Reaction mass of 2- (palmitoylamino)ethyl acrylate and 2-(stearoylamino)ethyl acrylate

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 March 2021-April 26 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2- (palmitoylamino)ethyl acrylate and 2-(stearoylamino)ethyl acrylate

EC Number:

950-492-4

Cas Number:

2361378-62-1

Molecular formula:

C21-23H39-43NO3

IUPAC Name:

Reaction mass of 2- (palmitoylamino)ethyl acrylate and 2-(stearoylamino)ethyl acrylate

impurity 1

Reference substance name:

Amides, C16-18, N-(hydroxyethyl)

EC Number:

309-819-0

EC Name:

Amides, C16-18, N-(hydroxyethyl)

Cas Number:

101226-97-5

IUPAC Name:

Amides, C16-18, N-(hydroxyethyl)

impurity 2

Reference substance name:

Zirconium compounds

IUPAC Name:

Zirconium compounds

Test material form:

solid: granular

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Provided by Sponsor, Lot number: 91112Y
- Purity.: 92.4%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
The test substance was put into an air-tight container and stored in test substance storage room at room temperature(acceptable range：from 10℃-30℃) .
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: stability


- Treatment of test material prior to testing (e.g. warming, grinding): The test item was crushed and corrected for purity.

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan(Hino Breeding Center)
- Age at study initiation: nine weeks old at the application of the range-finding study and the main study
- Weight at study initiation: 223.0 g in the range-finding study and 244.8 and 233.4 g in the main study
- Housing: The animals were housed individually after the group allocation and housed with two or three animals per cage after test substance was removed in the main study. The animals were housed in the barrier-system animal rooms in stainless steel cages with mesh floor.
- Diet: Free access to pelleted die t(MF, lot# 201008 and 201214, Oriental Yeast)
- Water: Chlorinated water in which chloric level maintained from 3 to 5 ppm by adding sodium hypochlorite (Purelox) to Hita City supply water was used as drinking water (ad libitum)
- Acclimation period: six days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 25℃
- Humidity (%): 40 to 70%
- Air changes (per hr): 10 to15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light per day

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Remarks:

Purified water

Details on dermal exposure:

TEST SITE
- Area of exposure: back of the trunk of the animal
- % coverage: 5*10 cm area
- Type of wrap if used: The nonwoven fabric was covered and fixed with an elastic adhesive bandage (Silkytex, lot number 10114202, ALCARE)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The elastic adhesive bandage and the nonwoven fabric were removed from the animal and the application site was wiped with an absorbent cotton containing purified water to remove the residual test substance.
- Time after start of exposure: After 24 hours of application，

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Range finding study: animal number 1 = 0.48312; Mian study: animal number 2 = 0.53044g and animal number3 = 0.50546g (2000 mg/kg bw)
- For solids, paste formed: The test substance was put on a nonwoven fabric (5x5cm, lot number 201509151, Ci Medical), moistened with 1mL of purified water and applied to the clipped area.

VEHICLE
- Amount(s) applied (volume or weight with unit): 1 mL
- Lot/batch no. (if required): PC201111 (Takasugi Pharma)

Duration of exposure:

24h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Range-finding: 2000mg/kg bw; 1 animal
Main study: 2000mg/kg bw; 2 animals

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
In the range-finding study and main study, the animals were observed continuously for 10 minutes after the application and observed 30 minutes and three hours after the application. From the day after the application (1 day after the application) to 14 days after the Application, the animals were observed once in the mooring. One day after the application， the animals were observed after the elastic adhesive bandages and the nonwoven fabrics were removed. Body weights were measured on the application day (before application) and 7 and 14 days after the application with an electric balance (SARTORIUS) and body weight changes were calculated.

- Necropsy of survivors performed: The animals were subjected to a gross necropsy 14 days after the application after the general clinical observation. The animals were euthanized by bleeding from the abdominal aorta under isoflurane anesthesia. Application site, external surface of the body, all orifices, subcutis, cranial, thoracic, abdominal and pelvic cavities with their contents were observed.

Results and discussion

Preliminary study:

No mortality or moribundity, no body-weight abnormalities, no macroscopic findings abnormalities occurred in the range-finding study at 2000 mg/kg.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortalities in the main study

Clinical signs:

other: Decreased spontaneous locomotion was observed in all animals from immediately after the application to three hours after the application but disappeared one day after the application and no abnormalities were observed in any animals thereafter.

Gross pathology:

No abnormalities in the main study

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In an acute dermal toxicity study in female Crj:CD(SD) rats, the LD50 was >2000 mg/kg bw.

Executive summary:

In an acute dermal toxicity test (A18-0086), 2 groups of female Crj:CD(SD) rats (1/group in range finding; 2/group in main study) were dermally exposed (occlusive) to the test item in purified water at 2000 mg/kg bw. Animals were observed for 14 days.

Dermal LD50 (Females) = >2000 mg/kg bw

The test item applied at a dose of 2000 mg/kg bw did not cause death of any animals. There were no effects on body weight throughout the study. Decreased spontaneous locomotion was observed in all animals from immediately after the application to three hours after the application but disappeared one day after the application and no abnormalities were observed in any animals thereafter. No macroscopic changes were diagnosed during pathological examination.