Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Tested in this study according to OECD test guideline no. 429 and GLP conditions the test item was not a skin sensitizer.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 5 APR 2005 to 13 JUN 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD TG 429)
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Netherlands BV, Horst, The Netherlands
- Age at study initiation: 8-12 weeks
- Housing: Individually in Macrolon Type I cages with wire mesh top and granulated sooft wood bedding
- Diet: Pelleted standard diet (provided by Harlan Winkelmann GmbH, Borchen, Germany); ad libitum
- Water: Community tap water from Rossdorf, ad libitum
- Acclimation period: yes, but duration not mentioned

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
dimethyl sulphoxide
Concentration:
0, 5, 10, and 20% (W/v)
No. of animals per dose:
4 females per dose group
2 females in the pre-test
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: DMSO was selected because in any other vehicle test item concentrations above 20% could not be obtained
- 20% (w/v) was the highest technically applicable concentration
- Irritation: no systemic toxicity after single application of test item concentrations of 2.5, 5, 10 or 20% (w/v); irritating effects could not be assessed due to yellow staining at concentrations of 10 and 20% (w/v), but no ear swelling was observed
- Lymph node proliferation response: no data

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
1: exposure to at least one concentration of the test item resulted in an incorporation of 3H-methyl thymidine at least 3-fold or greater than that recorded in control mice as indicated by the stimulation index
2: data are compatible with a conventional dose response, although allowance must be made for either local toxicity or immunological suppression

TREATMENT PREPARATION AND ADMINISTRATION:
- individual preparation of weight volume dilutions using a magnetic stirrer as homogenizer
- test item preparations were made feshly before each dosing occasion
- 25 µl were spread over the entire dorsal surface of each ear lobe once daily for three consecutive days
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
- calculations of mean values and standard deviations for body weight
- the EC3 value was calculated according to the equation EC3 = (a-c) [(3-d)/(b-d)] + c; where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining Iymph node cell proliferative activity; (a, b) and (c, d) are respectively the co-ordinates of the two pair of data lying immediately above and below the value of 3 on the local lymph node assay dose response Plot.
Positive control results:
Stimulation indices of 2.29, 3.21 and 8.44 were determined with the positive control item at concentrations of 5%, 10% and 25% (w/v), respectively, in acetone:olive oil, 4:1 (v/v). An EC3 value was calculated (= 8.9% (w/v)).
Parameter:
SI
Remarks on result:
other: Stimulation indices (SI) were all below 3. The following SI were calculated: 5% test item: 1.87 10% test item: 1.79 20% test item: 2.35
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Background of measurement: 16.9 - 28.9; mean: 22.9 (was taken for calculations) control group: 4397.9 5% test item: 8187.8 10% test item: 7868.1 20% test item: 10291.3

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. Calculation of the EC3 value was not performed because no test concentration produced a SI of 3 or higher.

Test item
concentration
% (w/v)

Group

Measurement
DPM

Calculation

Result

DPM-BGa)

number of
lymph nodes

DPM per
lymph nodeb)

S.I.

---

BG I

28.9

---

---

---

---

---

BG II

16.9

---

---

---

---

---

CG 1

4397.9

4375.0

8

546.9

 

5

TG 2

8187.8

8164.9

8

1020.6

1.87

10

TG 3

7868.1

7845.2

8

980.7

1.79

20

TG 4

10291.3

10268.4

8

1283.6

2.35

BG = Background (1 ml 5% trichloroacetic acid) in duplicate

CG = Control Group

TG = Test Group

S.I. = Stimulation Index

a)       = The mean value was taken from the figures BG I and BG II

b)       = Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Tested in this study according to OECD test guideline no. 429 and GLP conditions the test item was not a skin sensitizer.
Executive summary:

In the study the test item dissolved in DMSO was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 5, 10, and 20 %.

No cases of mortality were observed.No symptoms of systemic findings were observed during the study period. Due to the intense yellow colour of the test item at 10 and 20 % (w/v) local irritation reactions such asear redness could not be detected. No swelling of the ears was observed at any of the animals.

In this study Stimulation Indices (S.I.) of 1.87, 1.79, and 2.35 were determined with the test item at concentrations of 5, 10, and 20 % (w/v) in DMSO, respectively.

The test item was found to be not a skin sensitiser in this assay.

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:
skin sensitisation: in chemico
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:
skin sensitisation: in vivo (LLNA)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From 5 APR 2005 to 13 JUN 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD TG 429)
Justification for type of information:
See read across justification document in chapter 13
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Netherlands BV, Horst, The Netherlands
- Age at study initiation: 8-12 weeks
- Housing: Individually in Macrolon Type I cages with wire mesh top and granulated sooft wood bedding
- Diet: Pelleted standard diet (provided by Harlan Winkelmann GmbH, Borchen, Germany); ad libitum
- Water: Community tap water from Rossdorf, ad libitum
- Acclimation period: yes, but duration not mentioned

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
dimethyl sulphoxide
Concentration:
0, 5, 10, and 20% (W/v)
No. of animals per dose:
4 females per dose group
2 females in the pre-test
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: DMSO was selected because in any other vehicle test item concentrations above 20% could not be obtained
- 20% (w/v) was the highest technically applicable concentration
- Irritation: no systemic toxicity after single application of test item concentrations of 2.5, 5, 10 or 20% (w/v); irritating effects could not be assessed due to yellow staining at concentrations of 10 and 20% (w/v), but no ear swelling was observed
- Lymph node proliferation response: no data

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
1: exposure to at least one concentration of the test item resulted in an incorporation of 3H-methyl thymidine at least 3-fold or greater than that recorded in control mice as indicated by the stimulation index
2: data are compatible with a conventional dose response, although allowance must be made for either local toxicity or immunological suppression

TREATMENT PREPARATION AND ADMINISTRATION:
- individual preparation of weight volume dilutions using a magnetic stirrer as homogenizer
- test item preparations were made feshly before each dosing occasion
- 25 µl were spread over the entire dorsal surface of each ear lobe once daily for three consecutive days
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
- calculations of mean values and standard deviations for body weight
- the EC3 value was calculated according to the equation EC3 = (a-c) [(3-d)/(b-d)] + c; where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining Iymph node cell proliferative activity; (a, b) and (c, d) are respectively the co-ordinates of the two pair of data lying immediately above and below the value of 3 on the local lymph node assay dose response Plot.
Positive control results:
Stimulation indices of 2.29, 3.21 and 8.44 were determined with the positive control item at concentrations of 5%, 10% and 25% (w/v), respectively, in acetone:olive oil, 4:1 (v/v). An EC3 value was calculated (= 8.9% (w/v)).
Parameter:
SI
Remarks on result:
other: Stimulation indices (SI) were all below 3. The following SI were calculated: 5% test item: 1.87 10% test item: 1.79 20% test item: 2.35
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Background of measurement: 16.9 - 28.9; mean: 22.9 (was taken for calculations) control group: 4397.9 5% test item: 8187.8 10% test item: 7868.1 20% test item: 10291.3

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. Calculation of the EC3 value was not performed because no test concentration produced a SI of 3 or higher.

Test item
concentration
% (w/v)

Group

Measurement
DPM

Calculation

Result

DPM-BGa)

number of
lymph nodes

DPM per
lymph nodeb)

S.I.

---

BG I

28.9

---

---

---

---

---

BG II

16.9

---

---

---

---

---

CG 1

4397.9

4375.0

8

546.9

 

5

TG 2

8187.8

8164.9

8

1020.6

1.87

10

TG 3

7868.1

7845.2

8

980.7

1.79

20

TG 4

10291.3

10268.4

8

1283.6

2.35

BG = Background (1 ml 5% trichloroacetic acid) in duplicate

CG = Control Group

TG = Test Group

S.I. = Stimulation Index

a)       = The mean value was taken from the figures BG I and BG II

b)       = Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Tested in this study according to OECD test guideline no. 429 and GLP conditions the test item was not a skin sensitizer.
Executive summary:

In the study the test item dissolved in DMSO was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 5, 10, and 20 %.

No cases of mortality were observed.No symptoms of systemic findings were observed during the study period. Due to the intense yellow colour of the test item at 10 and 20 % (w/v) local irritation reactions such asear redness could not be detected. No swelling of the ears was observed at any of the animals.

In this study Stimulation Indices (S.I.) of 1.87, 1.79, and 2.35 were determined with the test item at concentrations of 5, 10, and 20 % (w/v) in DMSO, respectively.

The test item was found to be not a skin sensitiser in this assay.

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

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

No classification

Tested in this study according to OECD test guideline no. 429 and GLP conditions the test item was not a skin sensitizer.