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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:
1994-10-11 - 1995-02-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Scientifically valid GLP study on the registered substance itself according to US EPA guideline 81-6.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EPA OPP 81-6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
Sufficient data from Buehler test conducted in 1994

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction Products of Diphosphorus Pentaoxide with n-Alcohols, C8-10 (even), salted with Amines, C12-14, Tert-alkyl
EC Number:
943-016-1
Molecular formula:
Too complex
IUPAC Name:
Reaction Products of Diphosphorus Pentaoxide with n-Alcohols, C8-10 (even), salted with Amines, C12-14, Tert-alkyl
Test material form:
other: liquid
Details on test material:
- Substance type: pure substance
- Storage condition of test material: Room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Hartley-derived albino
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc., Haslett, Michigan
- Age at study initiation: young adult
- Weight at study initiation: 300-500g
- Housing: individually in suspended stainless steel cages
- Diet (e.g. ad libitum): Purina Certified Guinea Pig Chow #5026, ad libitum
- Water (e.g. ad libitum): Municipal tap water treated by reverse osmosis or deionization (back-up system), ad libitum
- Acclimation period: min. 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 58-74°F (14.4-23.3°C)
- Humidity (%): 34-68%
- Air changes (per hr): 10-12
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: USP grade mineral oil
Concentration / amount:
25%, 50%, 75%, 100% (Topical Range-Finding Study)
50% (Induction)
1.0%, 2.5%, 5.0%, 10.0% (Second Topical Range-Finding Study)
10% (Challenge)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: USP grade mineral oil
Concentration / amount:
25%, 50%, 75%, 100% (Topical Range-Finding Study)
50% (Induction)
1.0%, 2.5%, 5.0%, 10.0% (Second Topical Range-Finding Study)
10% (Challenge)
No. of animals per dose:
each 2 m/f (First & Second Topical Range-Finding Study)
each 10 m/f (Induction & Challenge, Main Test)
each 5 m/f (Challenge Control, Main Test)
each 3 m/f (Induction & Challenge, DNCB, Main Test)
each 2 m/f (DNCB control)
Details on study design:
RANGE FINDING TESTS:
Dosing: On the day prior to dose administration, four topical range-finding guinea pigs were weighed and the hair removed from the right and left side of the animals with a small animal clipper. Care was taken to avoid abrading the skin during clipping procedures. On the following day, four concentrations of the test article were prepared and each concentration (0.3ml) was applied to each topical range-finding animal in a 25 mm Hilltop chamber. Following chamber application, the trunk of the animal was wrapped with elastic wrap which was secured with adhesive tape to prevent removal of the chamber and the animal was returned to its cage. Approximately six hours after chamber application, the elastic wrap, tape and chambers were removed. The test sites were then wiped with gauze moistened in USP grade mineral oil followed by dry gauze to remove test article residue.
Dermal Observations: The test sites of the topical range-finding animals were graded for irritation at approximately 24 and 48 hours following chamber application.

MAIN STUDY
A. INDUCTION EXPOSURE (procedure see above)
- No. of exposures: 3
- Exposure period: 3x6 h
- Test groups: each 10 m/f, 0.3ml of a 50% solution of the test item
- Control group: each 3 m/f, 0.3ml of a 0.5% solution of DNCB
- Site: left side of the trunk
- Frequency of applications: on day 0, 7, and 14
- Duration: 6h
- Concentrations: 50% (test item), 0.5% (DNCB)

B. CHALLENGE EXPOSURE (procedure see above)
- No. of exposures: 1
- Day(s) of challenge: day 28
- Exposure period: 6h
- Test groups: each 10 m/f, 0.3ml of a 10% solution of the test item
- Control group: each 5 m/f for challenge control, each 3 m/f for DNCB, each 2 m/f for DNCB control
- Site: right side
- Concentrations: 10% (test item), 0.1% / 0.2% (DNCB)
- Evaluation (hr after challenge): 24h & 48h
Challenge controls:
each 5 m/f animals, 0.3ml of a 10% solution of the test item
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene (DNCB)

