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Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 October to 16 November 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study undertaken at GLP accredited laboratory to internationally accepted guidelines.
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Species:
rabbit
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Commercial laboratory animal supplier.
- Age at study initiation: 5 weeks
- Weight at study initiation:
- Housing: in a mobile battery, containing 6 cages; maximal 10 animals per cage.
- Diet (e.g. ad libitum): standard laboratory diet ad libitum.
- Water (e.g. ad libitum): Tap water (N.V. Hydron Midden Nederland) ad libitum.
- Acclimation period:12 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3°C
- Humidity: 30 - 70%
- Air changes: ca. 10 air changes per hour.
- Photoperiod: 12 hrs dark / 12 hrs light

Route:
intradermal
Vehicle:
maize oil
Concentration / amount:
Concentration of test material and vehicle used at induction: Induction treatment by intradermal injections of Freund's Complete Adjuvant (FCA) 1:1 dilution with physiological saline, a 0.3 % test dilution of the test substance in maize oil, and a 0.3 % test dilution of the test substance in FCA/maize oil, followed one week later by topical application of a 30 % test dilution of the test substance in maize oil. Concentration of test material and vehicle used for each challenge: 14 days after the last induction, by topical application of a 3 % dilution of the test substance in maize oil and maize oil alone.
Route:
intradermal and epicutaneous
Vehicle:
maize oil
Concentration / amount:
Concentration of test material and vehicle used at induction: Induction treatment by intradermal injections of Freund's Complete Adjuvant (FCA) 1:1 dilution with physiological saline, a 0.3 % test dilution of the test substance in maize oil, and a 0.3 % test dilution of the test substance in FCA/maize oil, followed one week later by topical application of a 30 % test dilution of the test substance in maize oil. Concentration of test material and vehicle used for each challenge: 14 days after the last induction, by topical application of a 3 % dilution of the test substance in maize oil and maize oil alone.
No. of animals per dose:
Number of animals in test group: 10
Number of animals in negative control group: 5
Details on study design:
RANGE FINDING TESTS:
The irritation response to intradermal injection of various concentrations of the test substance was examined in 2 guinea pigs. A sufficiently large area ofthe flanks was clipped free from hair with electric clippers. Amounts of 0.1 ml of the selected concentrations were applied by intradermal injection. Circa 24 hours after injection, the animals were examined for signs of irritation. A concentration causing slight to moderate irritation but otherwise well-tolerated by the animals, is usually taken for intradermal injection of the test substance in the induction phase of the main study. The irritation response to topical treatment ofvarious concentrations of the test substance was examined in 2 other guinea pigs. The flanks of each of the animals were clipped free from hair with electric clippers. Patches (Silverpatch, v.d. Bend B.V., nBrielle, the Netherlands) were loaded with the test material and placed on the clipped skin of each animal, and covered with a piece of hypoallergenic paper bandage (Leukopor) that was secured by elastic adhesive bandage (Tensoplast), wound around the torso of the animal. The dressing was left in place for ca. 24 hours. Circa twenty four and 48 hours after removal of the dressing, the animals were examined for signs of skin irritation. A concentration causing slight to moderate skin irritation is usually chosen for topical induction and a non-irritant concentration for topical challenge.

B. CHALLENGE EXPOSURE
Fifteen guinea pigs were randomly divided into two groups, viz. one test group of 10 animals and one control group of 5 animals. The animals were weighed one day before the study was initiated and at the completion ofthe study.

Induction was effected in two different ways, firstly by intradermal injections and secondly, one week later, by topical application over the injection sites.

a. Intradermal injections
For this purpose an area of about 24 cm2 of dorsal skin in the scapular region was clipped free from hair with electric clippers. Pairs of intradermal injections (0.1 ml each) were made simultaneously in the clipped area.
The following preparations were injected:
test animals
- two injections with Freund's Complete Adjuvant (FCA)/physiological saline (1: 1),
- two injections with the selected test concentration,
- two injections with the selected test concentration in FCA Ivehicle (1:1),
control animals
- two injections with FCAIphysiological saline (1: 1),
- two injections with the vehicle,
- two injections with FCAIvehicle (l: 1).
Skin readings were made at ca 24 hours after the treatment.

b. Topical application
Six days after the intradermal injections, the dorsal skin in the scapular region of all test and control animals was closely clipped again. On the following day, the induction by topical application was made in this region. The test animals were treated as follows: A circa 2 x 4 cm patch ofWhatman No.3 MMfilter paper was loaded with the selected concentration ofthe test substance (see table 1). The loaded patch was placed over the sites ofthe intradermal injections and was secured as described in section 2.3.1. The dressing was left in place for ca 48 hours. The controls were similarly treated with a patch loaded with the vehicle. Skin readings were made directly after removal ofthe patches.

The topical challenge with the test substance was carried out 14 days (test animals) or 15 days (controls) after the topical induction as follows:
An area ofcirca 5 x 5 cm on both flanks of each test and control animal was clipped free from hair. Patches were loaded with the test concentration selected and with the vehicle alone. Subsequently, the loaded patches were placed on the clipped area of the flank ofeach test and control animal. The patches were covered with Leukopor bandage, and held in place by Tensoplast for ca 24 hours. Skin readings were made at
ca 24 and 48 hours after removal ofthe patches.

Challenge controls:
No data
Positive control substance(s):
no
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 5.0.

Maximum concentration not causing irritating effects in preliminary test: .3 %

Signs of irritation during induction:
The intradermal injections generally caused the following skin reaction:

FCA / physiological saline (1:1): moderate

Vehicle: no skin reactions

FCA / vehicle (1:1): moderate erythema

After the topical application of the vehicle alone, no skin reactions were observed in controls.

Evidence of sensitisation of each challenge concentration:
At 24 and 48 hours after the challenge treatment with the test substance, none of the controls or test animals showed
any skin reaction.

None of the controls or test animals showed any skin reaction at 24 and 48 hours after challenge with the vehicle
alone.

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Since none of the test animals showed positive signs of sensitization, it can be concluded that, according to the EC standards, the test substance is not a sensitizer.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Justification for selection of skin sensitisation endpoint:

Only study available

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The skin sensitising study returned a negative result. Therefore BPS-MAE will not be classified as a sensitiser.