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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental reports

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report Date:
1981

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
The Kligman guinea pig maximization test was conducted using 10 albino Dunkin/Hartley guinea pigs for test chemical Phenethyl salicylate (CAS No: 87-22-9).
GLP compliance:
not specified
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- IUPAC name: 2-phenylethyl 2-hydroxybenzoate
- Common name: Phenethyl salicylate
- Molecular formula: C15H14O3
- Molecular weight : 242.2726 g/mol
- Smiles notation : c1ccc(cc1)CCOC(=O)c2ccccc2O
- InChl: 1S/C15H14O3/c16-14-9-5-4-8-13(14)15(17)18-11-10-12-6-2-1-3-7-12/h1-9,16H,10-11H2
- Substance type: Organic
- Physical state: Solid
Specific details on test material used for the study:
- IUPAC name: 2-phenylethyl 2-hydroxybenzoate
- Common name: Phenethyl Salicylate
- Molecular formula: C15H14O3
- Molecular weight : 242.2726 g/mol
- Smiles notation : c1ccc(cc1)CCOC(=O)c2ccccc2O
- InChl: 1S/C15H14O3/c16-14-9-5-4-8-13(14)15(17)18-11-10-12-6-2-1-3-7-12/h1-9,16H,10-11H2
- Substance type:Organic
- Physical state: Solid
- Physical apperance: white crystals

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Albino Dunkin/Hartley
Sex:
male/female
Details on test animals and environmental conditions:
- Weight at study initiation: 320 g
- Diet (e.g. ad libitum): RGP pellets, hay cabbage ad libitum
- Water (e.g. ad libitum): water ad libitum

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: 6% acetone/20% PEG400/0.01% Tween 80/saline
Concentration / amount:
•two 0.1 ml injections of 0.5% phenethyl salicylate in 6% acetone/20% PEG400/0.01% Tween 80/saline
•two 0.1 ml injections of 0.5% phenethyl salicylate in 50% FCA; and
•two 0.1 ml injections of 50% FCA
Day(s)/duration:
48 hours
Adequacy of induction:
not specified
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
50% phenethyl salicylate in acetone
Day(s)/duration:
24 hours
Adequacy of induction:
not specified
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
10% phenethyl salicylate in acetone
Day(s)/duration:
24 hours
Adequacy of challenge:
not specified
No. of animals per dose:
10 (6 males and 4 females or vise versa)
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE: INTRADERMAL APPLICATION
- No. of exposures: 1
- Exposure period: 7 days
- Test groups: 10
- Control group: no data
- Site: 2X4 cm area of skin in the dorsal shoulder region
- Frequency of applications: no data
- Duration: 1 week
- Concentrations:
• two 0.1 ml injections of 0.5% phenethyl salicylate in 6% acetone/20% PEG400/0.01% Tween 80/saline
• two 0.1 ml injections of 0.5% phenethyl salicylate in 50% FCA; and
• two 0.1 ml injections of 50% FCA

A. II.INDUCTION EXPOSURE: TOPICAL APPLICATION
- No. of exposures: 1
- Exposure period: 48 hours
- Test groups: 10 guinea pigs
- Control group: no data
- Site: same shoulder injection site
- Frequency of applications: no data
- Duration: 48 hours
- Concentrations: 50% phenethyl salicylate in acetone



B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: after 13-14 days
- Exposure period: 24 hours
- Test groups: 10 guine pigs
- Control group: 4 guinea pigs
- Site: clipped and shaved flank
- Concentrations: 10% phenethyl salicylate in acetone
- Evaluation (hr after challenge): 24 and 48 hours

Second challenge was conducted 1 week after on the opposite flank exactly as per the primary challenge. Further challenges and cross challenges maybe made on alternate flanks at weekly or greater intervals as required
Challenge controls:
Two types of controls are used
Treated control: they are given a mock induction treatment at the same time and in the same way as for the test animals except that test substance is omitted from the injection and application preparations. (4 same sex)

Untreated control: at every challenge in the test 4 previously –untreated animals of the same sex and weighing approximately the same as the test animals at the challenge are treated inexactly the same as the test animals.
Positive control substance(s):
not specified

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
other: Challenge 1
Hours after challenge:
24
Group:
test group
Dose level:
10%
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
Weak sensitizer
Remarks on result:
positive indication of skin sensitisation
Reading:
other: Challenge 1
Hours after challenge:
48
Group:
test group
Dose level:
10%
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
Weak sensitizer
Remarks on result:
no indication of skin sensitisation
Reading:
other: Challenge 2
Hours after challenge:
24
Group:
test group
Dose level:
10%
No. with + reactions:
4
Total no. in group:
10
Clinical observations:
Weak sensitizer
Remarks on result:
positive indication of skin sensitisation
Reading:
other: Challenge 2
Hours after challenge:
48
Group:
test group
Dose level:
10%
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
Weak sensitizer
Remarks on result:
positive indication of skin sensitisation

In vivo (LLNA)

Cellular proliferation data / Observations:
Sensitization reactions were observed after both challenges.

