Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2004-03-22 to 2004-10-22
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP study performed according to OECD guideline N° 406. No detail on the test animal and no data on the positive control.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
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:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
: no detail on the test animal and no data on the positive control.
Principles of method if other than guideline:
Not applicable
GLP compliance:
no
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: compact

In vivo test system

Test animals

Species:
guinea pig
Strain:
not specified
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: no data (young healthy adult animals)
- Weight at study initiation: no data
- Housing: no data
- Diet (e.g. ad libitum): specific diet for guinea pigs
- Water (e.g. ad libitum): drinking water, no more detail.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C (+/-3°C)
- Humidity (%): 30-70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light

IN-LIFE DATES: no data

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: deionized water
Concentration / amount:
- Intradermal induction: 12% in dieonized water
-Topical induction: 100% in dieonized water (0.5mL)
-Topical challenge: 100% in dieonized water (0.5mL)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: deionized water
Concentration / amount:
- Intradermal induction: 12% in dieonized water
-Topical induction: 100% in dieonized water (0.5mL)
-Topical challenge: 100% in dieonized water (0.5mL)
No. of animals per dose:
20 animals for the test group
10 animals for the control group
Details on study design:
RANGE FINDING TESTS:
One animal was exposed by intradermal injection to the test item at 100, 75 and 50% of concentration, and a second animal received 25, 12 and 6% -for each concentration, one pair of injections was made, one of each pair lies on each side of the midline. Evaluations were done 24 hours after injections.
Three animals were exposed by topical application to the test item. One animal received the test substance at 100% and the two others at 50 and 25%. Evaluations were done 48 hours after topical applications.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 0 (injection) and day 7 (topical)
- SLS application: yes (10% in vaseline)
- Test groups: test substance (12% in deionized water) + test substance 12% in FCA
- Control group: deionized water in FCA
- Site: for each concentration, one pair of injections was made, one of each pair lies on each side of the midline
- Frequency of applications: the topical application was performed one week following the intradermal injection. Single exposure in each case
- Duration: Day 0 to Day 20 (with a rest period from Day 9 to Day 20)
- Concentrations: 12% (injection), 100% (topical)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 21
- Exposure period: 24 hours
- Test groups and control group: topical application of the test substance in deionized water (one flank) and deionized water alone (other flank)
- Site: right flank (test material) and left flank (vehicle)
- Concentrations: 100% (wetted with deionized water, 0.5 mL)
- Evaluation (hr after challenge): 24 and 48 hours after removal of the dressing

OTHER:
Magnusson and Kligman grading score for the evaluation of challenge:
* No visible change..........0
*Discrete or patchy erythema.........1
*Moderate and confluent erythema......2
*Intense erythema and swelling.............3
Challenge controls:
No data
Positive control substance(s):
not specified

Results and discussion

Positive control results:
No data

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no data
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no data.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no data
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no data.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
-
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no data
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: -. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no data.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
-
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no data
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: -. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no data.

Any other information on results incl. tables

Table 7.4.1/1:Results of the preliminary intradermal injection with two animals.

Animal number

Test item concentration

Oedema/Erythema score (24 hours)

01

100%

3

75%

3

50%

2

02

25%

1

12

1

6%

0

 

Table 7.4.1/2:Results of the preliminary topical application with three animals.

Animal number

Test item concentration

Oedema/Erythema score (48 hours)

01

100%

0

02

50%

0

03

25%

0

Table 7.4.1/3: Reaction at challenge at 24 h and 48 h in negative control group and test group (100%)

Animal

number

Reaction at challenge

24 h

48 h

Negative Control Group

01

0

0

02

0

0

03

0

0

04

0

0

05

0

0

06

0

0

07

0

0

08

0

0

09

0

0

10

0

0

Test Group (100%)

01

0

0

02

0

0

03

0

0

04

0

0

05

0

0

06

0

0

07

0

0

08

0

0

09

0

0

10

0

0

11

0

0

12

0

0

13

0

0

14

0

0

15

0

0

16

0

0

17

0

0

18

0

0

19

0

0

20

0

0

 

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this test, ORTHO-DIETHOXY BENZENE (ODEB) does not induce delayed contact hypersensitivity in guinea pigs. The test substance was therefore considered as not sensitizing and not classified according to the CLP Regulation (EC) N°1272/2008 and of the Directive 67/548/EEC criteria.
Executive summary:

In a non- GLP skin sensitisation study (Siqueira, 2004), male guinea pigs were used to test the skin contact sensitisation potential of ortho-Diethoxybenzene (ODEB). The method of Guineapig maximisation test was used (OECD 406).

In a preliminary study, the skin irritation potential of the test item was assessed in two animals after an intradermal injection (6, 12, 25, 50, 75 and 100%) and on three animals following a topical application (25, 50 and 100%). Based on the results of this pilot study, the concentration of the test substance used in the induction phase, intradermal and topical applications were, respectively, 12% in deionized water and 100% for the main study, and in the challenge phase the tested concentration was 100%.

In the main study performed on 20 animals for the test group and 10 animals for the control group, three test sites were tested in each animal. In the first induction stage intradermal injections of

the test item (12% in deionized water) +/- Freund Complete Adjuvant or with the vehicle were performed on each animal. One week later, the animals were pre-treated with sodium lauryl sulphate (to promote a skin irritation) and then topically exposed to the test item at 100% (wetted with deionized water) or the vehicle. Two weeks following this second induction stage, animals were challenged by a topical application of the test item at 100% (wetted with deionized water) for 24 hours. At 24 and 48 hours after the challenge phase at concentrations of 100%, it was not observed any skin reaction in the animals receiving the test substance or deionized water. As all tested animals (20) didn’t show hypersensivity with the ODEB, no classification is warranted according to the CLP Regulation (EC) N°1272/2008 and of the Directive 67/548/EEC criteria.

 

This skin sensitisation study is classified as acceptable. It satisfies the guideline requirement for a skin sensitisation in the guinea pig.