Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro-1H-imidazole-1-ethanol (1:1)

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2021/06/26 to 2022/01/20

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro-1H-imidazole-1-ethanol (1:1)

EC Number:

271-272-8

EC Name:

Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro-1H-imidazole-1-ethanol (1:1)

Cas Number:

68527-63-9

Molecular formula:

C22H42N2O.C4H10O4S

IUPAC Name:

2-{2-[(8E)-heptadec-8-en-1-yl]-4,5-dihydro-1H-imidazol-1-yl}ethan-1-ol diethyl sulfate

Test material form:

liquid

Details on test material:

Test item formulations were found to be homogeneous and stable up to 24 hour in vehicle corn oil.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- batch number of test material: 10419
- Purity, including information on contaminants, isomers, etc.: 93 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Dry and dark at ambient room temperature (20 - 30 °C)

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): test item was dispensed under sterile conditions in sterilized glassware

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: In house bred at sa-FORD, Maharashtra, India.
CPCSEA Registration 1256/PO/RcBi/S/09/CPCSEA

- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 - 10 weeks
- Weight at study initiation:
male: minimum: 226 g, maximum 232 g
female: minimum 205 g, maximum 210 g

- Fasting period before study: no
- Housing: all animals were housed individually in polycarbonate cages provided with sterilized corn cobs (M/S. Krishna Corncob Industries- Aurangabad) Batch No.: 221

- Diet (e.g. ad libitum): all animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum, Batch No.: 040521

- Water (e.g. ad libitum): aqua guard filtered tap water was provided ad libitum via drinking bottles. Samples of drinking water was tested to microbial tests and chemical contaminant analysis on twice in year.

- Acclimation period: all animals were acclimatized to the test conditions for 7 days prior to the application of the test item

ENVIRONMENTAL CONDITIONS
- Temperature (°C): min.21.2 °C; max. 22.7 °C
- Humidity (%): min. 53.1 %; max. 62.9 %
- Air changes (per hr): more than 12
- Photoperiod (hrs dark / hrs light): 12 hrs dark and 12 hrs light

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: clipped dorsal intact area of rat's trunk skin
- % coverage: 10
- Type of wrap if used: porous gauze dressing (greater than 10% of body surface area of rat) and non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, by distilled water

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
male: 0.45/0.46 ml
female: 0.4/0.41 ml

- Constant volume or concentration used: yes

Duration of exposure:

24 hrs

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Five male and five female

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
After Test Item application, individual animals were frequently observed approximately at 30 minutes 1, 2, 3, 4, 5 and 6 hours post test item application at day 0 (day of dosing) subsequently all animals were observed twice a day during the 14-day observation period, after dosing. Animals were observed twice daily for any mortality and morbidity during the experimental period. After removal of the patch, all animals were examined for signs of erythema and oedema as per Draize Criteria (Refer Appendix 1) and scored at 24, 48 and 72 hours post dosing.
All rats were weighed on test days 0 (prior to application), 7 and 14.



- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes
- Other examinations performed: clinical signs, body weight, gross pathology

Statistics:

No statistical analysis was performed for LD50 calculation since the study was terminated with limit test.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality and morbidity were observed at limit dose of 2000 mg/kg body weight of test item during the 14 days observation period.

Any other information on results incl. tables

Clinical observation

No systemic reactions of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period.

 

Skin Reaction

Well defined erythema and slight oedema (edges of area well defined by definite raising) were observed in all the animals (animal no. 1 to 10). At 48 & 72 hours observation in animal number 01, 02, 03, 06, 09 and 10 well defined erythema and moderate oedema was observed and animal number 04, 08 showed well defined erythema and slight oedema whereas animal number 05 and 07 showed moderate to severe erythema & slight oedema.

The individual mean score (at 24, 48 and 72 hours)

• of erythema formation for animal no. 01, 02, 03, 04, 05, 06, 07, 08, 09 and 10 at treated sites were 2.00, 2.00, 2.00, 2.00, 2.67, 2.00, 2.67, 2.00, 2.00, 2.00


• and for oedema 2.67, 2.67, 2.67, 2.00, 2.00, 2.67, 2.00, 2.00, 2.67 and 2.67 respectively

 

Mortality and morbidity

No mortality and morbidity were observed at limit dose of 2000 mg/kg body weight of test item during the 14 days observation period.

 

Body weight

The body weight of the animals was within the range of commonly recorded for this strain and age. All rats showed a normal increase in their body weights during the 14 days observation period when compared to day 0.

 

Pathology

External Observation

The external observation of all terminally sacrificed animals did not show any pathological abnormality.

 

Internal observation

The internal observation of all terminally sacrificed animals did not show any pathological abnormality.

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the conditions of this study the acute dermal toxicity LD50 value Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro-1H-imidazole-1-ethanol (1:1) in Wistar rats was estimated as being greater than 2000 mg/kg body weight in male and female Wistar rats.
Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro-1H-imidazole-1-ethanol (1:1) is classified as follows:

- "Category 5/ Unclassifed" (the dermal LD50 range of 2000 to 5000 mg/kg bw) as per Globally Harmonized System of Classification and Labelling of Chemicals (GHS); as there were no mortalities observed at 2000 mg/kg body weight.
- "Category 4" (the dermal LD50 range of 1000 < ATE 2000 mg/kg bw) as per Guidance to Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) of substances and mixtures.

Executive summary:

The acute dermal toxicity study of Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro1 H-imidazole-1-ethanol (1:1) in Wistar Rats, was performed as per the OECD Guideline No. OECD No. 402 and EU Method B.3. 

A starting dose of 2000 mg/kg body weight (bw) based on the test item density (1.0124 g/mL) was applied based on latest body weight of individual animal weight by single dermal application and observed for 14 days after dermal treatment. 

No mortality was observed in any animal till the end of the experimental period.

At 2000 mg/kg body weight, all animals were normal throughout the experimental period. Therefore, the test was terminated with a limit test at 2000 mg/kg body weight. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Under the conditions of this study the acute dermal toxicity LD50 value of Diethyl sulphate, compound with 2-(heptadec-8-enyl)-4,5-dihydro1 H-imidazole-1-ethanol (1:1) in Wistar rats was estimated as being greater than 2000 mg/kg body weight in male and female Wistar rats.