Amines, N-(3-aminopropyl)-N'-C12-18-alkyltrimethylenedi-, reaction products with N-C12-18-alkyltrimethylenediamines and formic acid

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2002-08-07 to 2002-09-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

Amines, tallow alkyl[3-(tetrahydropyrimidinyl)propyl]

EC Number:

296-557-4

EC Name:

Amines, tallow alkyl[3-(tetrahydropyrimidinyl)propyl]

Cas Number:

92797-22-3

IUPAC Name:

92797-22-3

Constituent 2

Reference substance name:

Amines, suifalkyl [(tetrahydropyrimidinyl)3-propyl]

IUPAC Name:

Amines, suifalkyl [(tetrahydropyrimidinyl)3-propyl]

Details on test material:

- Name of test material (as cited in study report): POLYRAM L200
- Chemical name : Amines, suifalkyl [(tetrahydropyrimidinyl)3-propyl]
- CAS number : 92797-22-3
- Physical state: clear brown liquid
- Analytical purity: > 95% expressed in amines content
- Purity test date: 2002-06-20
- Lot/batch No.: 94052206
- Expiration date of the lot/batch: July 2003.
- Storage condition of test material: in dark and at room temperature

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Strain and sanitary status: Hartley Crl: (HA) BR, Caesarian obtained, Barrier sustained - Virus Antibody Free (COBS - VAF)
- Source: Charles River Laboratories, L’Arbresle, France.
- Age at study initiation: 1-2 months.
- Weight at study initiation: 373 ± 14 g for the males and 372 ± 15 g for the females.
- Housing: individually in polycarbonate cages.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): ca. 12 cycles/hour of filtered, non-recycled
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: 0.9% NaCl in water

Induction 2

Concentration / amount:

Main study:
-Induction:
intradermal injection: 0.1% w/w in 0.9% NaCl.
topical application: 1% w/w in 0.9% NaCl.
-Challenge:
1% w/w in 0.9% NaCl.

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: 0.9% NaCl in water

Challenge 2

Concentration / amount:

Main study:
-Induction:
intradermal injection: 0.1% w/w in 0.9% NaCl.
topical application: 1% w/w in 0.9% NaCl.
-Challenge:
1% w/w in 0.9% NaCl.

No. of animals per dose:

Preliminary experiment: 6 males and 6 females.
Main experiment: 10 males and 10 females in the treatment group, 5 males and 5 females in the control group.

Details on study design:

RANGE FINDING TESTS:
For both the preliminary test and the main test, the application sites of all animals were:
• clipped before intradermal injections (interscapular region 4 cm x 3 cm),
• clipped before topical applications of the induction phase (same region),
• clipped and shaved before topical applications of the challenge phase
(each flank 3 cm x 3 cm),
• shaved before the 48-hour reading of the challenge phase and before the 24 and
48-hour reading of the induction phase (preliminary test).

Concentrations tested in the range finding test:
By intradermal route (tested concentrations: 10 %, 5 %, 1% and 0.1% (w/w)):
• intradermal injections of the dosage form preparations (0.1 mL) were performed in the
interscapular region,
• cutaneous reactions were evaluated approximately 24, 48 hours and 6 days after the
injections.
By cutaneous route
Under the conditions of the induction phase (tested concentrations: 100%, 50%, 25%, 10%, 5%
and 1 % (w/w)):
• a filter paper (approximately 8 cm2) was fully-loaded with a dosage form preparation and was
then applied to the clipped area of the skin. The filter paper was held in place by means of an
occlusive dressing for 48 hours,
• cutaneous reactions were evaluated 24 and 48 hours after removal of the dressing.
Under the conditions of the challenge phase (tested concentrations: 100%, 50%, 25%, 10%, 5%
and 1 % (w/w)):
• the filter paper of a chamber (Finn Chamber) was fully-loaded with a dosage form
preparation. The chamber was then applied to the clipped area of the skin (one concentration
per flank). The chamber was held in place by means of an occlusive dressing for 24 hours,
• cutaneous reactions were evaluated 24 and 48 hours after removal of the dressings.

