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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:
May - Oct 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
This study on skin sensitization was conducted in 1991, prior to current REACH requirements and testing strategies. It is considered adequate for evaluation of this toxicological endpoint.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Test material form:
solid
Specific details on test material used for the study:
Information from study report:
- Name of test material: Emulgator E30, Fest
- CAS number: 5896-54-8
The CAS number in report (5896-54-8) is misleading, since this CAS number does only represent 1-Penta-decanesulfonic acid, sodium salt (1:1), but it is confirmed that the substance used is correctly assigned to CAS 68188-18-1.
- Appearance/Further information: The test substance was supplied as white wax-like leaf-let and was received at the test institute on March 06, 1991.
- Batch No. of test material: 210291
- Purity: 95 %
- Analytical reference: TGL 39237
- Solubility and stability of the test substance in the solvent/vehicle Petrolateum: No data

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Dunkin Hartley Pirbright White; Bor: DHPW (SPF)
- Source: F. Winkelmann, Versuchstierzucht GmbH & Co KG, Borchen, Germany
- Weight at study initiation (mean): 332.2 g ± 19.28
- Housing: 2-4 per cage, Makrolon type IV cages
- Diet and water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): approx. 35-55
- Air changes (per hr): 12-15
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
water
Concentration / amount:
1 % / 0.1 mL
Day(s)/duration:
Day 0
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
1 % / 0.5 g
Day(s)/duration:
Day 7 / 48 hours
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
0.1 % / 0.2 g
Day(s)/duration:
Day 21 / 24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
22
Details on study design:
RANGE FINDING TESTS:
Preliminary range finding study: Three animals, each receiving intradermal injections at concentrations of 0.2 %, 1.0 %, 5.0 % and 10.0 % w/w of the test substance dissolved in water for injection.
Second range finding study (performed twice): 1. Ten animals, each receiving topical applications at concentrations of 0.5 %, 1.0 %, 4.0 % w/w of the test substance in vaseline (= petrolatum). Due to the irritative effects at 24 and 48 hours after patch removal, a second range finding study was carried out. 2. Ten animals, each receiving topical applications at concentrations of 0.1, 0.01 and 0.05 % w/w in vaseline.

MAIN STUDY
On the day before study start, the backs of the animals were carefully shaved to prevent mechanical irritation of the skin. 22 animals were used each for test and control group. An initial intradermal exposure was employed (Day 0), followed by a topical exposure after 7 days (Day 7). Fourteen days later (Day 21), animals were once challenged by topical exposure.

A. INDUCTION EXPOSURE
Intradermal induction (Day 0)
- No. of exposures: Three pairs of intradermal injections were made once into the shoulder region.
a) FCA (Freud's Complete Adjuvant) 0.1 mL, 1:1 diluted in water for injection
b) Test substance formulation 0.1 mL
c) Test substance formulation including FCA 0.1 mL
- Control group: The control group received the vehicle instead of the test substance formulation.
- Site: Injections were made into the right and left shoulder.
- Concentrations: 0.1 mL of the test substance was injected as a 1 % aqueous suspension.
- Evaluation: The test sites were inspected after 1 and 24 hours.

Topical induction (Day 7)
- No. of exposures: once
- Exposure period: 48 hours (occlusive conditions; the patch was tightly wrapped using adhesive tape (Blenderm), Lastohaft and Leukoplast)
- Control group: The control group received the vehicle instead of the test substance formulation.
- Site: Topical applications were made on right and left shoulder.
- Concentration: 0.5 g of the 1 % test substance in vaseline were placed onto 2 x 4 cm filter papers and applied to the shoulder skin.
- Evaluation: The test sites were inspected 1 and 24 hours after removal of the patches.

B. CHALLENGE EXPOSURE
- No. of exposures: once
- Exposure period: 24 hours (occlusive conditions)
- Control group/Test group: The animals of test and control group received the vehicle on the right flank and the test substance formulation on the left flank.
- Site: Topical applications were made on right and left shoulder.
- Concentrations: 0.2 g of the 0.1 % test substance in vaseline were applied to 2 x 2 cm pieces of filter paper in the same fashion as was used for topical induction and put on the left flank.
- Evaluation (hr after challenge): The test sites were inspected 24 and 48 hours after removal of the patches.
Challenge controls:
22 animals were used for challenge control. Each of them received three intradermally injections of
a) FCA (Freud's Complete Adjuvant) 0.1 mL, 1:1 diluted in water for injection, b) Vehicle and c) Vehicle including FCA 0.1 mL, followed by one topical application on Day 7 with 0.5 g of vaseline (= petrolateum) and a topical application for Challenge on Day 21 with 0.1 % test substance in vaseline. Evaluation was 24 and 48 hours after removal of the patches.
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:
48
Group:
test group
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
22
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
22
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
22
Clinical observations:
none
Remarks on result:
other: no indication of skin irritation
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
22
Clinical observations:
none
Remarks on result:
other: no indication of skin irritation

Any other information on results incl. tables

Starting on day 2 after intradermal injection, the control animals exhibiteded red and swollen injection areas. Between day 2 and day 9 these areas opened; this was considered to be a reaction to the intradermal injection of the FCA. Later these areas scabbed and healed. On day 1, all animals receiving the substance showed wounds, swollen, red or partial necrotic injection areas. The stronger reactions, compared to the control animals, was attributed to the test substance's primary irritation. On day 9, after removal of the patch for second induction, the treated' skin on the shoulder area reacted the same in both groups. Therfore it was concluded that the test substance can cause primary dermal irritations, even at concentrations as low as 1%. When challenged with 0.1% test substance no animal showed any irritation reaction or delayed sensitization reaction.

Applicant's summary and conclusion

Executive summary:

For the assessment of skin sensitization a Guinea Pig Maximation test according to OECD TG 406 is available for the substance. In this study 22 animals were used each for test and for control group. Based on a prior range-finding test the concentrations for the main study were set at 1 % for intradermal and topical induction and at 0.1 % for challenge exposure. The vehicles used were sterile water for the intradermal injections and petrolateum for the topical applications.

After challenge, the test substance was well tolerated by the guinea pigs. Neither irritation reactions nor a potential for delayed skin sensitisation reactions was seen, thus the substance was concluded as having no skin sensitising potential based on this assay.