Results and discussion

Positive control results:
Following challenge with DNCB, 6/6 DNCB test animals were noted to have a substantially stronger dermal response than was observed in the corresponding DNCB control animals. Group mean dermal scores were also noted to be higher In the DNCB test animals as compared to those of the DNCB control animals.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10%
No. with + reactions:
15
Total no. in group:
20
Clinical observations:
Slight patchy erythema; mean dermal score: 0.4
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10%. No with. + reactions: 15.0. Total no. in groups: 20.0. Clinical observations: Slight patchy erythema; mean dermal score: 0.4.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10%
No. with + reactions:
9
Total no. in group:
20
Clinical observations:
Slight patchy erythema; mean dermal score: 0.2
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10%. No with. + reactions: 9.0. Total no. in groups: 20.0. Clinical observations: Slight patchy erythema; mean dermal score: 0.2.
Reading:
1st reading
Hours after challenge:
24
Group:
other: Challenge control
Dose level:
10%
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
Slight patchy erythema; mean dermal score: 0.4
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: Challenge Control. Dose level: 10%. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: Slight patchy erythema; mean dermal score: 0.4.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Challenge control
Dose level:
10%
No. with + reactions:
4
Total no. in group:
10
Clinical observations:
Slight patchy erythema; mean dermal score: 0.2
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: Challenge Control. Dose level: 10%. No with. + reactions: 4.0. Total no. in groups: 10.0. Clinical observations: Slight patchy erythema; mean dermal score: 0.2.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.1%, 0.2%
No. with + reactions:
6
Total no. in group:
6
Clinical observations:
Severe erythema with or without edema; mean dermal score: 3.0
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.1%, 0.2%. No with. + reactions: 6.0. Total no. in groups: 6.0. Clinical observations: Severe erythema with or without edema; mean dermal score: 3.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.1%, 0.2%
No. with + reactions:
6
Total no. in group:
6
Clinical observations:
Severe erythema with or without edema; mean dermal score: 3.0
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.1%, 0.2%. No with. + reactions: 6.0. Total no. in groups: 6.0. Clinical observations: Severe erythema with or without edema; mean dermal score: 3.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: DNCB control
Dose level:
0.1%, 0.2%
No. with + reactions:
4
Total no. in group:
4
Clinical observations:
Slight patchy erythema; mean dermal score: 0.5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: DNCB control. Dose level: 0.1%, 0.2%. No with. + reactions: 4.0. Total no. in groups: 4.0. Clinical observations: Slight patchy erythema; mean dermal score: 0.5.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: DNBC control
Dose level:
0.1%, 0.2%
No. with + reactions:
4
Total no. in group:
4
Clinical observations:
Slight patchy erythema; mean dermal score: 0.5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: DNBC control. Dose level: 0.1%, 0.2%. No with. + reactions: 4.0. Total no. in groups: 4.0. Clinical observations: Slight patchy erythema; mean dermal score: 0.5.

Any other information on results incl. tables

Table 8: Individual induction data – Test item

Animal No. / Sex

Induction 1 Dermal Scores

Induction 2 Dermal Scores

Induction 3 Dermal Scores

50%

50%

50%

24h

48h

24h

48h

24h

48h

3553/M

±

0

±

±

±

±

3554/M

±

±

±

±

±

±

3555/M

±

0

±

±

±

±

3556/M

±

±

±

±

±

±

3557/M

±

±

±

0

1ED-1

±

3558/M

±

±

±ED-1

±

±

±

3560/M

±

±

±

±

±

±

3561/M

±

0

±

0

±

±

3562/M

±

±

±

±

1ED-1

1

3563/M

±

±

±

±

1ED-1

1ED-1

3635/F

±ED-1

±

±

±

±

±

3636/F

±

±

±

±

±

±

3637/F

±

±

±

±

1ED-1

±

3638/F

±

±

1ED-1

1ED-1

1ED-1

1ED-1

3639/F

±

±

±

0

±

±

3640/F

±

0

1ED-1

1ED-1

1ED-1

1ED-1

3641/F

±

±

±

±

1ED-1

1ED-1

3642/F

±ED-1

±

±

±

1ED-1

±

3643/F

±ED-1

±ED-1

1ED-1

1ED-1

1ED-1

±

3644/F

±

±

±

±

1ED-1

1

See tables 4-6 for definition of codes

 

 

Table 9: Individual induction data – DNCB Test

Animal No. / Sex

Induction 1 Dermal Scores

Induction 2 Dermal Scores

Induction 3 Dermal Scores

0.5%

0.5%

0.5%

24h

48h

24h

48h

24h

48h

3579/M

±YST

±YST

3ED-3,ES-3,BLA-2,YST

3ED-2,ES-4,BLA-2,YST

3ED-3,ES-2,BLA-4,YST

3ED-2,ES-2,BLA-4,YST

3580/M

±YST

0YST

3ED-3,ES-3,BLA-2,YST

3ED-3,ES-4,YST

3ED-2, BLA-2,ES-1,YST

3ED-2, BLA-4,ES-2,YST

3581/M

±YST

±YST

3ES-2,BLA-4,ED-2,YST

3ES-2,BLA-2,ED-2,YST

3BLA-3, ED-2YST

3ES-1,ED-2,BLA-3,YST

3661/F

±YST

±YST

3ES-2,BLA-4,ED-3,YST

3ES-3,BLA-2,ED-4,YST

3BLA-3, ED-2YST

3BLA-3,ED-2,YST

3662/F

±YST

±YST

3ES-3,BLA-3,ED-3,YST

3ES-4,ED-3,BLA-2,YST

3ES-1,BLA-3,ED-2,YST

3ES-2,BLA-3,ED-2,YST

3663/F

±YST

±YST

3ES-2,BLA-4,ED-3,YST

3ES-3,BLA-2,ED-2,YST

3BLA-3, ED-2YST

3ES-1,BLA-3,ED-2,YST

See tables 4-6 for definition of codes

 

