Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
acute toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
According to Regulation (EC) No 1907/2006, Annex VIII, section 8.5, column 2, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route in addition to the oral route (8.5.1) for substances other than gases. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. Testing by the dermal route is appropriate if: (1) inhalation of the substance is unlikely; and (2) skin contact in production and/or use is likely; and (3) the physicochemical and toxicological properties suggest potential for a significant rate of absorption through the skin.

In consideration of the same section (Column 2 of Annex VIII, Section 8.5.3), testing by the dermal route does not need to be conducted if the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route, and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation) or, in the absence of an in vivo study by the oral route, no systemic effects after dermal exposure are predicted on the basis of non-testing approaches (e.g. read-across, QSAR).The target substance Propylene dinonanoate (CAS 41395-83-9) did not cause mortality or adverse clinical signs in an acute toxicity study following oral administration of 5000 mg/kg bw in female mice. In a study conducted in male and female rats an LD50 of 16025 mg/kg was calculated.
Furthermore, no systemic effects were noted in two in vivo skin irritation studies performed with the target substance Propylene dinonanoate (CAS 41395-83-9). No systemic effects were noted in in vivo skin sensitisation studies with the two analogue substances dodecanoic acid, ester with 1,2-propandiol (CAS 37321-62-3) and C8-10 Fatty acids, vegetable-oil, esters with dipropylene glycol (CAS 95009-41-9).
Therefore, testing by the dermal route is not appropriate in accordance with Annex VIII, Section 8.5.3 as the target substance 1) does not meet the classification criteria for acute toxicity based on the data available for the oral route; and 2) no systemic effects of toxicological relevance have been observed in the available in vivo studies with dermal exposure. Hence, testing for acute dermal toxicity should be avoided for reasons of animal welfare especially as no additional knowledge is expected to be gained by an acute dermal toxicity study considering the outcome of the available, relevant studies (including acute oral toxicity and in vivo studies with dermal exposure).
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
No purity of test substance reported.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
not applicable
Principles of method if other than guideline:
Study was conducted prior to adoption of any OECD guideline.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Fasting period before study: yes, 20-24 h
- Housing: 2-5 per cage
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
The test material was administered as supplied by the sponsor.
Doses:
Preliminary test: 1000, 5000, 10000 mg/kg bw
Main test: 10000, 15000, 20000 mg/kg bw
No. of animals per sex per dose:
Preliminary test: 2 at 1000 mg/kg bw, 1 at 5000 and 1 at 10000 mg/kg bw
Main test: 5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
Statistics:
Probit method
Preliminary study:
2m+2f animals at 1000 mg/kg: no mortality
1m+1f animal at 5000 mg/kg: no mortality
1m+1f animal at 10000 mg/kg: no mortality
Key result
Sex:
male/female
Dose descriptor:
LD50
Remarks:
rat
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
male/female
Dose descriptor:
LD50
Remarks:
rat
Effect level:
16 025 mg/kg bw
Based on:
test mat.
Remarks:
Calculated using Probit method
95% CL:
> 12 973 - < 19 795
Remarks on result:
other:
Remarks:
5 out of 10 animals were dead after 14d observation period after single oral dosing at 15000 mg/kg
Mortality:
1 out of 10 rats was found dead after dosing at 10000 mg/kg. No mortality was seen in the preliminary study.

Dose
mg/kg

Gender

Total number of rats

Number of dead rats after 14 day observation period

% mortality

1000

m

2

0

0

1000

f

2

0

0

5000

m

1

0

0

5000

f

1

0

0

10000

m

1

0

0

10000

f

1

0

0

10000

m

5

1

10

10000

f

5

0

0

15000

m

5

4

80

15000

f

5

1

20

20000

m

5

4

80

20000

f

4

2

50

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
13 Mar 1991 - 13 Mar 1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
No purity of test substance reported. Observation period only 7 days after treatment.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
adopted in 1987
Deviations:
yes
Remarks:
No purity of test substance reported. Observation period only 7 days after treatment
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
mouse
Strain:
NMRI
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 19 to 20 g