Any other information on results incl. tables

Table 1.Skin Reaction Scoring System

SKIN REACTIONS

SCORES

No reaction

0

Barely perceptible erythema

±

Scattered, mild erythema (faint pink)

+

Moderate and diffuse erythema (pale pink )

++

Intense erythema (deep pink) and oedema

+++

 

 

 

Table 2.1.RESULTS OF CHALLENGE 1 (CONCENTRATION: 10%)

Scores

(Hours)

No. of animals

1

2

3

4

5

6

7

8

9

10

Sex

F

F

F

F

F

F

M

M

M

M

24 HOURS

±

±

++

±

+++

±

+

±/+

±

+

48 HOURS

±

±

++

0

+++

0

+

++

±

±

No of positive reactions

24 hours:3+?2

48 hours:2+?1

 

Table 2.2 Treated Controls of Challenge 1(Concentration: 10%)

 Scores

(Hours)

No. of animals

1

2

3

4

Sex

F

F

F

F

24 HOURS

0

0

±

0

48 HOURS

0

0

0

0

 

 

Table 2.3. Untreated Controls of Challenge 1 (Concentration: 10%)

 

 Scores

(Hours)

No. of animals

1

2

3

4

Sex

F

F

F

F

24 HOURS

0

0

0

±

48 HOURS

0

0

±

±

 

 

Table 3.1.RESULTS OF CHALLENGE 2 (CONCENTRATION: 10%)

Scores

(Hours)

No. of animals

1

2

3

4

5

6

7

8

9

10

Sex

F

F

F

F

F

F

M

M

M

M

24 HOURS

+/++

±

++

±

++/+++

0

++

+

0

+

48 HOURS

±/+

0

++

0

+++

0

+

±

0

±

 

Table 3.2. Untreated Controls of Challenge 2 (Concentration: 10%)

 

 Scores

(Hours)

No. of animals

1

2

3

4

Sex

F

F

F

F

24 HOURS

0

0

0

±

48 HOURS

0

0

±

±

 

 

Applicant's summary and conclusion

Interpretation of results:
other: sensitizing
Conclusions:
The test material Phenethyl salicylate (CAS No: 87-22-9) was considered to be sensitizing on albino Dunkin/Hartley guinea pigs in the Kligman guinea pig maximization test.
Executive summary:

The Kligman guinea pig maximization test was conducted using 10 albino Dunkin/Hartley guinea pigs for test chemicalPhenethyl salicylate(CAS No:87-22-9).

 

Preliminary irritation tests are carried out in guinea pigs to determine the concentration of test substances suitable for induction of sensitization and for sensitization challenge.As a result of the preliminary studies, the concentration selected for skin sensitization test were 0.5% forintradermal induction, 50% for topical induction and 10% for challenge.

 

Induction consisted of intradermal injections followed 1week later by a 48 h occluded patch. Three pairs of intradermal injections were made as follows: two 0.1 ml injections of 0.5% phenethyl salicylate in 6% acetone/20% PEG400/0.01% Tween 80/saline; two 0.1 ml injections of 0.5% phenethyl salicylate in 50% FCA; and two 0.1 ml injections of 50% FCA. Seven days later, an occluded patch with 50% phenethyl salicylate in acetone was applied over the shoulder injection sites for 48 h. After a 14-day rest period, the animals were challenged with 10% phenethyl salicylate in acetone applied for 24 h under occlusion using an 11 mm aluminum patch test cup. Reactions were read 24 and 48 h after patch removal.

 

Second challenge was conducted 1 week after on the opposite flank exactly as per the primary challenge. Further challenges and cross challenges maybe made on alternate flanks at weekly or greater intervals as required with controls included.

 

Sensitization reactions were observed after both challenges. Cross-challenge applications with 10% benzyl salicylate and 10% phenyl salicylate were then made at weekly intervals. Cross-reactions were observed after each challenge.

 

Approximately 3 out of 10 guinea pigs showed positive skin sensitization reaction after 2 challenges. Hence the chemicalPhenethyl salicylate(CAS No:87-22-9) was considered to as weak sensitizer on albino Dunkin/Hartley guinea pigs.