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Description of key information

In a study according OECD TG 406 performed on guinea-pigs the substance is not to be classified as skin sensitizer (reference 7.4.1 -1).

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August 08, 2000 - September 01, 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted on July 1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
1996
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The guinea pig test method described in OECD TG 406 provide suitable information for hazard identification.
Species:
guinea pig
Strain:
other: HsdPoc:DH
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen
- Age at study initiation: about 5 weeks
- Weight at study initiation: mean 349 g (range from 322 to 381 g)
- Housing: Two guinea pigs were housed in a Makrolon cage type with a shelter placed on mobile racks.
- Diet: ad libitum, exclusive diet for guinea pigs No. 845, Eberle Nafag
- Water: ad libitum, tap water
- Acclimation period: at least 7 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 23
- Humidity (%): 48 to 87
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal
Vehicle:
other: Sodium chloride solution (isotonic)
Concentration / amount:
1 g/L (0.1 mL injected)
Day(s)/duration:
7 days
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: Sodium chloride solution (isotonic)
Concentration / amount:
5.0 g/L
Day(s)/duration:
2 days
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: Sodium chloride solution (isotonic)
Concentration / amount:
0.1 g/L
Day(s)/duration:
2 days
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Pretest: 5 females
Group 1: negative control group (10 females)
Group 2: test material group (20 females)
Details on study design:
RANGE FINDING TESTS:
To determine the concentrations suitable for the main study, a pretest with single intradermal or topical administrations of the vehicle and of test material preparations was performed. Intradermal (i.d.) injections were given to one animal. Four further animals were used to determine the topical concentrations for the main study. Two of the guinea pigs were exposed for 48 hours without pretreatment to determine the concentration for the dermal induction. The other two animals were treated according to the control group including intradermal injections with FCA. One week after topical induction the two animals of the pretest group were exposed for 24 hours to different test material concentrations to find out the challenge concentration. In the pretest the following concentrations were used:
1. Sodium chloride solution: intradermal
2. Test item with sodium chloride solution as vehicle:
intradermal: 50, 10, 5, 1, and 0.1 g/L
topical: 200, 100, 60, 30, 15, 10, 7.5 and 1 g/L
topical with FCA: 1.0, 0.5, 0.1, and 0.01 g/L

MAIN STUDY
A. INDUCTION EXPOSURE
Induction stage I (intradermal injection)
- No. of exposures: six intradermal injections were given (three on each side of the spinal column in a total area of 2 x 4 cm).
- Exposure period: 7 days
- Test groups: 1.0 g/L test item in sodium chloride solution + Freund´s complete adjuvant
- Control group: vehicle Sodium chloride solution + Freund´s complete adjuvant
- Site: shoulder region (cranial, medial, caudal)
Group 1 (vehicle: Sodium chloride solution)
cranial: 0.10 mL Freund's complete adjuvant + Sodium cloride solution
medial: 0.10 mL Sodium chloride solution
caudal: 0.10 mL Freund's complete adjuvant + Sodium cloride solution
Group 2 (test material)
cranial: 0.10 mL Freund's complete adjuvant + Sodium cloride solution
medial: 0.10 mL Test material (1.0 g/L sodium chloride solution)
caudal: 0.10 mL Freund's complete adjuvant with test material + Sodium cloride solution (1.0 g test material/L preparation)

Induction stage II (topical application)
One week after the intradermal injections, the shoulder area of the guinea pigs was shaven again and covered with a filter paper patch of about 8 cm2 fully soaked with 1 mL of the test material preparation or the vehicle. The patches were attached for 48 hours under occlusive conditions.
Group 1: Sodium chloride solution
Group 2: Test item (5.0 g/L sodium chloride solution)
During the induction phase, the guinea pigs were examined on days 2, 3, 7, 11, 12, 15, and 22 for local skin reactions, and the results were documented. The behavior and general condition of all animals were monitored daily.

B. CHALLENGE EXPOSURE
- No. of exposures: one topical exposure
- Day(s) of challenge: Two weeks after the topical induction
- Exposure period: 24 hours
- Test groups: 0.1 g/L in sodium chloride solution
- Control group: vehicle Sodium chloride solution
- Site: shaven flanks of the animals
- Evaluation (hr after challenge): The challenge sites were investigated for reactions 48 hours after start of the challenge. Further inspections followed 72 hours after start of the challenge, to detect weak or slowly developing reactions.
Skin changes at the application sites were graded according to the MAGNUSSON and KLIGMAN scheme following the grading scale:
0 = no reaction
1 = scattered mild redness
2 = moderate and diffuse redness
3 = intense redness and swelling



Challenge controls:
yes
Positive control substance(s):
yes
Remarks:
a-Hexylcinnamaldehyde
Positive control results:
Data from the GLP study T14666 (Experimental part: April 11 until May 05, 2000) with 10 g/L alpha-Hexylcinnamaldehyde in Polyethylen glycol 400
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
The clinical behavior of the guinea pigs was normal during the experimental part. All animals survived the experimental part. The body weight development corresponded to that of the animals of the vehicle group.
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
The clinical behavior of the guinea pigs was normal during the experimental part. All animals survived the experimental part. The body weight development corresponded to that of the animals of the vehicle group.
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1 g/L
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The clinical behavior of the guinea pigs was normal during the experimental part. All animals survived the experimental part. The body weight development corresponded to that of the animals of the vehicle group.
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
0.1 g/L
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The clinical behavior of the guinea pigs was normal during the experimental part. All animals survived the experimental part. The body weight development corresponded to that of the animals of the vehicle group.
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
Dose level:
10 g/L
No. with + reactions:
7
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
positive control
Dose level:
10 g/L
No. with + reactions:
4
Total no. in group:
10
Interpretation of results:
GHS criteria not met
Executive summary:

A study was conducted according to OECD TG 406 to evaluate the skin sensitizing properties of the test material in the guinea pig maximization test according to MAGNUSSON and KLIGMAN (1969). 10 female guinea pigs in the negative control group (group 1) treated with the vehicle sodium chloride solution and 20 females in the test material group (group 2) were investigated. Induction included intradermal injection of test material preparation (1.0 g/L with and without Freund's complete adjuvant) on day 1, and topical application of test material preparation (5.0 g/L) on day 8. Challenge by topical application of the test material preparation (0.1 g/L) was performed two weeks after topical induction.

After challenge no positive reactions were observed in the test material treated skin sites.

Endpoint:
skin sensitisation: in vitro
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

A study was conducted according to OECD TG 406 to evaluate the skin sensitizing properties of the test material in the guinea pig maximization test according to MAGNUSSON and KLIGMAN (1969).10 female guinea pigs in the negative control group (group 1) treated with the vehicle sodium chloride solution and 20 females in the test material group (group 2) were investigated.Induction included intradermal injection of test material preparation (1.0 g/L with and without Freund's complete adjuvant) on day 1, and topical application of test material preparation (5.0 g/L) on day 8. Challenge by topical application of the test material preparation (0.1 g/L) was performed two weeks after topical induction. After challenge no positive reactions were observed in the test material treated skin sites. Thus, the test item is not to be classified as skin sensitizer (reference 7.4.1 -1).

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on this information, the substance is not considered to be classified for skin sensitisation under Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EU) 2019/521.