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5.7 mL/kg
Doses:
5000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 days
- Frequency of weighing: on day of treatment and last day of observation
- Frequency of observations: once daily
Key result
Sex:
female
Dose descriptor:
LD50
Remarks:
mouse
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
No abnormal clinical signs and no abnormal behaviour occured.
Body weight:
All 5 female mice showed a gain in body weight (gain of 7.5 - 8g per mouse over a period of 7 days).
Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
No purity of test substance reported.
Qualifier:
according to guideline
Guideline:
other: Journal Officiel de la Republique Francoise 1971/1973
Deviations:
not specified
Principles of method if other than guideline:
Study was conducted prior to adoption of any OECD guideline.
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 2.5-3.5 kg
- Housing: individual in 600x540x315 mm cages or in restraining devices
- Diet: 200g feed per day, commercial rabbit feed.
- Water: ad libitum
Animals were vaccinated against Myxomatosis and Pasteurellosis. Only healthy animals without signs of skin lesions were included.

ENVIRONMENTAL CONDITIONS
Air conditioned.
Type of coverage:
occlusive
Preparation of test site:
clipped
Remarks:
back of rabbit, 14 cm x 14 cm clipped. Superficial incisions were made to the right flank.
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 mL of undiluted test substance
Duration of treatment / exposure:
23 h
Observation period:
Readings at 24 and 72 hours after start of exposure.
Number of animals:
6
Details on study design:
TEST SITE
- Area of exposure: Test substance was placed on two squares of sterile absorbent gauze measuring 2 cm x 2 cm; each square was composed of four layers of gauze. The gauze was placed on the left (intact skin) and right flank (incised skin) and held in contact with skin with an occlusive patch, which was adhesive and non-allergic. Additional an adhesive tape was wrapped around the animals.

SCORING SYSTEM:
Erythema: 0 (no), 1 (slight), 2 (well defined), 3 (moderate to severe), 4 (severe with slight eschar formation - injury in depth)
Edema: 0 (no), 1 (very slight), 2 (slight - definite raising), 3 (moderate), 4 (severe)
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
animal: #1, #2, #3, #4, #5, #6
Remarks:
individual mean over
Time point:
other: 24h/72h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other:
Remarks:
left flank
Irritation parameter:
erythema score
Remarks:
intact skin
Basis:
animal: #1, #2, #3, #4, #5, #6
Remarks:
individual mean over
Time point:
other: 24h/72h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other:
Remarks:
left flank
Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 to 11 Mar 1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Purity of test substance not reported. No GLP statement. Limited reporting details.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted in 2015
Deviations:
yes
Remarks:
no rationale for in vivo testing, no initial testing with one animal
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted in 1981
Deviations:
yes
Remarks:
Purity of test substance not reported. No GLP statement. Limited reporting details.
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Legard
- Weight at study initiation: 1.97 kg

IN-LIFE DATES: 01 Mar 1991 to 08 Mar 1991
Type of coverage:
occlusive
Preparation of test site:
not specified
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 mL
Duration of treatment / exposure:
4 h
Observation period:
72 h
Number of animals:
3 males
Details on study design:
TEST SITE
- Area of exposure: not stated

REMOVAL OF TEST SUBSTANCE
- Washing: no washing was performed

OBSERVATION TIME POINTS
1, 24, 48, and 72 h

SCORING SYSTEM:
No details reported.
Irritation parameter:
erythema score
Basis:
animal: #1, #2, #3
Remarks:
individual mean over
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility not applicable
Irritation parameter:
edema score
Basis:
animal: #1, #2, #3
Remarks:
individual mean over
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility not applicable
Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02 Apr - 31 Jul 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The used methodology was validated in the test facility according to the performance standards defined in OECD GL 429. A publication was included as reference for the validity of the methodology as required according to OECD 429. The threshold EC1.4 used in the study is defined more stringent compared to the threshold provided in the publication.
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
adopted in 2010
Deviations:
yes
Remarks:
Lymphocyte proliferation was measured based on cell counting instead of dpm hence, a different cut off value (EC1.4 instead of EC3) was used for evaluation of results.
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
2012
GLP compliance:
yes (incl. QA statement)
Remarks:
(GROUPE INTERMINISTERIEL DES PRODUITS CHIMIQUES)
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
other: CBA/J (CBA/J@Rj)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elevage Janvier Labs, Le Genest Saint Isle, France
- Age at study initiation: 8 weeks
- Weight at study initiation: 19.1 - 21.8 g
- Housing: individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes
- Diet: A04, SAFE (Augy, France), ad libitum
- Water: tap water from public distribution system, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
25, 50 and 100%
No. of animals per dose:
4 (+1 additional animal/group as reserve)
Details on study design:
RANGE FINDING TESTS:
In a preliminary screening test, the dorsal surface of each ear from 1 mouse was treated daily with 25 µL undiluted test substance for 3 consecutive days. The mouse was observed for signs of toxicity or excessive local irritation. Ear thickness was recorded on Day 1, Day 3 and Day 6. Bodyweight was recorded on Day 1 (prior to dosing) and Day 6. Application of undiluted test substance did not induce clinical signs of systemic toxicity or cutaneous reactions including ear swelling or induction of erythema. Moreover, no irritation was observed due to test substance application. Therefore, no signs indicative for skin irritation were determined and hence, the undiluted test substance was included in the main study as highest concentration.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: lymphocyte cell counting
- Criteria used to consider a positive response: The proliferative response of lymph node cells was expressed as the number of lymphocytes per lymph node and as ratio of lymphocytes of test nodes relative to that recorded for control nodes :