Criteria for selection of concentrations
The following criteria were used:
• the concentrations should be well-tolerated systemically and locally,
• intradermal injections should cause moderate irritant effects (no necrosis or ulceration of
the skin),
• cutaneous application for the induction should cause at most weak or moderate skin reactions
or be the maximal practicable concentration,
• cutaneous application for the challenge phase should be the highest concentration which does
not cause irritant effects.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 1 intradermal exposure and 1 epicutaneous (occlusive) exposure
- Exposure period: intradermal on day 1, epicutaneous on day 8
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: the interscapular region, same site for both induction exposures
- Frequency of applications: single treatment for both intradermal epicutaneous exposures
- Duration: intradermal exposure: single injection, epicutaneous exposure: 48h under occlusive dressing
- Concentrations:
intradermally for induction: Freund's complete adjuvant (FCA) diluted at 50% (v/v) with 0.9% NaCl, 0.1% (w/w) test item in 0.9% NaCl and 0.1% in a mixture FCA/0.9% NaCl (50/50, v/v),
epicutaneous for induction: 1% (w/w) in 0.9% NaCl

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 22
- Exposure duration: 24 h under occlusive dressing
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: treatment with test item on the right flank, treatment with vehicle on the left flank
- Concentrations: 1% (w/w) in 0.9% NaCl (0.9% NaCl as control)
- Evaluation (hr after challenge): 24 , 48 hours after removal of the dressing according to the method of Draize (See table 1 in results and discussion free-text for details)

GENERAL
- The animals were observed at least once a day during the study in order to check for clinical signs and mortality.
- The animals were weighed individually on the day of allocation into the groups, on the first day of the study (day 1) and on the last day of the study (day 25).

Challenge controls:

Yes-see above.

Positive control substance(s):

yes

Remarks:

Mercaptobenzothiazole

Results and discussion

Positive control results:

Regular checks were performed at the laboratory with the positive control substance mercaptobenzothiazole . The most recent reliability check achieved a 100% response in the test group and 0% response in the control group at challenge.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

1% (w/w) in 0.9% NaCl

No. with + reactions:

4

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1% (w/w) in 0.9% NaCl. No with. + reactions: 4.0. Total no. in groups: 20.0.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1% (w/w) in 0.9% NaCl

No. with + reactions:

4

Total no. in group:

20

Clinical observations:

dryness of the skin was also observed at the 48-hour reading in the four animals showing cutaneous reactions.

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1% (w/w) in 0.9% NaCl. No with. + reactions: 4.0. Total no. in groups: 20.0. Clinical observations: dryness of the skin was also observed at the 48-hour reading in the four animals showing cutaneous reactions..

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

1% (w/w) in 0.9% NaCl

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 1% (w/w) in 0.9% NaCl. No with. + reactions: 0.0. Total no. in groups: 10.0.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

1% (w/w) in 0.9% NaCl

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1% (w/w) in 0.9% NaCl. No with. + reactions: 0.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

At the 48-hour reading , a discrete erythema (grade 1) was observed in 4 out the 20 treated animals The incidence of the cutaneous reactions was therefore low (20%) and according to the criteria laid down in CLP (Reg 1272/2008/EC) and directive 67/548/EEC, the substance should not be considered as a skin sensitizer.

Executive summary:

The potential of the substance to induce delayed contact hypersensitivity was assessed in guinea pigs according to the OECD (n°406, 17th july 1992) and Commission Regulation (EC) (n°96/54/E.E.C., B.6, 30 july 1996) guidelines. The study was performed in compliance with the principle of Good Laboratory Practices regulations.

Thirty guinea pigs were allocated to two groups: a control group of five males and five females and a treated group of ten males and ten females. The induction phase was realized both by intradermal route on day 1 (Test material 0.1 % w/w in 0.9% NaCl) and by cutaneous route on day 8 (Test material 1% w/w in 0.9% NaCl). The challenge phase was realized on day 22 by cutaneous application of the test material at 1% w/w in 0.9% NaCl. The cutaneous reactions were scored 24 and 48 after the challenge phase.

No clinical signs and no deaths related to treatment were noted during the study. After the challenge application, no cutaneous reactions were observed in the animals of the control group. In the treated group, at the 24-hour reading, a discrete or moderate erythema was noted in 3/20 and 1/20 animals, respectively. It persisted at the 48-hour reading in 3/20 animals and appeared in 1/20 animals. Dryness of the skin was observed in 4/20 animals at the 48-hour reading. The persistent cutaneous reactions recorded in the treated group could be of sensitizing origin. However, as their incidence was low, the test item was not considered as a skin sensitizer.