Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Accepted, well-documented study report, which meets basic scientific principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Mouse ear swelling test (in vivo)
GLP compliance:
no
Type of study:
mouse ear swelling test
Justification for non-LLNA method:
The mouse ear swelling test (in vivo) was performed before the LLNA was set as preferred test method.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): C-1043 (Triethylamine).
- Molecular formula (if other than submission substance): C6H15N
- Molecular weight (if other than submission substance): 101.19
- Smiles notation (if other than submission substance): CCN (CC)CC
- Substance type: organic
- Physical state: clear liquid
- Storage condition of test material: at room temperature
- Other: Data of Receipt: August 21, 1986, received from Celanese Corporation

In vivo test system

Test animals

Species:
mouse
Strain:
other: Albino, CFL (BR)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories,Inc . Kingston, New York
- Age at study initiation: 6-8 weeks
- Weight at study initiation: 20-24 g
- Housing: group housed 2-5/cage (stainless steel with wiremesh bottoms).
- Diet (e.g. ad libitum): ad libitum. Purina Laboratory Chow, #5001,
- Water (e.g. ad libitum): ad libitum. Automatic watering system. Municipal water supply (Elizabethtown Water Co.)
- Acclimation period: 14 days
Currently acceptable practices of good animal husbandry were followed, e .g., Guide for the Care and Use of Laboratory Animals ; NIH Publication No. 85-23, Revised 1985.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 67-76 °F is considered an acceptable temperature range for mice; room temperature was monitored and recorded twice daily and mainained within this range to the maximum extent possible.
- Humidity (%): 30-70 % is considered an acceptable humidity range for mice; room humidity was monitored and recorded twice daily and maintained within this range to the maximum extent possible.
- Air changes (per hr): not reported.
- Photoperiod (hrs dark / hrs light): 12/12 (controlled by an automatic timer).

IN-LIFE DATES: From: To:

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
other: topical application to the clipped abdomen
Vehicle:
other: 70% ethanol
Concentration / amount:
0.1 mL was added to 9 .9 mL of 70% ethanol to produce a 0 .01 mL/mL (1 .0 % v/v} mixture.
Challengeopen allclose all
Route:
other: topical application to the dorsal and ventral surface of the ear
Vehicle:
other: 70% ethanol
Concentration / amount:
0.1 mL was added to 9 .9 mL of 70% ethanol to produce a 0 .01 mL/mL (1 .0 % v/v} mixture.
No. of animals per dose:
Range finding: 10
Sensitization study: Test animals: 20 (10 per group); Irritation Control animals: 15 (5 per group).
Details on study design:
RANGE FINDING TESTS: 10 mice (2 per concentration) were subjected to preliminary studies as follows:
Initially, 5%, 1%, 0 .5% and 0.1% concentrations were administered on the abdomen and 50 %, 25 %, 10 % and 5 % concentrations were administered on the ears. Based on the results, 100 % concentration was administered on the ears of a second set of animals. Each animal was dosed with only one concentration, on the abdomen and/or ear, dosing 2 animals per concentration (for a total of ten animals).
Day0: 100 µL of appropriate concentration was applied to the abdomen. In addition, 10 µL of the appropriate concentration was applied to both the dorsal and ventral surface of the left ear.
Days 1, 2 and 3: Animals were tape-stripped on the abdomen for five consecutive times and 100 µL of the appropriate concentration was applied to the abdomen.

Evaluation of responses:
1. Ear thickness on Day 1: The ear thickness of both left and right ear was measured 18-24 hours prior to and 24 hours after test material application.
2. Dermal responses on Day 4: The abdomen was evaluated 24 hours after the last exposure for the presence or absence of irritation (irritation was considered present if there was apparent erythema). Tissue destruction (subepidermal necrosis or eschar), was noted if present. [Note : Superficial necrosis or foci or small patches of necrosis is not indicative of permanent iissue destruction, i .e . corrosion].
Results: All concentrations of test material did not produce ear thikness. 5 % and 1 % of test material applied to abdomen produced subepidermal necrosis with eschar and edema, 0.5% produced subepidermal necrosis with eschar in one animal and small patches of subepidermal necrosis in another animal. 0.1 % produced small patches of subepidermal necrosis in one animal and focus of subepidermal necrosis.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 4 (days:0, 1, 2, and 3)
- Exposure period: 4 days
- Test groups: 10 animals
- Control group: 5 animals
- Site: clipped abdomen
- Frequency of applications: once each day
- Duration:
- Concentrations: 100 µL of whether 1 % TEA in 70 % ethanol or vehicle