 

Table 10: Individual challenge data – Test item

        Group

Animal No. / Sex

Dermal Scores

10%

24h

48h

Test

3553/M

±

±

3554/M

±

±

3555/M

±

0

3556/M

±

±

3557/M

±

0

3558/M

0

0

3560/M

±

±

3561/M

0

0

3562/M

±

0

3563/M

±

0

3635/F

±

±

3636/F

0

0

3637/F

±

±

3638/F

±

±

3639/F

±

0

3640/F

±

0

3641/F

±

±

3642/F

0

0

3643/F

±

±

3644/F

0

0

Mean

0.4

0.2

Challenge Control

3564/M

±

±

3565/M

±

±

3566/M

±

0

3567/M

0

0

3568/M

±

±

3645/F

±

±

3646/F

0

0

3647/F

±

0

3648/F

±

0

3649/F

0

0

Mean

0.4

0.2

For the purpose of calculation, ± = 0.5

See tables 4-6 for definition of codes

 

 

Table 11: Individual challenge data – DNCB

Group

Animal No. / Sex

Dermal Scores

Dermal Scores

0.1%

0.2%

24h

48h

24h

48h

Test

3579/M

3ES-3,BLA-2,ED-3

3ES-4,BLA-1,ED-2

3BLA-4,ED-3

3ES-2,BLA-2,ED-2

3580/M

3ES-2,BLA-3,ED-2

3ES-3,BLA-2,ED-3

3ES-1,BLA-4,ED-2

3ES-1,BLA-4,ED-2

3581/M

3ES-2,BLA-4,ED-2

3ES-3,BLA-2,ED-3

3BLA-4,ED-3

3ES-1,BLA-4,ED-2

3661/F

3BLA-3,ED-2

3ES-3,BLA-2,ED-3

3BLA-4,ED-2

3ES-2,BLA-3,ED-2

3662/F

3ES-2,BLA-3,ED-2

3ES-3,BLA-2,ED-3

3ES-2,BLA-3,ED-2

3ES-3,BLA-2,ED-2

3663/F

3BLA-3,ED-2

3ES-3,BLA-2,ED-3

3BLA-4,ED-2

3ES-1,BLA-3,ED-2

Mean

3.0

3.0

3.0

3.0

Challenge Control

3564/M

±

±

±

±

3565/M

±

±

±

±

3645/F

±

±

±

±

3646/F

±

±

±

±

Mean

0.5

0.5

0.5

0.5

For the purpose of calculation, ± = 0.5

See tables 4-6 for definition of codes

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The study is well-documented and was performed according to EPA OPP 81-6 (Skin sensitization), under GLP conditions. Hence, the information provided in the report can be considered reliable. In consequence, the results are suitable for the assessment of the sensitizing potential of the test item. Under the conditions of this test, the test item was not considered to be a sensitizer in guinea pigs. The results of the DNCB positive control study demonstrated that a valid test was performed and indicated that the test system could detect potential contact sensitizers.
Executive summary:

In a dermal sensitization study according to EPA OPP 81-6 (Skin sensitization) with the test item in USP grade mineral oil, young adult Hartley-derived albino (10/sex in the main test) were tested using the modified Buehler assay. 1-chloro-2,4-dinitrobenzene (DNCB) served as positive control.

 

Regarding the Topical Range-Finding Studies, the results of the first range-finding study indicated that a test article concentration of 50% was considered appropriate for Induction. The results of the second range-finding study indicated that a test article concentration of 10% was considered appropriate for challenge.

 

In the Main Sensitization Study, following challenge with 10% w/v of the test item in USP grade mineral oil, the animal responses met the negative criteria for sensitization based on the evaluation criteria. Dermal reactions in all test and challenge control animals were limited to scores of 0 to ± (0% net responders). Group mean dermal scores were noted to be similar in the test animals as compared to the challenge control animals.

Following challenge with DNCB, 6/6 DNCB test animals were noted to have a substantially stronger dermal response than was observed in the corresponding DNCB control animals. Group mean dermal scores were also noted to be higher In the DNCB test animals as compared to those of the DNCB control animals.

The majority of sensitization study animals gained weight during the test period and generally appeared in good health.

 

In this study, the test item is not a dermal sensitizer.