.g. ad libitum): Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D- 59494, Soest, Germany (Batch no.: 278 5652, Expiry date: November 2016), ad libitum, and tap water from municipal supply, as for human consumption was provided from 500 ml bottles, ad libitum. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 – 24.9 °C
- Humidity (%): 41 – 69 %
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From:02 June 2016 To: 21 June 2016

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
The back of the animals were shorn (approximately 10% area of the total body surface) approximately 24 hours prior to the treatment. Only the animals without injury or irritation on the skin were used in the test. On the test day (Day 0), the test item was applied as a single dose of 2000 mg/kg
bw after the moistening with sufficient water, applied uniformly over the skin by use of a gauze pad (ca. 5 cm x 5 cm), and remained on the skin throughout an approximately 24-hour exposure period. Sterile gauze pads were placed on the skin of rats at the site of application. These gauze pads were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for approximately 24 hours. At the end of the exposure period, residual test item was removed, using body temperature water.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, residual test item was removed, using body temperature water.
- Time after start of exposure: 24 hours.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):Single dose of 2000 mg/kg bw

Duration of exposure:

24 Hours

Doses:

Single dose of 2000 mg/kg bw

No. of animals per sex per dose:

5 Females per dose
5 Males per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: A clinical examination was performed on the day of treatment, at 2 and 5 hours after the application of the test item, and once each day for 14 days thereafter.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behaviour pattern. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

The body weights were recorded on Day 0 (before test item administration) and on Days 7 and 14 just before necropsy.

Macroscopic examination was performed on all animals. All animals were anaesthetised with pentobarbital sodium (details in 3. 3) and exsanguinated. After
examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred after approximately 24-hour dermal exposure to EPICLON EXA-7250 in Crl:WI rats.

Clinical signs:

other: Each rat was symptom-free during the entire study. No local dermal signs were recorded after treatment with the test item during the 14 days observation period.

Gross pathology:

No external or internal macroscopic findings were observed at a dose level of 2000 mg/kg bw at necropsy.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The median lethal dose (LD50) of EPICLON EXA-7250 after a single dermal administration was found to be greater than 2000 mg/kg bw in male and
female Crl:WI rats. According to the GHS criteria, EPICLON EXA-7250 can be ranked as "Category 5" or "Unclassified" for acute dermal exposure.

Executive summary:

The purpose of the study was to assess the acute dermal toxicity of EPICLON EXA-7250 when administered to rats by a single semi-occlusive dermal application, followed by an observation period of 14 days. The test item was applied dermally to ten (5 males and 5 females) Crl:WI rats as supplied by the Sponsor. A limit test was carried out at 2000 mg/kg body weight (bw) in both

sexes.

Clinical observations were performed on all animals at 2 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Rats were euthanized and subjected to a gross

macroscopic examination at the end of the 2-week observation period (Day 14).

The results of the study were summarized as follows:

Mortality

No mortality occurred after approximately 24-hour dermal exposure to EPICLON EXA-7250 in Crl:WI rats.

Systemic clinical signs

Each rat was symptom-free during the entire study.

Local dermal signs

No local dermal signs were recorded after treatment with the test item during the 14 days observation period.

Body weight and body weight gain

There were no treatment related changes in the body weights. The body weights of the animals were within the range commonly recorded for this strain and age.

Necropsy

No external or internal macroscopic findings were noted at a dose level of 2000 mg/kg bw at necropsy.

Conclusions

The median lethal dose (LD50) of EPICLON EXA-7250 after a single dermal administration was found to be greater than 2000 mg/kg bw in male and female Crl:WI rats.

According to the GHS criteria, EPICLON EXA-7250 can be ranked as "Category 5" or "Unclassified" for acute dermal exposure.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

According to the GHS criteria, EPICLON EXA-7250 can be ranked as "Category 5" or “Unclassified” for acute oral exposure.

According to the GHS criteria, EPICLON EXA-7250 can be ranked as "Category 5" or “Unclassified” for acute dermal exposure.