13-Docosenamide, N-[3-(dimethylamino)propyl]-, (13Z)-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From 1st March to 21 April 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was performed similarly to the OECD 406 guideline. No positive control was used in this study and there was no reference to historical positive control data and only 10 and 5 animals were used in the treated and the control groups, respectively. The test was performed on a very similar structure (saturated chain instead of an insaturated chain).

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

no positive control data, only 10 animals in the treated group

Principles of method if other than guideline:

not applicable

GLP compliance:

not specified

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

-

Test material

Test material information

Constituent 1

Reference substance name:

Docosanamide,N-[3-(dimethylamino)propyl]-

IUPAC Name:

Docosanamide,N-[3-(dimethylamino)propyl]-

Constituent 2

Reference substance name:

N-[3-(dimethylamino)propyl]docosanamide

EC Number:

262-134-8

EC Name:

N-[3-(dimethylamino)propyl]docosanamide

Cas Number:

60270-33-9

Molecular formula:

C27H56N2O

Test material form:

other: white powder

Details on test material:

- Name of test material (as cited in study report): 04-S-014
- Storage condition of test material: a dark place at room temperature

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Japan SLC, Inc. Japan
- Age at study initiation: 4 weeks
- Weight at study initiation: 336g
- Housing: group of 5 animals in aluminium cages (W350 x D400 x H230)
- Diet (e.g. ad libitum): solid food (RC4: Oriental YEast Co., Ltd), ad libitum
- Water (e.g. ad libitum): tap water after passing 5µm wartridge filter, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 +/- 2°C
- Humidity (%): 50 +/- 10%
- Air changes (per hr): 17
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: no data

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: intradermal injection: physiological saline (Lot 3I86 Otsuka Pharmaceutical Factory Inc.); contact sensitization and challenge: 50% ethanol in water (Lot. AFE2540 Wako Pure Chemical Industries Ltd.)

Induction 2

Concentration / amount:

intradermal sensitisation: 0.01% test substance in physiological saline or an emulsion of 0.02% test substance in physiological saline and FCA in 1:1
Contact sensitisation: 30%
Challenge: From 0.5 to 30%

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: intradermal injection: physiological saline (Lot 3I86 Otsuka Pharmaceutical Factory Inc.); contact sensitization and challenge: 50% ethanol in water (Lot. AFE2540 Wako Pure Chemical Industries Ltd.)

Challenge 2

Concentration / amount:

intradermal sensitisation: 0.01% test substance in physiological saline or an emulsion of 0.02% test substance in physiological saline and FCA in 1:1
Contact sensitisation: 30%
Challenge: From 0.5 to 30%

No. of animals per dose:

8 animals for the preliminary test
10 for the sensitisation group
5 for the control group

Details on study design:

RANGE FINDING TESTS:
a) 0.1 mL of test substance at concentration comprised between 0.01 and 5% was intradermally injected in the lateral abdomen of 4 animals.
b) closed patches with 0.05 mL of each test sample (6 tested concentrations comprised between 0.3 and 10% in ethanol) were applied on four symetric sites on the back of 4 animals. One week before, these animals were intradermally treated with the test susbtance and FCA). The contact sensitisation was made under occlusive conditions for 24 hours.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 (a, b and c) x2 sites (for intradermal injection)
- Exposure period: d0-d8
- Test groups:
a) an emulsion of FCA and physiological saline in 1:1
b) 0.01% TS in vehicle (physiological saline)
c) 0.02% TS and FCA in 1:1
- Control group: vehicle (physiological saline)
- Site: 2 sites (right and left) on the back
- Frequency of applications: Topical induction 8 days after intradermal one
- Duration: Acute (intradermal) / 48 h (topical)
- Concentration: intradermal injection:0.01% TS, contact sensitisation: 30% TS.

On day 8 from initiation of sensitisation, 0.2 mL of the sample was soaked in lint cloth of 2x4 cm in size and applied as a closed patch for 48 hours using occlusive patch.The control animals were treated similarly using the vehicle (50% ethanol in water). Since the test substance is non-irritating, 0.5 g of 10% SDS was applied on day 7.

B. CHALLENGE EXPOSURE
- No. of exposures: 1x6 sites
- Day(s) of challenge: d21
- Exposure period: 24 hours
- Test groups: TS
- Control group: TS
- Site: lateral abdomen
- Concentrations: 6 different (from 0.5 to 30%): 0.5; 1; 3; 5; 10 and 30%
- Evaluation (hr after challenge): 3, 24 and 48 hours after the patch removal

OTHER: To assess the skin reactions after the closed patch in the preliminary assay and after the challenge in the main study, the Draize scale was followed.

Challenge controls:

not applicable

Positive control substance(s):

no

Remarks:

no positive control was used in the study and no reference to positive historical data was made.

Results and discussion

Positive control results:

not applicable as no positive control was used.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

3

Group:

test chemical

Dose level:

0.5 to 30 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

score for erythema and edema= 0 for all treated animals at any tested concentration

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 3.0. Group: test group. Dose level: 0.5 to 30 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: score for erythema and edema= 0 for all treated animals at any tested concentration.

Results 2

Reading:

2nd reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.5 to 30%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

score for erythema and edema= 0 for all treated animals at any tested concentration

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.5 to 30%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: score for erythema and edema= 0 for all treated animals at any tested concentration.

Results 3

Reading:

other: 3rd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.5 to 30%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

score for erythema and edema= 0 for all treated animals at any tested concentration

Remarks on result:

other: see Remark

Remarks:

Reading: other: 3rd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.5 to 30%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: score for erythema and edema= 0 for all treated animals at any tested concentration.

