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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August 14, 1995 to December 22, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
other: EEC Council directive 67/548/EEC, September 19, 1984
Deviations:
no
Qualifier:
according to
Guideline:
EPA OTS 798.4100 (Skin Sensitisation)
Deviations:
no
Principles of method if other than guideline:
no devaitions which would compromise the validity of the study.
GLP compliance:
yes
Remarks:
EPA TSCA GLP guidelines (40 CFR Part 792) and OECD GLP's "OECD Principles of Good Laboratory Practice, C(81)30(Final)Annex 2. May 12, 1981.
Type of study:
Buehler test
Justification for non-LLNA method:
The Buehler test (in vivo) was performed before the LLNA was set as preferred test method.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid
Details on test material:
BUTYLDIETHANOLAMINE 1029- Name of test material (as cited in study report):
- Substance type: alkyl alkanol amine
- Physical state: Water white to yellow liquid
- Analytical purity: 99.7%
- Purity test date: 1995-08-03
- Lot/batch No.: E-08-F
- Expiration date of the lot/batch: September 3, 1995
- Stability under test conditions:
The Sponsor assumes responsibility for purity and stability determinations (including under test conditions). Information on composition and method of synthesis was held by the Sponsor. Analyses of test material/vehicle for concentration, solubility, homogeneity, and stability were not done by Hill Top Biolabs.
- Storage condition of test material: Room temperature
- Other: product code: 01029

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: from a U.S.D.A. licensed supplier (Animal Supplier: Harlan Sprague Dawley, Inc. P.O. Box 29176 Indianapolis, Indiana 46229)
- Age at study initiation: young adult
- Weight at study initiation: 342 to 467 grams
- Housing: individually in wire mesh suspension cages
- Diet (e.g. ad libitum): ad libitum (Teklad Guinea Pig Diet)
- Water (e.g. ad libitum): ad libitum (tap water)
- Acclimation period: at least 4 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 29, 1995 To: October 16, 1995

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Remarks:
Undiluted distilled
Concentration / amount:
- Pilot test: undiluted, 50%, 25%, 10%, 5%, 2.5%, 1.0%, and 0.5%;
- Main test: 50% (challenge);
- Main test: 50% and 1% (re-challenge).
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Remarks:
Undiluted distilled
Concentration / amount:
- Pilot test: undiluted, 50%, 25%, 10%, 5%, 2.5%, 1.0%, and 0.5%;
- Main test: 50% (challenge);
- Main test: 50% and 1% (re-challenge).
No. of animals per dose:
- Pilot test: 8 animals (two groups of four animals each);
- Main test: 20 (test animals); 10 (vehicle control animals)
Details on study design:
RANGE FINDING TESTS:
Irritation Screening (Pilots):
The irritation phase had the purpose of determining the proper level of test material to be used in the induction and challenge phase. The irritation potential of Butyldiethanolamine 1029 at levels of undiluted, 50%, 25%, 10%, 5%, 2.5%, 1.0%, and 0.5% was evaluated in two groups of four animals each. Four levels of test material were evaluated per animal, such that each animal in a given pilot group was exposed to the same levels. Dilutions of the test material were prepared w/v in distilled water. The position of the different concentrations of the test material on the animals was varied to adjust for possible site-to-site variation in response (See diagram below for available skin sites, attached to this file).
The day prior to test material exposure, the hair was removed from each of the animal's backs using a small animal clipper. Closed patches were applied to the animals in the following manner: A 0.3 ml amount of each test preparation was applied into a 25 mm Hill Top Chamber® (This 0.3 ml volume is reduced from the 0.5 ml volume referenced in the EEC Guidelines due to the volume limit of the 25 mm Hill Top Chamber®). The animal was placed into the restrainer and the chamber(s) were applied to the clipped surface as quickly as possible. The chamber(s) were occluded with rubber dental dam pulled taut and fastened to the bottom of the restrainer with clips. The restrainer was adjusted to minimize movement of the animal during exposure. Approximately six hours later (± 15 minutes), the dental dam and chamber(s) were removed, the animal was taken from the restrainer, and placed in its cage. The day following the irritation exposure all animals were depilated and scored.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 6 hours
- Test and control groups:
The left shoulder (Site 1) of each animal was clipped with a small animal clipper the day before exposure (See diagram below for skin site, attached to this file). The clipped areas were exposed to the respective test or vehicle material and the animals were restrained as previously described under "Irritation Screening". The procedure was repeated at the same site once every seven days for the next fourteen days for a total of three approximate six-hour exposures (± 15 minutes). (See Protocol Deviations section Page 20 of report). Induction sites were graded approximately 24 hours (± 15 minutes) after completion of each induction exposure using the grading scale described under "Observations". After the last induction exposure, the animals were left untreated for fourteen days before primary challenge.
- Site: 1(left shoulder)
- Frequency of applications: once every seven days
- Duration: 14 days
- Concentrations: undiluted