SI = cell count of treated group/cell count of control group

The test item will be regarded as a sensitiser if at least one concentration of the test item results is
greater than 1.4 compared to control values (SI ≥ 1.4) . Other relevant criteria such as dose-response and irritation level were also taken into account for the interpretation of the results. Any test item failing to produce a SI < 1.4 will be classified as a "non-sensitiser".

TREATMENT PREPARATION AND ADMINISTRATION:
25 µL of the test item was applied on the entire dorsal surface of each ear. The application was repeated once daily over 3 consecutive days. Body weights were recored on Day 1 (prior to dosing) and Day 6 (prior to termination). Local irritation reactions were assessed and ear thickness measurements were performed with a micrometer on Day 1 and Day 3 before application and on Day 6 after sacrifice. Furthermore, punch biopsies of both ears were prepared for weighing on Day 6 after sacrifice. The draining auricular lymph nodes of each ear were excised, weighed and pooled for each experimental group. A single cell suspension was prepared by gentle mechanical tissue disaggregation through a 200-meshcell strainers. 10 µL of this cell suspension was diluted in 10 mL of physiological saline solution before lymphocyte cells were counted using a cell counter (Beckman Coulter Z2). A size range of 5 - 15 µm was selected for counting which covers the average size of a lymphocyte of 8 µm.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Key result
Parameter:
SI
Value:
0.98
Test group / Remarks:
25% test substance
Key result
Parameter:
SI
Value:
1.02
Test group / Remarks:
50% test substance
Key result
Parameter:
SI
Value:
1.32
Test group / Remarks:
100% test substance
Key result
Parameter:
SI
Value:
1.13
Test group / Remarks:
5% positive control substance
Key result
Parameter:
SI
Value:
1.48
Test group / Remarks:
10% positive control substance
Key result
Parameter:
SI
Value:
2.19
Test group / Remarks:
25% positive control substance
Key result
Parameter:
other: EC1.4
Test group / Remarks:
test substance groups
Remarks on result:
not determinable
Remarks:
as all SI values were < 1.4
Key result
Parameter:
other: EC1.4
Remarks:
[%]
Value:
8.86
Test group / Remarks:
positive control substance

No significant lymphoproliferative response was noted for the tested concentrations of Fatty acids, vegetable-oil, esters with dipropylene glycol. The calculated stimulation index values were 0.98, 1.02 and 1.32 at test item concentrations of 25%, 50% and 100%, respectively. Thus, as all SI indexes were below 1.4, an EC1.4 value could not be determined.

The positive control substance induced a significant lymphoproliferative response.

Mortality and clinical signs of toxicity

Application of the test substance did not induce mortaility or clinical signs of systemic toxicity. Body weights of control and test animals were comparable over the study period.

Local skin irritation

No cutaneous reactions were noted during the main test. Dose-dependent increases in ear and lymph node weights were determined which did not result in statistically significant differences. Values obtained for ear thickness were comparable among the groups. Therefore, the test item was not considered as excessively irritant at these concentrations.

