Denatonium benzoate

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

data is from experimental reports following standard procedures

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPP 81-2 (Acute Dermal Toxicity)

Principles of method if other than guideline:

Acute dermal toxicity study of denatonium benzoate (3734-33-6) was performed in rats.

GLP compliance:

yes (incl. QA statement)

Test type:

other: limit test

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Denatonium benzoate

EC Number:

223-095-2

EC Name:

Denatonium benzoate

Cas Number:

3734-33-6

Molecular formula:

C21H29N2O.C7H5O2

IUPAC Name:

N-benzyl-2-[(2,6-dimethylphenyl)amino]-N,N-diethyl-2-oxoethanaminium benzoate hydrate

Test material form:

solid: granular

Details on test material:

- Name of test material: Denatonium benzoate
- Molecular formula: C21H29N2O.C7H5O2
- Molecular weight : 446.58 g/mol
- Substance type: organic
- Physical state: Solid
- SMILES: CCN{+}(CC)(Cc1ccccc1)(CC(=O)Nc1c(C)cccc1C).O{-}C(=O)c1ccccc1

Specific details on test material used for the study:

- Name of test material (as cited in study report): N-benzyl-2-[(2,6-dimethylphenyl)amino]-N,N-diethyl-2-oxoethanaminium benzoate hydrate
-Common name: Denatonium benzoate
- Molecular formula: C21H29N2O.C7H5O2
- Molecular weight: 446.58 g/mol
- Substance type: organic
- Physical state: Solid
- Form: white granular solid
- batch no:22362
- Storage conditions: room temperature, room temperature in the dark

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Details on test animal
TEST ANIMALS
- Source: Harlan U.K Ltd., Blackthron, Bicester, Oxon, U.K.
- Age at study initiation:10-14 weeks old
- Weight at study initiation: male 222-251g and female 202-232g
- Fasting period before study: Rats were not fasted prior to dosing
- Housing: The animals were housed in suspended polypropylene cage furnished with wood flakes. The animals were housed individually during the 24-hr exposure period and in group of 5 , by sex for reminder of the study.
- Diet (e.g. ad libitum): Fed ad libitum (Rat and mouse Expanded Diet no. 1, Special dies services Limited, Witham , Essex, U.k)
- Water (e.g. ad libitum):Drinking water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):19-22°C
- Humidity (%):47-53%
- Air changes (per hr):15 changes per hour
- Photoperiod (hrs dark / hrs light): lighting was controlled by a time switch to give 12hr continuous light and 12hr darkness.
IN-LIFE DATES: From: To:

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Details on exposure
VEHICLE:
- Concentration in vehicle:2000mg
- Amount of vehicle (if gavage): no data available
- Justification for choice of vehicle: no data available
- Lot/batch no. (if required): no data available
- Purity: no data available
MAXIMUM DOSE VOLUME APPLIED: no data available
DOSAGE PREPARATION (if unusual): no data available
CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:

Duration of exposure:

24hr

Doses:

2000mg/kg

No. of animals per sex per dose:

5 male and 5 female

Control animals:

not specified

Details on study design:

Details on study design
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for death or overt signs of toxicity ½, 1,2, and 4hr after dosing and subsequently once daily for 14 days .
Individual body weights were recorded prior to application of the test material on day 0 and on days 7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathological examination was performed. The appearance of any macroscopic abnormalities was recorded.

Statistics:

No data available

Results and discussion

Preliminary study:

No data available

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

There were no death during study

Clinical signs:

other: No signs of systemic toxicity were noted during the study. No signs of skin irritation were noted during the study in nine animals. Necrosis, well- defined erythema, very slight oedema and a hardened dark brown/ black colored scab or sunken scab resemblin

Gross pathology:

No abnormalities were noted at necropsy

Other findings:

No data available

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

LD50 was considered to be >2000mg/kg body weight. When rats were treated with denatonium benzoate (3734-33-6) by dermal application.

Executive summary:

In acute dermal toxicity study male and female Sprague – Dawley rats were treated withdenatonium benzoate(3734-33-6).The animals were housed in suspended polypropylene cage furnished with wood flakes. The animals were housed individually during the 24-hr exposure period and in group of 5 , by sex for reminder of the study. Fed ad libitum (Rat and mouse Expanded Diet no. 1, Special dies services Limited, Witham , Essex, U.k)and Drinking water ad libitum. Acclimation period was 5 days , Rats were not fasted prior to dosing. On the day before treatment the back and flanks of each animals were clipped free of hair using veterinary clippers to expose a skin area of approximately 5cmx 7cm.A group of 5 male and 5 female rats were treated with the test material at dose concentration of 2000mg/kg. The appropriate amount of the test material, as received, pre- weighed into a plastic tube, was applied uniformly to an area of shorn skin approximating to 10% of the total body surface area which had previously been moistened with distilled water .A piece of surgical gauze measuring 7 cm x4 cm was placed over the treatment area and semi occluded with a piece of self-adhesive bandage. The bandage was further secured with a piece of BLENDERM wrapped around each end. The animals were caged individually for the 24hr exposure period. Shortly after dosing the dressing were examined to ensure that they were securely in place. The animals were observed for death or overt signs of toxicity ½, 1,2, and 4hr after dosing and subsequently once daily for 14 days .Individual body weights were recorded prior to application of the test material on day 0 and on days 7 and 14 gross pathological examination was performed. The appearance of any macroscopic abnormalities was recorded.

There was no death during study.No signs of systemic toxicity were noted during the study. No signs of skin irritation were noted during the study in nine animals. Necrosis, well- defined erythema, very slight oedema and a hardened dark brown/ black colored scab or sunken scab resembling a crater were noted at the remaining test site during the study. Scab lifting to reveal dried blood and scab lifting to reveal further bleeding were also noted and an accurate evolution of the degree of erythema and/or oedema was not possible from day 5 onwards at this treatment site.All animals showed expected gain in bodyweight. No abnormalities were noted at necropsy. HenceLD50 was considered to be >2000mg/kg body weight. When rats were treated with denatonium benzoate(3734-33-6)by dermal application.