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Toxicological information

Acute Toxicity: dermal

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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-12-27 to 2018-03-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
9 Oct 2017
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
powder
Details on test material:
white or almost white powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
SPF Caw
Sex:
female
Details on test animals and environmental conditions:
Animals
Five Sprague Dawley rats (SPF Caw) supplied by Elevage JANVIER LABS (53940 Le Genest St Isle– France), were used after an acclimatisation period of at least five days. The females were nulliparous and non-pregnant. At the beginning of the study, the female animals of the treated groups were 8 weeks old. They were identified prior to inclusion in the test by means of a numbered ring on the edge of one ear.

Housing
During the treatment, the animals were kept in individual cages. On D1, the animals were put together into their cage. The rats were kept in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains dust free weed shavings which were changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry. The temperature and relative humidity of the main test were controlled to remain within target ranges of 19°C to 25°C and 30% to 70%, respectively. The rate of air exchange was at least ten changes cycles per hour and the lighting was controlled by a time switch to give twelve hours continuous light (07.00 to 19.00) and twelve hours darkness.

Food and drink
Drinking water (tap-water from public distribution system) and foodstuff (ENVIGO 2016) were supplied ad libitum. Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE).

Administration / exposure

Type of coverage:
open
Vehicle:
water
Remarks:
Distilled water
Doses:
Range-finding study:
- 1 animal treated at 200 mg/kg, 1 female rat, Rf2371
- 1 animal treated at 1000 mg/kg, 1 female rat, Rf2384
- 1 animal treated at 2000 mg/kg, 1 female rat, Rf2385

Main study:
Group 2 treated at 2000 mg/kg, 2 females, Rf2404 & Rf2405
No. of animals per sex per dose:
Range-finding study: 1 animal/dose
Main study: 2 animals/dose
Control animals:
no
Details on study design:

Range-finding study
Approximately 24 hours before the treatment, fur was removed from the dorsal area of the trunk of the test animal by clipping. At least 10 per cent of the body surface area was clear for the application of the test item Nicotine bitartrate dihydrate.
The animals of the treated groups received an effective dose of 200 mg/kg, 1000 mg/kg or 2000 mg/kg body weight of the test item Nicotine bitartrate dihydrate. Distilled water was chosen as it produced the most suitable formulation at the requested concentration.
In view to obtain a solution at 200 mg/kg, 0.2002 g of the test item were weighed and distilled water was added to a 10 mL volumetric flask. The preparation was magnetically stirred and by vortex to obtain a colorless solution before administration. In view to obtain a solution at 1000 mg/kg, 1.0009 g of the test item were weighed and distilled water was added to a 10 mL volumetric flask. The preparation was stirred by vortex to obtain a colorless solution before administration.
In view to obtain a solution at 2000 mg/kg, 1.0017 g of the test item were weighed and distilled water
was added to a 5 mL volumetric flask. The preparation was stirred by vortex to obtain a colorless solution before administration. The animal received by topical application, under non occlusive porous gauze dressing (50 mm x 50 mm non-woven swab of 4-layer patch from MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic microporeTM adhesive tape from 3M), an effective dose of 200 mg/kg, 1000 mg/kg or 2000 mg/kg body weight of the test item administered under a volume of 10 mL/kg body weight, during 24 hours. After 24-hour exposure period, the gauze dressings were removed and the treated area was rinse with distilled water.

Main study
Considering the results obtained in the preliminary study, two additional animals were treated at the dose of 2000 mg/kg body weight.
Approximately 24 hours before the treatment, fur was removed from the dorsal area of the trunk of the test animals by clipping. At least 10 per cent of the body surface area was clear for the application of the test item Nicotine bitartrate dihydrate. Distilled water was chosen as it produced the most suitable formulation at the requested concentration.
In view to obtain a solution at 2000 mg/kg, 1.0013 g of the test item were weighed and distilled water was added to a 5 mL volumetric flask. The preparation was stirred by vortex to obtain a colorless solution before administration. Animals from treated group received by topical application, under non occlusive porous gauze dressing (50 mm x 50 mm non-woven swab of 4-layer patch from MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic microporeTM adhesive tape from 3M), an effective dose of 2000 mg/kg body weight administered under a volume of 10 mL/kg body weight, during 24 hours. After 24-hour exposure period, the gauze dressings were removed and the treated area was rinse with distilled water.
Statistics:
n.a.

