Distillate of tobacco leaf, Nicotiana tabacum, solanaceae

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2015-05-26 to 2015-08-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was performed in accordance with standard test protocols (OECD 402) in a quality controlled laboratory. The study is valid according to criteria mentioned in the test protocols.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Nicotine

EC Number:

200-193-3

EC Name:

Nicotine

Cas Number:

54-11-5

Molecular formula:

C10H14N2

IUPAC Name:

3-(1-methyl-2-pyrrolidinyl)pyridine

Test material form:

liquid

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

cross-reference: section 7.2.3

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

Preliminary study: 1000, 200, 50 and 5 mg/kg bw dose in females
Main study: 200, 100 and 50 mg/kg bw dose in females and 50 mg/kg bw dose in males

Duration of treatment / exposure:

24-hour exposure period

Observation period:

14-day observation period

Number of animals:

Preliminary study: n=2 animals/dose
Main study: n=5 animals/sex/dose

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Remarks on result:

other: see "irritant/corrosive response data" and for detailed information please see "Any other information on results incl. tables".

Results 2

Irritation parameter:

edema score

Remarks on result:

other: see "irritant/corrosive response data" and for detailed information please see "Any other information on results incl. tables".

Irritant / corrosive response data:

Dermal irritation symptoms were observed on the treatment site in both sexes.
In males treated with 50 mg/kg bw dose (group 1), very slight redness (score 1), well defined redness (score 2) and moderate to severe redness (score 3) was observed in one animal between Day 1 and Day 7. Moderate to severe redness (score 3) and severe redness (score 4) was detected in two animals between Day 1 and Day 7. Very slight redness (score 1) and well defined redness (score 2) was recorded in one animal between Day 1 and Day 7. Very slight redness (score 1) occurred in one animal between Day 1 and Day 5. Another sign as desquamation appeared in four animals on Day 6 and in one animal on Day 7. Crusting was recorded in four animals. It was observable between Day 7 and Day 14. Wound was found in two animals. It was observable between Day 11 and Day 14.
One male became free of skin irritation symptoms by Day 10.
In females treated with 50 mg/kg bw dose (group 1), very slight redness (score 1) was observed in two animals on Day 1. Well defined redness (score 2) and very slight redness (score 1) was detected in one animal on Day 1 and on Day 2, respectively. Moderate to severe redness (score 3) and well defined redness (score 2) was recorded in one animal on Day 1 and on Day 2, respectively.
Very slight redness (score 1) occurred in one animal treated with 100 mg/kg bw dose (group 2) on Day 1.
The survivor females became free of skin irritation symptoms by Day 3.
There was no oedema observed during the 14-day observation period in both sexes, but it was observable in females of group 2 and 3 on necropsy day.

Any other information on results incl. tables

Local symptoms (dermal irritation)

Oe:0 = No oedema

R:0 = No redness

R:1 = Very slight redness

R:2 = Well defined redness

R:3 = Moderate to severe redness

R:4 = Severe redness

C = Crusting; W = Wound; D = Desquamation

Males - Group 1 (50 mg/kg bw)

Animal No.	Day 1	Day 3	Day 7	Day 14
#1	R:1; Oe:0	R:2; Oe:0	R:1; Oe:0; D	R:0; Oe:0; C
#2	R:3; Oe:0	R:4; Oe:0	R:4; Oe:0; D; C	R:0; Oe:0; C
#3	R:1; Oe:0	R:1; Oe:0	R:2; Oe:0; D; C	R:0; Oe:0; W
#4	R:3; Oe:0	R:3; Oe:0	R:4; Oe:0; D; C	R:0; Oe:0; W
#5	R:1; Oe:0	R:1; Oe:0	R:0; Oe:0; D	R:0; Oe:0

Females - Group 1 (50 mg/kg bw)

Animal Number	Day 1	Day 2	Day 3
#1	R:1; Oe:0	R:0; Oe:0	R:0; Oe:0
#2	R:1; Oe:0	R:0; Oe:0	R:0; Oe:0
#3	-	-	-
#4	R:2; Oe:0	R:1; Oe:0	R:0; Oe:0
#5	R:3; Oe:0	R:2; Oe:0	R:0; Oe:0

Females - Group 2 (100 mg/kg bw)

Animal Number	Day 1	Day 2	Day 3
#1	-	-	-
#2	-	-	-
#3	-	-	-
#4	R:1; Oe:0	R:0; Oe:0	R:0; Oe:0
#5	-	-	-

Females - Group 3 (200 mg/kg bw)

No data available due to lethality.

The survivor females were free of skin irritation symptoms from Day 4 until the end of observation period.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant)

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Nicotine caused dermal irritation symptoms on the treatment site in both sexes in an acute toxicity study by dermal route in rabbits exposed to 3.0-3.2 mg/cm². No corrosion symptoms, according to CLP-regulation, Annex I, were observed, thus the substance is classified as Skin Irrit.2

Executive summary:

Data for skin irritation potential were obtained from the acute dermal toxicity test (OECD 402, section 7.2.3.).

The test item caused dermal irritation symptoms on the treatment site in both sexes.

In males, very slight to severe erythema was observed between Day 1 and Day 7 and other signs as desquamation, crusting and wound were detected between Day 6 and the necropsy day, as well.

In females, very slight to severe erythema was recorded in 50 mg/kg bw dose group between treatment day and Day 2. Very slight to severe erythema and slight oedema was found in 100 mg/kg bw dose group between treatment day and Day 1. Moderate to severe erythema and slight to moderate oedema was detected in 200 mg/kg bw dose group on the treatment day.

Justification of the classification category (Skin Irrit. 2) is based on calculation of the amount of test substance per cm² and comparison to the test substance concentration of 80 mg/cm² employed in the EU B.4/OECD TG 404 (according to the recommended approach of testing and assessment strategy for skin corrosion/irritation in the Guidance on Information Requirements and Chemical Safety Assessment Chapter R.7a, Version 4.0, July 2015, Section R.7.2.6.2 (d)):

The back of animals was shaven (approximately 10 % area of the total body surface, approx. 36 cm²) 24 hours prior to the treatment. Group 1 (males) was treated with 50 mg nicotine per kg bw. The body weight range in main study at starting (males, group 1) was 2210 - 2310 g, thus the dose was 110.5-115.5 mg/36 cm². Therefore the animals were exposed to 3.0-3.2 mg/cm² nicotine. Compared to the standard test design treatment of 80 mg/cm², this small amount of nicotine per cm² resulted in clear dermal irritation symptoms on the treatment site in both sexes, especially in males: slight to severe redness, desquamation, crusting and wound. No corrosion symptoms (CLP,Annex I: 3.2.1.1. “irreversible damage of the skin […] typified by ulcers, bleeding, bloody scabs and, by the end of observation at 14 days, by discolouration due to blanching of the skin, complete areas of alopecia, and scars”) were observed, thus the substance is classified as Skin Irrit. 2.