Registration Dossier

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:
27 Apr - 17 Jun 1994
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study with acceptable restrictions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
no reliabilty check included
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
yes
Remarks:
no reliabilty check included
GLP compliance:
yes
Type of study:
Buehler test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): only trade name given
- Chemical denomination: Fatty acid, C16-18, 1,2-Ethandiyl Bis
- Substance type: pure substance
- Physical state: yellowish solid
- Analytical purity: 100%
- Lot/batch No.: 62393218
- Expiration date of the lot/batch: December 31, 1994
- Storage condition of test material: dark and room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Hsd/Win:DH
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Age at study initiation: approx. 6 weeks
- Weight at study initiation: 340 g
- Housing: Groups of 2-3 animals in Makrolon Type IV cages with standard softwood bedding. Change of bedding: two times a week.
- Diet: Pelleted Altromin Maintenance Diet 3022 (Altromin GmbH, Lage, Germany), ad libitum
- Water: Tap water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 45-75
- Air changes (per hr): at least 8
- Photoperiod (hrs dark / hrs light): 12/12
- Other: Music during light hours

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
peanut oil
Concentration / amount:
Induction: 25%
Challenge: 20%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
peanut oil
Concentration / amount:
Induction: 25%
Challenge: 20%
No. of animals per dose:
3 (preliminary), 5 (dose finding), 10 (controls), 20 (in test groups)
Details on study design:
PRELIMINARY STUDY
Preliminary tests were carried out to determine suitable concentrations for induction and challenge. The concentrations in a dose of 0.5 g (50%) and 0.5 mL (25%, 12.5% and 6%) were tested. The concentrations were applied on the left flanks of three animas in succession. The exposure was terminated after 6 h by removing the plaster and cleaning the skin with propylene glycol 20%.

DOSE FINDING
The non-irritating concentrations for the challenge were checked in five animals of the control group (treated with the vehicle one week before the 1st challenge). The concentrations applied on the right flank in a dose of 0.5 mL were 10%, 15%, 20% and 25%. The exposure was terminated after 6 h by removing the plaster and cleaning the skin with propylene glycol 20%. 24 and 48 hours later, dermal effects were evaluated.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 6 h
- Test groups: test substance in peanut oil
- Control group: peanut oil
- Site: left flank
- Frequency of applications: every 7 days
- Duration: Days 0-21
- Concentrations: 25%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 35
- Exposure period: 6 h
- Test groups: test substance in peanut oil
- Control group: test substance in peanut oil
- Site: bilaterally to both sheared flanks (caudal) of animals of the treatment and the control group
- Concentrations: 20%
- Evaluation (hr after challenge): 24, 48 and 72 h after patch removal

EVALUATION CRITERIA
- see Table 1
Challenge controls:
The control group is actually a challenge control
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
20%
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
slight dermal effects at the left or right flank
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 20%. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: slight dermal effects at the left or right flank.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
20%
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
slight dermal effects at the left flank
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 20%. No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: slight dermal effects at the left flank.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
20%
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
slight dermal effects at the left or right flank
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 20%. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: slight dermal effects at the left or right flank.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
20%
No. with + reactions:
9
Total no. in group:
20
Clinical observations:
slight dermal effects in 8 animals and weak dermal effects in one animal at the left and/or right flank
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 20%. No with. + reactions: 9.0. Total no. in groups: 20.0. Clinical observations: slight dermal effects in 8 animals and weak dermal effects in one animal at the left and/or right flank.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
20%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
20%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading:

Any other information on results incl. tables

PRELIMINARY STUDY

In the preliminary study no reaction of the test substance was observed up to 50% in the test animals. According to the author a solution of 25% was considered the minimal irritating concentration. The concentration chosen for the induction period was 25%.

DOSE FINDING

The tested solutions did not provoke any skin reactions at the 5 dose finding animals. The concentration chosen for challenge was 20%.

MAIN STUDY

Induction readings:

1 hour after the 3rd induction, three animals of the control group (15 animals: 10 control animals plus 5 animals out of the dose finding group) and four animals of the treatment group showed weak skin reactions. Only for one animal out of the treatment group slight skin reactions were observed after 24 hours (see Table 2).

Table 2. Skin reactions 1 and 24 hours (left flank) after 3rd induction.

Skin reaction/ Value

1 hour

c (15) / t (20)

24 hours

c (15) / t (20)

none / 0

12

16

15

19

slight / 0+

0

0

0

1

weak / 1

3

4

0

0

moderate / 2

0

0

0

0

strong / 3

0

0

0

0

c: control group; t: treatment group; ( ): number of animals

Challenge readings:

24 hours after challenge application, 2/20 animals of the treatment group (10%) and 2/10 animals of the control group (20%) showed slight dermal effects (see Table 3 and Table 4). After 48 hours, 8/20 animals of the treatment group (40%) and 2/10 animals of the control group (20%) showed slight skin reactions. Only for one treated animal (5%) weak skin reactions were observed after 48 hours. After 72 hours no skin reactions were observed in any animal.

Table 3. Summary of positive animals in % after challenge.

Skin reaction/ Value

control group

l / r

treatment group

l / r

 

24 h

48 h

72 h

24 h

48 h

72 h

slight / 0+

20

20

0

10

40

0

weak / 1

0

0

0

0

5

0

l: left flank; r: right flank; h: hours

Table 4. Individual gradings for animals of the treatment and the control groups.

 

Flank

Animal number

1st challenge

                            left                                                          right

     24 h             48 h             72 h            24 h           48 h             72 h

t1

0

0+

0

0

0+

0

t2

0

0+

0

0

0

0

t3

0

1

0

0

1

0

t4

0

0

0

0

0

0

t5

0

0

0

0

0+

0

t6

0

0

0

0

0

0

t7

0+

0+

0

0

0

0

t8

0

0+

0

0

0+

0

t9

0

0

0

0

0

0

t10

0

0

0

0

0

0

t11

0

0

0

0

0

0

t12

0

0

0

0

0

0

t13

0

0

0

0

0+

0

t14

0+

0+

0

0

0+

0

t15

0

0

0

0

0+

0

t16

0

0

0

0

0

0

t17

0

0

0

0

0

0

t18

0

0

0

0

0

0

t19

0

0

0

0

0

0

t20

0

0

0

0

0

0

c1

0

0

0

0

0

0

c2

0

0

0

0

0

0

c3

0

0

0

0

0

0

c4

0

0

0

0

0

0

c5

0

0

0

0

0

0

c6

0

0+

0

0+

0+

0

c7

0

0

0

0

0

0

c8

0+

0+

0

0

0

0

c9

0

0

0

0

0

0

c10

0

0

0

0

0

0

c: control group; t: treatment group; h: hours

grading: none / 0; slight / 0 +; weak / 1

Mortality/Body weight

No mortality was observed during the test period. No significant difference in the gain of the body weight between the treatment and the control group was observed.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified