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Administrative data

Description of key information

The test substance was not sensitizing in a Buehler test and a Guinea pig maximization Test (GPMT).

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:
weight of evidence
Study period:
14 Nov. - 22 Nov. 1983
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Remarks:
GLP compliant study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
17 July 1992
Deviations:
yes
Remarks:
no positive control, epicutaneous induction with a non-irritatnt concentration
Qualifier:
according to guideline
Guideline:
other: EPA Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Humans and domestic animals § 81-6 "Dermal sensitization study"
Version / remarks:
November 1992
GLP compliance:
yes
Remarks:
Quality Assurance statement available, 30 Jan. 1984
Type of study:
Buehler test
Justification for non-LLNA method:
LLNA method was not available yet by the time the study was conducted
Species:
guinea pig
Strain:
other: Pirbright-White guinea pigs
Remarks:
Hoe: DHPK (SPFLac)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG
- Females (if applicable) nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: SPF breeding colony
- Age at study initiation: not specified
- Weight at study initiation: average weight 263.2 g
- Housing: group housing
- Diet (e.g. ad libitum): ERKA 9300 complete guinea pig diet, ad libitum
- Water (e.g. ad libitum): tap water in platic bottles, ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 55 +/- 10
- Photoperiod (hrs dark / hrs light): 12/12
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Remarks:
0.9%
Concentration / amount:
50% / 0.5 ml
Day(s)/duration:
Days 1, 3, 5, 8, 10, 12, 15, 17, 19 / 6h each
Adequacy of induction:
other: highest non-irritant concentration
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Remarks:
0.9%
Concentration / amount:
50% / 0.5 ml
Day(s)/duration:
Day 37 / 6h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 animals in treatment group
10 animals in control group
Details on study design:
RANGE FINDING TESTS: Dermal-occlusive application of 3 concentrations (0.5%, 5%, 50% in 0.9% physiological saline) to the flank of 2 animals; treatment for 24h

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 9
- Exposure period: 6h
- Control group: 0.5 ml of physiological saline alone
- Site: Left flank
- Frequency of applications: 2d interval between treatment
- Duration: until day 35
- Concentrations: 50%

B. CHALLENGE EXPOSURE
- No. of exposures: single treatment
- Day(s) of challenge: Day 37
- Exposure period: 6h
- Test groups: 0.5 ml of a 50% solution
- Control group: 0.5 ml of 0.9 ml physiological saline
- Site: Right flank
- Evaluation (hr after challenge): 24h, 48h

Positive control substance(s):
no
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Key result
Group:
positive control
Remarks on result:
other: no positive control
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
23 Oct. - 14 Nov. 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP compliant study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
17 July 1992
Deviations:
yes
Remarks:
epicutaneous induction performed with a non-irritant concentration
Qualifier:
according to guideline
Guideline:
other: EPA Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Humans and domestic animals § 81-6 "Dermal sensitization study"
Version / remarks:
November 1982
Qualifier:
according to guideline
Guideline:
other: Agricultural chemicals, Laws and Regulations Japan, MAFF
Version / remarks:
1985
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
31 July 1992
GLP compliance:
yes (incl. QA statement)
Remarks:
Quality Assurance Statement available, 15 Jan. 1996
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
LLNA method was not available yet by the time the study was conducted
Species:
guinea pig
Strain:
other: Pirbright-White
Remarks:
Hoe, DHPK (SPFLac)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG, Kastengrund, SPF breeding colony
- Females (if applicable) nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: SPF
- Weight at study initiation: average weight 358 g
- Housing: in groups of 5
- Diet (e.g. ad libitum): ssniff Ms-H (V2233), ad libitum
- Water (e.g. ad libitum): tap water in plastic bottles, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 50 +/- 20
- Photoperiod (hrs dark / hrs light): 12 hours
Route:
intradermal
Vehicle:
other: isotonic saline or 50% Freund´s adjuvant
Remarks:
in addition the adjuvant was tested solely
Concentration / amount:
1% / 0.1 ml
Day(s)/duration:
Day 1
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: isotonic saline or 50% Freund´s adjuvant
Remarks:
in addition the adjuvant was tested solely
Concentration / amount:
50% / 0.5 ml
Day(s)/duration:
Day 8 / 48 h
Adequacy of induction:
other: highest non-irritant concentration
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
50% / 0.5 ml
Day(s)/duration:
Day 22 / 24h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20
Details on study design:
RANGE FINDING TESTS:
1. Determination of primary non-irritant concentration with two animals: left flank treated with 1%, 5%, 25%, 50% in isotonic saline, 0.5 ml of test substance preparation was applied to a cellulose patch (2 * 2 cm);
24h treatment; Erythema and oedema scoring 4h after patch removal.
2. Determination of the tolerance of intradermal injections with 3 animals : Preparations (0.2%, 1%, 5%) injected twice (0.1 ml) within a dorsal area measuring 2*4 cm in the vicinity of the shoulder at three injection sites. Examination for local tolerance at 24, 48, 72, 96 h.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: two intradermal injections, one dermal induction treatment
- Test groups:
- Control group: intradermal induction with Freunds´s complete adjuvant and isotonic saline; dermal induction with isotonic saline alone
- Site: intradermal injection within a dorsal area in the viscinity of the shoulder, dermal patch applied at the injection site
- Duration: intradermal induction treatment from day 1-7; dermal induction on day 8
- Concentrations: intradermal 1%, dermal 50% in vehicle