Table 2: Cell count. Stimulation index and calculation of EC1.4

 

Group

Cell count/group (x 106cells /mL)

SI

Control (AOO)

36.02

 

25%

35.34

0.98

50%

36.66

1.02

100%

47.38

1.32

 

Table 3: Individual ear thickness

 

Group

Ear thickness (mm)

Increase in ear thickness (%)

Increase in ear thickness (%)

Day 1

Day 3

Day 6

D3/D1

D6/D1

Control (AOO)

0.21

0.22

0.18

4.8

-14.3

0.21

0.22

0.20

4.8

-4.8

0.23

0.23

0.20

0.0

-13.0

0.22

0.23

0.20

3.5

-10.3

Mean

0.22 ± 0.01

0.23 ± 0.01

0.20 ± 0.01

3.5 ± 2.3

-10.3 ± 4.3

25 %

0.20

0.21

0.21

5.0

5.0

0.23

0.23

0.19

0.0

-17.4

0.23

0.23

0.19

0.0

-17.4

0.23

0.23

0.20

0.0

-13.0

Mean

0.22 ± 0.02

0.23 ± 0.01

0.20 ± 0.01

1.3 ± 2.5

-10.7 ± 10.7

50%

0.21

0.21

0.19

0.0

-9.5

0.24

0.22

0.20

-8.3

-16.7

0.22

0.22

0.21

0.0

-4.5

0.24

0.24

0.21

0.0

-12.5

Mean

0.23 ± 0.02

0.22 ± 0.01

0.20 ± 0.01

-2.1 ± 4.2

-10.8 ± 5.1

100%

0.24

0.22

0.21

-8.3

-12.5

0.22

0.22

0.21

0.0

-4.5

0.23

0.23

0.20

0.0

-13.0

0.21

0.21

0.19

0.0

-9.5

Mean

0.23 ± 0.01

0.22 ± 0.01

0.20 ± 0.01

-2.1 ± 4.2

-9.9 ± 3.9

 

Table 4: Individual ear biopsy and lymph node weight

 

Group

Ear weight (Day 6 (mg)

% of control

Lmph nodes (mg)

% of control

Control (AOO)

27.8

 

5.2

 

27.9

4.8

25.8

4.9

25.9

4.5

Mean

26.9 ± 1.2

 

4.9 ± 0.3

 

25 %

26.0

 

5.4

 

27.3

6.5

29.4

4.8

26.8

4.9

Mean

27.4 ± 1.5

2.0

5.7 ± 0.7

16.3

50%

26.4

 

6.4

 

30.0

6.3

27.9

5.9

31.3

5.9

Mean

28.9 ± 2.2

7.6

6.1 ± 0.3

24.5

100%

29.2

 

6.8

 

32.9

6.2

30.0

6.6

28.1

5.7

Mean

28.1 ± 2.1

11.9

6.3 ± 0.5

28.6

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
20 Jun - 02 Sep 1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
No details on reliability check
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted in 1992
Deviations:
yes
Remarks:
no details on reliability check
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was conducted prior to adoption of LLNA method.
Species:
guinea pig
Strain:
Dunkin-Hartley
Remarks:
Pirbright White Bor: DHPW [SPF]
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: young adults
- Weight at study initiation: 390 - 532 g (males)
- Housing: animals were housed in groups of at the most 5 animals in Makrolon IV cages.
- Diet: Ssniff G4 - Alleindiät für Meerschweinchen (Ssniff Spezialfutter GmbH, Soest, Germany), ad libitum
- Water: tap water (Gelsenwasser, Haltern, Germany), ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal
Vehicle:
corn oil
Concentration / amount:
10%
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:
unchanged (no vehicle)
Concentration / amount:
100%
Day(s)/duration:
On day 7 for 48 hours
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100%
Day(s)/duration:
On day 21 for 24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Range finding study:
2 (intradermal application)
4 (epicutaneous application)