Results 4

Reading:

1st reading

Hours after challenge:

3

Group:

negative control

Dose level:

0.5 to 30%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

score for erythema and edema= 0 for all control animals at any tested concentration

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 3.0. Group: negative control. Dose level: 0.5 to 30%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: score for erythema and edema= 0 for all control animals at any tested concentration.

Results 5

Reading:

2nd reading

Hours after challenge:

24

Group:

negative control

Dose level:

0.5 to 30%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

score for erythema and edema= 0 for all control animals at any tested concentration

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0.5 to 30%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: score for erythema and edema= 0 for all control animals at any tested concentration.

Results 6

Reading:

other: 3rd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0.5 to 30%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

score for erythema and edema= 0 for all control animals at any tested concentration

Remarks on result:

other: see Remark

Remarks:

Reading: other: 3rd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0.5 to 30%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: score for erythema and edema= 0 for all control animals at any tested concentration.

Any other information on results incl. tables

Results of the preliminary test:

Table 7.4.1/1:result of primary skin irritation after the intradermal administration

At observation		24 hrs after administration							48 hrs after administration							72 hrs after administration
Samples		i	ii	iii	iv	v	vi	vii	i	ii	iii	iv	v	vi	vii	i	ii	iii	iv	v	vi	vii
Animal No.		4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4	4
Judgment	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	±	0	0	0	0	0	0	2	0	0	0	0	0	0	4	0	0	0	0	0	1	4
	+	0	0	0	0	1	2	2	0	0	0	0	1	2	0	0	0	0	0	1	1	0
	++	0	0	0	0	3	2	0	0	0	0	0	3	2	0	0	0	0	0	3	2	0
	+++	4	4	4	4	0	0	0	4	4	4	4	0	0	0	4	4	4	4	0	0	0

i: 5%; ii: 1%; iii: 0.5% ; iv: 0.1% ; v: 0.05% ; vi: 0.03%; vii: 0.01%

-: no reaction; ±: slight erythema; +: apparent erythema; ++: apparent erythema with edema; +++: eschar formation and necrosis

Table 7.4.1/2:result of primary skin irritation after the 24-hour closed patch application

Concentration of TS (%)	Observation time after patch removal (hr.)	Erythema score					Edema score
		0	1	2	3	4	0	1	2	3	4
10	3	0	4	0	0	0	4	0	0	0	0
	24	0	4	0	0	0	4	0	0	0	0
	48	0	4	0	0	0	4	0	0	0	0
5	3	4	0	0	0	0	4	0	0	0	0
	24	4	0	0	0	0	4	0	0	0	0
	48	4	0	0	0	0	4	0	0	0	0
3	3	4	0	0	0	0	4	0	0	0	0
	24	4	0	0	0	0	4	0	0	0	0
	48	4	0	0	0	0	4	0	0	0	0
1	3	4	0	0	0	0	4	0	0	0	0
	24	4	0	0	0	0	4	0	0	0	0
	48	4	0	0	0	0	4	0	0	0	0
0.5	3	4	0	0	0	0	4	0	0	0	0
	24	4	0	0	0	0	4	0	0	0	0
	48	4	0	0	0	0	4	0	0	0	0
0.3	3	4	0	0	0	0	4	0	0	0	0
	24	4	0	0	0	0	4	0	0	0	0
	48	4	0	0	0	0	4	0	0	0	0

 

Main test:

No abnormalities in the general conditions was observed after sensitisation or challenge during the study period and the animals showed satisfactory body weight increases.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, Erucamidopropyl dimethylamine is not sensitising in the guinea-pig maximisation test. Erucamidopropyl dimethylamine is therefore not classified for skin sensitisation according to the Regulation (EC) 1272/2008 (CLP) and the Directive 67/548/EEC.

Executive summary:

In a Guinea-Pig Maximisation Test performed similarly to OECD No. 406 test guideline,Docosanamide,N-[3-(dimethylamino)propyl]-(purity of 99%) was tested for its skin sensitising potential in Albino Hartley female guinea pigs.  

A preliminary test on 4 animals using intradermal injections of the test substance in physiological saline at concentration from 0.01 to 5% showed that higher concentration than 0.01% induced skin irritation with the presence of erythema and edema. The concentration of 0.01% only induced a mild skin irritation. Therefore the intradermal induction concentration was chosen as 0.01% for the main test.

In 4 another animals, several concentrations from 0.3 to 10% were tested by topical application during 24hrs one week after intradermal injection of the test substance in emulsion with Freund’s Complete Adjuvant (FCA). Only the highest concentration (10%) induced a slight skin irritation (erythema score of 1). A range of challenging concentration between 0.5 and 30% was chosen for the main test.

 

For the main test, 5 animals were applied the vehicle (physiological saline) only (control group) and 10 other animals were applied the test substance. For the induction phase, animals received two intradermal injections on day 0 (right and left sites of the back) of the vehicle (physiological saline) in FCA or the test substance at 0.01% in physiological saline or the test substance at 0.01% in FCA emulsion. One week later, as the test substance is non-irritating, 0.5 g of 10% SDS was applied. The next day (Day 8), a second induction was performed by a topical application of the test substance at 30% in ethanol. Two weeks after the topical induction phase (day 21), challenge was performed by applying 6 different concentrations (from 0.5 to 30%) of the test substance in ethanol under occlusive conditions for 24 hours. Observation and grading of skin reactions was performed 3, 24 and 48 hours after patch removal to assess potential sensitisation.

No skin reaction and therefore no sign of sensitisation was observed 3, 24 and 48 hours after the challenge in any animal (control or treated groups).

Therefore, under the conditions of this test,Docosanamide,N-[3-(dimethylamino)propyl]-is not considered as a skin sensitiser according to the criteria of the Regulation (EC) 1272/2008 (CLP) and the Directive 67/548/EEC (DSD). This study is considered as acceptable and satisfies the requirement for skin sensitization endpoint.