Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 December 2017 - 01 January 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
no

Test material

1
Chemical structure
Reference substance name:
N,N'-(methylenedi-p-phenylene)bis(aziridine-1-carboxamide)
EC Number:
231-034-6
EC Name:
N,N'-(methylenedi-p-phenylene)bis(aziridine-1-carboxamide)
Cas Number:
7417-99-4
Molecular formula:
C19H20N4O2
IUPAC Name:
N,N'-(methylenedi-p-phenylene)bis(aziridine-1-carboxamide)
Test material form:
solid

Test animals

Species:
rat
Strain:
Wistar
Remarks:
RccHan: WIST
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation, India
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: 219.8 to 226.0 g
- Fasting period before study: No
- Housing: 3 females in polypropylene rat cages covered with stainless steel grid top. Autoclaved clean rice husk was used as the bedding material. Wooden blocks were provided as enrichment material.
- Diet (e.g. ad libitum): Ad libitum. Teklad Certified Global High Fiber Rat/Mice Feed manufactured by Envigo, USA. The quality of feed is regularly monitored at Jai Research Foundation.
- Water (e.g. ad libitum): Ad libitum. UV sterilized water filtered through Reverse Osmosis water filtration system. The quality of water is regularly monitored at Jai Research Foundation.
- Acclimation period: 6 to 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23°C
- Humidity (%): 49 to 66%
- Air changes (per hr): At least 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light (06.00 to 18.00 h) and 12 hours darkness.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure:
- % coverage: at least 10%
- Type of wrap if used: porous gauze dressing (not more than 8 ply) and a non-irritating tape (Medi tape 330 hypo-allergenic surgical tape)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cotton soaked in RO water (freshly collected)
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 200, 1000 and 2000 mg/kg body weight
- Concentration (if solution): N/A
- Constant volume or concentration used: N/A
- For solids, paste formed: no
Duration of exposure:
24 h
Doses:
Range finding study: 200, 1000 and 2000 mg/kg body weight
Main study: 2000 mg/kg body weight
No. of animals per sex per dose:
Range finding study: 1 female per dose
Main study: 2 females per dose
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: toxicity and mortality at 0.5, 2, 4 and 6 h post-dermal application on the day of dosing (day 0). Morbidity and mortality twice a day. Erythema and oedema were recorded at 24 h, 48 h and 72 h post patch removal. The clinical signs were recorded once a day. Individual body weight was recorded prior to dermal application on day 0 and on days 7 and 14 post dermal application.
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical observations: evaluation of skin and fur, eyes and mucous membranes, respiratory and circulatory effects, autonomic effects and central nervous system effects, behavioural pattern, somatomotor activity and observation of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study.
Clinical signs:
No clinical signs related to the administration of the test item were observed.
Erythema and oedema was not evident at 24, 48 and 72 h post patch removal in all the rats.
Body weight:
Body weight of all the rats increased on days 7 and 14 when compared to day 0 body weight except of rat N˚ 5 which decreased on days 7 and 14 when compared to day 0 body weight. However, the body weight of rat N˚ 5 increased on day 14 when compared to day 7 (see table 1 below).
Gross pathology:
The macroscopic examinations of the animals at the end of the study did not reveal treatment-related changes.

Any other information on results incl. tables

Table 1: Individual and Mean Body Weight (g) and Body Weight Change (%)

 

Dose

(mg/kg body weight)

Date and

Time of Dosing

Animal N°

Volume

 of

test itemApplied (mL)

Body Weights (g) on Day

Percent Body Weight Change on Day

0

7

14

7

14

200

December 07, 2017 and

10:15 am

1

43.96

219.8

235.0

250.3

6.9

13.9

1000

December 11, 2017 and

10:20 am

2

226.0

226.0

232.9

236.3

3.1

4.6

2000

December 13, 2017 and

10:35 am

3

451.8

225.9

226.9

232.0

0.4

2.7

December 15, 2017 and

10:40 am

4

447.4

223.7

231.7

240.1

3.6

7.3

December 18, 2017 and

10:45 am

5

441.0

220.5

210.0

212.1

-4.8

-3.8

Applicant's summary and conclusion

Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Remarks:
EU criteria.
Conclusions:
The LD50 of the test item is higher than 2000 mg/kg body weight by dermal route in rats.

Executive summary:

The acute dermal toxicity of the test item was studied according to OECD 402 (GLP study). 5 female Wistar rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted sequentially at doses of 200, 1000 and 2000 mg/kg bw in one animal per dose. Based on the results of range finding two additional rats were tested sequentially at a dose of 2000 mg/kg bw. At the end of the 24 h exposure period, the residual test item was removed with cotton soaked in RO water.The skin reactions of each rats were observed at 24, 48 and 72 h post patch removal.The rats were observed for a period of 14 days. There were no treatment-related mortality, clinical signs or necropsy findings recorded. Body weight gain was normal for all rats except for one rat whose body weight decreased on days 7 and 14 compared to day 0 but increased from day 7 to 14. Erythema and oedema was not evident at 24, 48 and 72 h post patch removal in all the rats. Based on the results of the study, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.