Main study:
10 (controls), 20 (in test group)
Details on study design:
RANGE FINDING TESTS:
To assess the intracutaneous tolerability of the test substance 2 animals were treated on the left shoulder region. The intracutaneous treatment was applied to groups of 2 animals in concentrations of 0.25, 0.5, 1.0, 2.5, 5.0 and 10.0% in the vehicle corn oil, and evaluated 24 h post appication. Occlusive dermal application of the test substance was conducted in 4 animals for 24 h in concentrations of 2.5, 25, 50 and 100% of the test substance. Dermal reactions were evaluated 48 and 72 h after application. Results of the prelimimary test are presented in Table 1 under "Any other information on results incl. tables".
Based on the results of the range finding test, concentrations of 10% and 100% were selected for intradermal and epicutanous treatment, respectively.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/NaCl solution
Injection 2: test substance in corn oil
Injection 3: test substance in a 1:1 mixture (v/v) FCA/NaCl solution
Epicutaneous: test substance
- Control group I+II:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/NaCl solution
Injection 2: corn oil
- Site: shoulder region (intradermal + epicutaneous)
- Frequency of applications: every 7 days
- Duration: Days 0-7
- Concentrations: intradermal 10%, epicutaneous 100%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 21
- Exposure period: 24 h
- Test groups: test substance
- Control group I: test substance
- Control group II: vehicle control is employed only in the induction treatment; no challenge with vehicle
- Site: left flank
- Concentrations: 100%
- Evaluation (hr after challenge): 24, 48 and 72 h
SCORING SYSTEM:
Skin reactions (erythema, eschar formation and edema) were evaluated according to the scheme presented in Table 1 under "Any other information on materials and methods incl. tables".

OTHER:
The highest concentration of test substance administered dermally in the preliminary test caused no irritation of the skin. In order to cause slight to moderate inflammation of the skin for the dermal induction, one day before the dermal administration (Day 6) all the test and control animals were sheared on the shoulder and treated with sodium dodecyl sulphate (10%).
Challenge controls:
The control group is actually a challenge control.
Positive control substance(s):
not specified
Remarks:
According to the author, the reliability of the test system is checked at regular intervals (no details provided).
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
induction: 0%; challenge: 100%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
induction: 10% and 100%; challenge: 100%
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
induction: 0%; challenge: 100%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
induction: 10% and 100%; challenge: 100%
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation
Reading:
other: reliability check
Group:
positive control
Remarks on result:
other: According to the author, the reliability of the test system was checked at regular intervals (no details provided).

Table 1. Results of the intradermal application (preliminary test).

Concentration

(w/w)

animal 1

   E   O

animal 2

 E O 

0.25

2

2

2

2

0.5

2

2

2

2

1.00

2

2

2

2

2.5

2

2

2

2

5.0

1

2

2

2

10.00

1

2

1

2

corn oil

2

2

1

1

PRELIMINARY TEST

Dermal application

After 24, 48 and 72 h, no skin reaction was observed in the test animals in the concentration 2.5, 25 and 50% of the test substance. After 24 h very slight erythem and edema were observed in 2/4 animals being fully reversible after 48 h.

MAIN TEST

- Bodyweight changes and systemic effects:

No toxic effects were observed in the treated animals during the observation period.

One animal died within 24 h after the dermal induction. Possibly, the animal was too strong bandaged resulting in dyspnoea and death. Necropsy revealed a clear yellow liquid in the thorax and no further abnormal organ findings.

- Results of the intradermal induction:

One h after injection of FCA all test and control animals showed severe erythema and oedema. 11/20 test animals and 6/20 control animals showed injuries in the depth. 24 h later all animals showed severe erythema and oedema.

24 h after test substance injection all test animals showed slight erythema and oedema comparable to the control animals. Injection of test substance and FCA (1:1) induced moderate to strong erythema and moderate oedema. After 24 h strong erythema with deep injuries in 8/20 test animals and strong oedema were observed. Control animals injected with 50% FCA and corn oil showed 1 h after application well defined erythema and slight oedema. 24 h post application severe erythema with deep injuries (3/20) and strong oedema were observed.

 

- Results of the dermal induction:

Due to the use of SDS the shoulder region was reddened and swollen at Day 7. All test and control animals treated with FCA showed 49 h post application severe erythema with deep injuries and eschar formation and severe oedema. After 72 h the application sites of all animals were swollen and eschar formation was apparent. The animals treated with test substance in the vehicle showed 49 and 72 h post application scale formation.

Animals treated with the test substance in FCA showed 49 and 72 h after application severe erythema with injuries in the depth and strong oedema. The control animals showed comparable reaction at the application site.

 

- Results of the challenge application:

None of the test animals and none of the control animals showed a skin reaction 48 and 72 h post application.

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.

Data source

Materials and methods

Results and discussion

Applicant's summary and conclusion