Results and discussion

Preliminary study:
No mortality was noted in animal treated at the dose of 200 mg/kg body weight.
No systemic clinical sign related to the administration of the test item was observed.
The body weight evolution of the animal remained normal throughout the study.
The macroscopic examination of the animal at the end of the study did not reveal treatment-related
changes.
No mortality was noted in animal treated at the dose of 1000 mg/kg body weight.
No systemic clinical sign related to the administration of the test item was observed.
The body weight evolution of the animal remained normal throughout the study.
The macroscopic examination of the animal at the end of the study did not reveal treatment-related
changes.
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No mortality was noted in animals treated at the dose of 200, 1000 and 2000 mg/kg bw.
Clinical signs:
No systemic clinical sign related to the administration of the test item was observed.
Body weight:
The body weight evolution of the animals remained normal throughout the study at all dose levels.
Gross pathology:
The macroscopic examination of the animal at the end of the study did not reveal treatment-related changes.
Other findings:
2000 mg/kg bw group:
Red secretions around eyes, nose and muzzle were noted five hours post-dose in one animal (1/3) and
were totally reversible on day 2.
Erythema was noted in all treated animals (3/3) at 24 hours post dose and was totally reversible
between days 2 and 3.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The LD50 of the test item Nicotine bitartrate dihydrate is higher than 2000 mg/ kg body weight by dermal route in the rat.
The test item Nicotine bitartrate dihydrate does not have to be classified in accordance with the Regulation EC No. 1272/2008 on classification, labelling and packaging of substances and mixtures.
No signal word or hazard statement is required.
Executive summary:

The test item Nicotine bitartrate dihydrate was applied, as supplied, onto the intact skin of 1 female Sprague Dawley rat at the dose of 200 mg/kg body weight, then to 1 female Sprague Dawley rat at the dose of 1000 mg/kg body and then to a group of 3 female Sprague Dawley rat at the dose of 2000 mg/kg body weight. The experimental protocol was established on the basis of the method as defined in the O.E.C.D. Test Guideline No. 402 dated October 9th, 2017.

No mortality was noted in animal treated at the dose of 200 mg/kg body weight. No systemic clinical sign related to the administration of the test item was observed. The body weight evolution of the animal remained normal throughout the study. The macroscopic examination of the animal at the end of the study did not reveal treatment-related changes. No mortality was noted in animal treated at the dose of 1000 mg/kg body weight. No systemic clinical sign related to the administration of the test item was observed. The body weight evolution of the animal remained normal throughout the study. The macroscopic examination of the animal at the end of the study did not reveal treatment-related changes.

No mortality was noted in animal treated at the dose of 2000 mg/kg body weight. Red secretions around eyes, nose and muzzle were noted five hours post-dose in one animal (1/3) and were totally reversible on day 2. Erythema was noted in all treated animals (3/3) at 24 hours post dose and was totally reversible between days 2 and 3.

The body weight evolution of the animals remained normal throughout the study. The macroscopic examination of the animal at the end of the study did not reveal treatment-related changes.

In conclusion, the LD50 of the test item Nicotine bitartrate dihydrate is higher than 2000 mg/ kg body weight by dermal route in the rat.

The test item Nicotine bitartrate dihydrate does not have to be classified in accordance with the Regulation EC No. 1272/2008 on classification, labelling and packaging of substances and mixtures.

No signal word or hazard statement is required.