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: fourteen days after the last induction exposure.
- Exposure period: 6 hours
- Test and control groups: The same exposure procedure was used as for the "Induction Phase", but the chambers were applied to the test animals and vehicle animals at the lower left flank (Site 2).The patch sites were varied to adjust for possible site-to-site variation in response. (See diagram below for available skin sites, attached to this file).
- Site: 2 (lower left flank)
- Concentrations: 50 % (w/v in distilled water)
- Evaluation (hr after challenge): 24 and 48

OTHER: RE-CHALLENGE (see in "Any other information on materials and methods").
Challenge controls:
The vehicle control animals were common to this study and studies 95-8736-21 and 95-8737-21; at the challenge, they received one patch of each test material using Sites 2, 4, and 5, respectively.
Positive control substance(s):
yes
Remarks:
routinely analysed at the test facility

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight but confluent, or moderate patchy erythema (score of 1).
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50%. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight but confluent, or moderate patchy erythema (score of 1)..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50 %
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight but confluent, or moderate patchy erythema (score of 1)
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight but confluent, or moderate patchy erythema (score of 1).
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no positive responders with score of 1.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no positive responders with score of 1.
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
50 %
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight but confluent, or moderate patchy erythema (score of 1)
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight but confluent, or moderate patchy erythema (score of 1).
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
50 %
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight but confluent, or moderate patchy erythema (score of 1)
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight but confluent, or moderate patchy erythema (score of 1).
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no positive responders with score of 1.
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 1 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no positive responders with score of 1..
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 1 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no positive responders with score of 1.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no positive responders with score of 1.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no positive responders with score of 1.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 1 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no positive responders with score of 1.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive responders with score of 1
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 1 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no positive responders with score of 1.
Group:
positive control
Remarks on result:
other: routinely analysed at the test facility

Any other information on results incl. tables

Results of the primary irritation, induction, primary challenge, and rechallenge applications are presented in Tables 1 through 7, respectively (please see attached to this file). Individual body weight data are presented in Table 8.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the criteria of significant skin sensitizing effects outlined in the Guidance on the Application of Regulation (EC) No. 1272/2008 (2013), a substance should be considered as sensitizing in the Buehler occluded patch test if redness is observed in >= 15 % of the test animals. Butyldiethanolamine as 50 % dilution induced a positive response (score of 1 = slight but confluent, or moderate patchy erythema) at both reading time points in one animal at challenge and in another one animal at re-challenge. However, 1 out of 20 animals amounts to 5 %. Moreover, no positive responders were observed at rechallenge with 1 % of butyldiethanolamine. Therefore, Butyldiethanolamine did not induce a significant skin sensitisation effect and does not need to be classified and labelled as skin sensitiser in accordance with Regulation (EC) No. 1272/2008.
Executive summary:

The test material, Butyldiethanolamine 1029, was evaluated for primary irritation at levels of undiluted, 50%, 25%, 10%, 5%, 2.5%, 1%, and 0.5%. All formulations were prepared w/v in distilled jwater. The 50% formulation formed a clear solution with a yellow tint which decreased in color to clear colorless solutions as the concentrations decreased. Undiluted Butyldiethanolamine 1029 produced grades of 1, 50%, 25%, and 5% produced grades of ±, while 10%, 2.5%, 1%, and 0.5% produced grades of ± and 0. An undiluted concentration of Butyldiethanolamine 1029 was chosen for use at induction for the test group. This level was chosen as the highest concentration causing no greater than mild to moderate primary irritation. Undiluted distilled water was utilized at induction for the vehicle control group. A 50% w/v concentration of Butyldiethanolamine 1029 in distilled water was chosen for use at primary challenge for the test group and the vehicle control group. This level was chosen as the highest concentration causing no more than slight primary irritation. The incidence and severity of responses were calculated as follows:

Group

Materiala

CONCbc

Incidence of Responses

Mean Severity Scores

24-Hours

48-Hours

0

±

1

2

3

0

±

1

2

3

24-hr

48-hr

Primary Challenge

Test

TMID

50%

2

17

1

0

0

8

11

1

0

0

0.5

0.3

Vehicle Control

TMID

50%

2

7

0

0

0

7

2

0

0

0

0.4

0.1

Re-challenge

Test

TMID

50%

7

12

1

0

0

6

13

1

0

0

0.4

0.4

TMID

1%

12

8

0

0

0

15

5

0

0

0

0.2

0.1

Vehicle Control

TMID

50%

3

6

0

0

0

7

2

0

0

0

0.3

0.1

TMID

1%

6

3

0

0

0

7

2

0

0

0

0.2

0.1

aTMID - BUTYLDIETHANOLAMINE 1029.

bCONC = Concentration.

cTest material formulated w/v in distilled water.

A single test animal (T-13) had a grade 1 response at challenge and another test animal (T-20) responded at rechallenge to the 50% dilution of test material but not to 1%. Both responses are suggestive of sensitization. No control animal responded with a grade 1, suggesting that sensitization had occurred.