B. CHALLENGE EXPOSURE
- No. of exposures: single exposure
- Day(s) of challenge: day 22
- Exposure period: 24 h
- Test groups: 50% of test material preparation in isotonic saline
- Control group: same treatment as test animals
- Site: left flank
- Concentrations: 50% preparation in vehicle
- Evaluation (hr after challenge): 24, 48 h (days 24, 25)
Positive control substance(s):
yes
Remarks:
benzocain (1% in semi-liquid paraffin for intradermal induction and 25% concentration in petrolatum for dermal induction and challenge treatment), Report no. 95.0578
Positive control results:
positive indication of sensitization for the positive control substance
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no erythema or oedema
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25% benzocain
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
instances of moderate to severe erythema, slight oedema
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25% benzocain
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
instances of moderate to severe erythema, very slight oedema
Remarks on result:
positive indication of skin sensitisation

Induction treatment

The intradermal injections with Freund's Adjuvant (with and without test substance) caused severe erythema and oedema as well as indurations and encrustations. The application sites treated with the test substance showed slight erythema. Intradermal injections of the vehicle alone caused no signs of irritation.

After the removal of the patches at day 10, severe erythema and oedema, indurated and encrusted skin were observed at the sites previously treated with Freund's Adjuvant. The application sites treated with the test substance showed slight erythema and oedema. The vehicle alone showed no signs of irritation.

Challenge treatment

The body weight gain of the animals was not impaired throughout the study. There were no clinical signs of intoxication. After challenge treatment no signs of irritation occured.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The sensitizing potential of the test substance has been evaluated in a Buehler test and a GPMT.

In the Buehler test 20 female Pirbright White guinea pigs were treated with the test substance. Additional animals were used as negative controls (10 females). The induction of sensitisation was conducted by close-patch topical applications of test substance. (0.5 ml of a 50% dilution in 0.9% physiological saline, placed under occlusive patches for 6 hrs). The test article was applied 3 times per week with a 2-d interval, for 3 weeks. The animals were challenged with the test article (0.5 ml of a 50% dilution in 0.9% physiological saline, placed under occlusive patches for 6 hrs) 2 weeks after the induction phase by close-patch topical application. The resulting reactions were evaluated at 24 and 48 hrs after the challenge treatment.

No clinical signs of intoxication were observed at any time during the study. Body weight gains showed no discernible differences between control and treatment animals. After challenge treatment no signs of irritation occurred in the treatment and control group.

In the GPMT, 20 female Pirbright-White guinea pigs were treated with test substance. 10 females were used as negative controls and 5 females for determination of the tolerance of intradermal injections. The induction of sensitisation was carried out by two intradermal injections followed by a dermal application one week later. Intradermal injections of the treatment group was performed with the following preparations: 0.1 ml 1% test substance in isotonic saline, 0.1 ml 50% Freund´s Complete Adjuvant in physiological saline, and 0.1 ml 1% test substance in 50% Freund´s Original Adjuvant. Dermal application of the treatment group was performed with 0.5 ml of 50% test substance in isotonic saline, placed under occlusive patches for 48 hrs. The animals were challenged with 0.5 ml of a 50% dilution of test substance in isotonic saline 2 weeks after the induction phase by close-patch topical application (placed under occlusive patches for 24 hrs). The resulting reactions were evaluated at 24 and 48 hrs after the challenge treatment. A positive test substance, benzocain (1% concentration in semi-liquid paraffin for intradermal induction and 25% concentration in petrolatum for dermal induction and challenge treatment) was tested according to the same procedure in a preliminary experiment.

There were no clinical signs of intoxication in the main study. Body weight gains showed no discernible differences between control and treatment animals. Intradermal injections with Freund´s Adjuvant (with and without test substance) caused severe erythema and oedema as well as indurations and encrustations. The application sites treated (intradermal injections) with the test substance showed slight erythema. Intradermal injections of the vehicle alone caused no signs of irritation in control animals. After challenge treatment, no signs of irritation occurred in the treatment and control group. The positive control substance gave the expected positive response.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the data available and in accordance with Regulation (EC) 1272/2008 (CLP), non-classification for skin sensitization is warranted.