B. CHALLENGE EXPOSURE
- No. of exposures:
- Day(s) of challenge: Day 10 Challenge, Day 17 Re-challenge
- Exposure period: on day 10 and day 17
- Test groups: 10 µL of the appropriate material was administered to both the dorsal and ventral surface of the left ear of each animal. 10 µL of the appropriate vehicle was administered to both the dorsal and ventrai surfaces of the right ear of the animals in the positive control group.
- Control group: yes
- Site: dorsal and venral surfaces of the left and right ear
- Concentrations: 100% TEA
- Evaluation (hr after challenge): Day 11 and 12 (24 and 48 hours after challenge) and Day 18 and 19 (24 and 48 hours after re-challenge)

OTHER:
Challenge controls:
In order to differentiate reactions produced by irritation from those produced by sensitization, five irritation control animals (previousiy treated w ith vehicle and FCA) were subjected to the same challenge procedure as the test animals.
Positive control substance(s):
yes
Remarks:
2,4-dinitrochlorobenzene (DNCB)

Results and discussion

Positive control results:
80 % of the animals treated with DNCB exhibited a positive sensitization response (test ear at least 20 % thicker than control ear), at 24-hours and 100% exhibited a positive response at 48 hours.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100 % TEA
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no ear swelling
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100% TEA. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no ear swelling.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100 % TEA
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no ear swelling
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100% TEA. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no ear swelling.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100 % TEA
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no irritation
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100% TEA. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no irritation.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 % TEA
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no irritation
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1005 TEA. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no irritation.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
1 % DNCB
No. with + reactions:
4
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
1 % DNCB
No. with + reactions:
5
Total no. in group:
5

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, C-1043 (TEA) exhibited no potential to produce sensitiiation in mice.
Executive summary:

The Mouse Ear Swelling Test was conducted to evaluate the allergic contact sensitization potential of TEA in mice. Albino Mice of CFL (BR) strain were used in all experiments. The concentrations of test material for induction and challenge phases were determined in a range-finding study. For induction, 1% TEA in 70% ethanol was applied to abdomens of animals. For the challenge, 100% TEA was applied topical, to the dorsal and ventral surface of the ear of the animals. 2,4- dinitrochlorobenzene (DNCB) was used as positive control. Freund's Complete Adjuvant (FCA) consisted from a mixture of paraffin oil and an emulsifier with mycobacteria. On the day 0 (Induction phase), a 50% concentration of FCA in water was administered by injection to two clipped and stripped of horny layer sites of abdomen, one on the left side and one on the right side. 100 µL of whether test material, positive control or appropriate vehicle was administered directly to the test site, on the abdomen of the animals. This procedure was repeated on day 1, 2, and 3 (Induction phase). On the day 9 before the challenge administration, both ears of each animals were examined for the presence of irritation (erythema, edema). No pre-existing irritation was seen in any animal, therefore, all animals remained on study. On the day 10, test animals were challenged with whether 10 µL of 100% TEA or DNCB on the dorsal and ventral surface of the left ear of each animal. 10 µL of appropriate vehicle was administered in the same manner on the right ear. In order to differentiate reactions produced by irritation from those produced by sensitization, five irritation control animals (previousiy treated with vehicle and FCA) were subjected to the same challenge procedure as the test animals. On the day 16, before pre-challenge administration, the ears were examined again for the presence of irritation. At re-challenge, the same procedures were followed as were followed at challenge (Day 10), except the ears were switched, i.e., only the right ear was dosed with the test material; the left ear was not treated. Irritation control animals, previously treated with vehicle and FCA only and not treated at challenge) were subjected to the same challenge procedure as the test animals. The first and the second readings (24 and 48 hours, respectively) were performed after challenge and re-challenge application.

Except for one animal which did not recover from the anesthesia at the 24-hour Challenge Response Measurement (DNCB irritation control animal , Group 1B), all animals survived throughout the study. 80 % of the animals treated with DNCB exhibited a positive sensitization response (test ear at least 20 % thicker than control ear), at 24-hours and 100 % exhibited a positive response at 48 hours. There were no positive responders in the irritation control group. This positive response to a known sensitizer demonstrated the susceptibility of this group of animals to sensitization. Animals challenged with TEA exhibited no positive responses at challenge or rechallenge in either the test group or irritation control groups.