4-((3-acetoxypropyl)amino)-2,2-dimethyl-4-oxobutane-1,3-diyl diacetate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1998.06.09 tp 1998.07.09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPP 81-6 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

Type of study:

Buehler test

Justification for non-LLNA method:

The study was performed based on the method of BUEHLER EV (1965): Delayed Contact Hypersensitivity in the Guinea Pigs. Arch Dermat 91: 171-177

Test material

Test material information

Constituent 1

Reference substance name:

Panthenol , DL-form

EC Number:

240-540-6

EC Name:

Panthenol , DL-form

Cas Number:

16485-10-2

Molecular formula:

C9H19NO4

IUPAC Name:

2,4-dihydroxy-N-(3-hydroxypropyl)-3,3-dimethylbutanamide

Test material form:

liquid: viscous

Remarks:

At room temperature: high viscous oil, colourless (crystallized)

Details on test material:

Batch No: LJ. No 28740-94 B1 and B2
Date of manufacturing: January 26, 1998

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
female Pirbright White, Dunkin Hartley Crl:(HA)BR [SPF] guinea pigs
- Source: Charles River GmbH, Sulzfeld, FRG
- Age at study initiation: young adult animals, no further data
- Weight at study initiation: 323 - 374 g
- Housing: 5 animals per cage, in stainless steel wire mesh cages with plastic-coated grating, floor area 40 cm x 51 cm
- Diet: Kliba Laboratory Diet (Rabbit- Guinea Pig Maintenance Diet; Supplier: Klingentalmühle AG, Kaiseraugst, Switzerland), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

0.5 mL of the test substance applied to each animal

Day(s)/duration:

days 0, 7 and 14 on teh same application area

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

0.5 mL of the test substance was applied for 6 hours to the right flank of each animal

Day(s)/duration:

14 days after the third induction

No. of animals per dose:

Number of animals per control group : 10
Number of animals of the test group : 20

Details on study design:

INDUCTION
3 inductions were conducted
Application frequency:
• one application per week ; days 0, 7 and 14 on the same application area
2 x 2 cm gauze patches (6 layers surgical gauze Ph . Eur. from Lohmann GmbH & Co.
KG) containing the test substance were applied to the skin of the flank under an
occlusive dressing . The dressing consisted of rubberized linen patches (4 x 4 cm from
Russka), patches of Idealbinde (5 x 5 cm from Pfälzische Verbandstoff-Fabrik) and
Fixomull® Stretch (adhesive fleece) from Beiersdorf AG.
0.5 ml of the test substance was applied to each animal .
The control groups were not treated, since the test substance was applied unchanged
and thus no vehicle was used .
Duration of exposure :
• 6 hours
Site of application:
• anterior left flan k
Reading:
• 24 h after the removal of the patc h

CHALLENGE
A challenge was carried out 14 days after the third induction .
2 x 2 cm gauze patches (6 layers surgical gauze Ph . Eur. from Lohmann GmbH & Co.
KG) containing the test substance were applied to the skin of the flank under an
occlusive dressing . The dressing consisted of rubberized linen patches (4 x 4 cm from
Russka), patches of Idealbinde (5 x 5 cm from Pfälzische Verbandstoff-Fabrik) and
Fixomull® Stretch (adhesive fleece) from Beiersdorf AG .
0.5 ml of the test substance was applied for 6 hours to the right flank of each animal .
The test group and control group 1 were treated with the test substance (control group 2
remained untreated).
Readings :
• 24 and 48 h after the removal of the patch

Positive control substance(s):

no

Remarks:

A positive control (with Alpha-Hexylcinnamaldehyde techn. 85%) is not included in this study but a separate study is performed twice a year in the laboratory.

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Undiluted

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Undiluted

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

Undiluted

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

Undiluted

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Results 5

Reading:

other:

Group:

positive control

Dose level:

Not specified

Clinical observations:

Not specified

Remarks on result:

positive indication of skin sensitisation

Any other information on results incl. tables

Main study:

Induction:

The first, second and third induction with the undiluted test substance did not cause any signs of skin irritation in all test group animals.

Challenge:

The number of animals with skin findings after the challenge is summarized in the following table:

Group	Challenge (Test substance undiluted) skin reactions
Control group 1	0/10
Test group	0/20

x/y: number of positive reactions/number of animals tested (reading at 24 h and/or 48 h after the removal of the patch)

Body weights:

The expected body weight gain was generally observed in the course of the study.

Conclusion:

Based on the results of this study and applying the evaluation criteria it was concluded that D,L-Panthenol does not have a sensitizing effect on the skin of the guinea pig in the BUEHLER Test under the test conditions chosen.

Positive control study (BASF report No. 32H0158/982065)

The positive control with Alpha-Hexylcinnamaldehyde techn. 85% showed that the chosen guinea pig strain was able to detect sensitizing compounds under the laboratory conditions chosen.

The number of animals with skin findings after the challenge and after the rechallenge is summarized in the following table:

Challenge

Rechallenge

a-Hexylcinnamaldehyde 10% in Lutrol E 400 DAB

Vehicle control Lutrol E 400 DAB

a-Hexylcinnamaldehyde 10% in Lutrol E 400 DAB

Vehicle control Lutrol E 400 DAB

24 h

48 h

Total

24 h

48 h

total

24 h

48 h

Total

24 h

48 h

total

Control group 1

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

Control group 2

No application of  test substance

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

0/10

Test group

4/19

2/19

4/19

0/19

0/19

0/19

5/19

3/19

5/19

0/19

0/19

0/19

x/y: number of positive reactions/number of animals tested (reading at 24 h and/or 48 h after the removal of the patch)

(*) 1 animal of the test group died 25 days after the beginning of the study. Macroscopic examination revealed that the cause of death was not substance-related.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the evaluation criteria the results of this study show that DL-Panthenol does not have a sensitizing effect on the skin of the guinea pig in the BUEHLER Test under the test conditions chosen.

Executive summary:

DL-Panthenol was tested for its sensitizing effect on the skin of the guinea pig in the BUEHLER Test according to OECD 406 and under GLP.

The first, second and third induction with the undiluted test substance did not cause any signs of skin irritation in all test group animals.

A challenge was performed 14 days after the last induction.

The number of animals with skin findings after the challenge is summarized in the following table:

Group	Challenge (Test substance undiluted) skin reactions
Control group 1	0/10
Test group	0/20

x/y: number of positive reactions/number of animals tested (reading at 24 h and/or 48 h after the removal of the patch)

Based on the results of this study and applying the evaluation criteria it was concluded that DL-Panthenol does not have a sensitizing effect on the skin of the guinea pig in the BUEHLER Test under the test conditions chosen.