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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From January 21, 2008 to May 07, 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: approx. 10 weeks old
- Housing: Individual in labeled Macrolon cages (MI type; height 12.5 cm)
- Acclimation period: at least 5 days
- Temperature (°C): 21.0 ± 3.0 °C
- Humidity (%): 30-70 %
- Photoperiod: 12-hrs dark / 12-hrs light

Study design: in vivo (LLNA)

Vehicle:
propylene glycol
Concentration:
For preliminary study: 25% and 50%
For main study: 0% (vehicle), 10%, 25% and 50%
No. of animals per dose:
For preliminary study: one per dose
For main study: five per dose
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
The EC3 of hexyl cinnamic aldehyde (HCA) was 14.1%.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Value:
2.2
Test group / Remarks:
10% test substance
Parameter:
SI
Value:
7.4
Test group / Remarks:
25% test substance
Parameter:
SI
Value:
6.8
Test group / Remarks:
50% test substance

Any other information on results incl. tables

In preliminary study, the results indicated that no irritation was observed in any of the animals examined.

The results in main study as below:

 

Table 1. Skin reactions, body weights and relative size auricular lymph nodes

group

% test substance1

an2

Day1

Day3

Day6

bw (g)3

Skin reactions dorsal surface ear

bw (g)3

Size nodes4

left

right

left

right

erythema

oedema

erythema

oedema

1

0%

(vehicle)

1

23

0

0

0

0

21

n

n

2

23

0

0

0

0

22

n

n

3

21

0

0

0

0

20

n

n

4

23

0

0

0

0

22

n

n

5

20

0

0

0

0

21

n

n

2

10%

6

22

G

0

G

0

22

n

n

7

21

G

0

G

0

20

n

n

8

20

G

0

G

0

20

n

n

9

22

G

0

G

0

22

n

n

10

24

G

0

G

0

24

n

n

3

25%

11

22

G

0

G

0

21

+

+

12

21

G

0

G

0

21

+

+

13

21

G

0

G

0

21

+

+

14

23

G

0

G

0

21

+

+

15

20

G

0

G

0

20

+

+

4

50%

16

21

G

0

G

0

22

+

+

17

21

G

0

G

0

19

+

+

18

22

G

0

G

0

20

+

+

19

23

G

0

G

0

21

+

+

20

23

G

0

G

0

22

n

n

1 Vehicle: Propylene glycol

2 Animal number

3 Body weight (g)

4 Relative size auricular lymph nodes (-, -- or ---: degree of reduction, +, ++ or +++: degree of enlargement, n: considered to be normal)

G No scoring possible due to red staining of the ears

 

Table 2. Radioactivity measurements (individual animals)

group

% test substance1

animal

DPM / animal

1

0%

(vehicle)

1

92

2

286

3

336

4

174

5

112

2

10%

6

47

7

668

8

863

9

863

10

95

3

25%

11

2093

12

1341

13

3214

14

543

15

1230

4

50%

16

1240

17

1411

18

2669

19

2074

20

378

1       Vehicle: Propylene glycol

2       Value rejected and not used for interpretation (outside historical range for commonly used vehicles)

 

Table 3. Disintegrations Per Minute (DPM) and Stimulation Index (SI)

Group

% test substance1

mean

DPM±SEM

SI±SEM

2

10%

507 ± 182

2.2 ± 0.9

3

25%

1684 ± 455

7.4 ± 2.6

4

50%

1554 ± 388

6.8 ± 2.3

1

0% (vehicle)

227 ± 51

1.0 ± 0.3

1 Vehicle: Propylene glycol

Applicant's summary and conclusion

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
According to OECD 429 test method, the SI at 25% and 50% are 7.4 and 6.8 respectively. The EC3 value of Everzol Red CDN Crude was 12.3%. Therefore, Everzol Red CDN Crude was category 1 based on GHS criteria.
Executive summary:

This test using the procedures outlined in the NOTOX Study Plan for 487479. The results of this OECD 429 test for Everzol Red CDN Crude show that test reliability criteria was met. Based on the results of preliminary study, a 50% concentration was selected as the highest concentration that could be used in the main study. In the main study, three group of five experimental animals were treated with test substance concentrations of 10%, 25% or 50% on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Propylene glycol). Three days after the last exposure, all animals were injected with H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised. After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of DPM and SI was subsequently calculated for each group.

The results indicated that no skin reactions were observed in any of the animals examined and no scoring for erythema was possible due to red staining by the test substance. Enlarged nodes were found in higher dose groups. Body weights and bodyweight gain of experimental animals remained in the same range as control over the study period. No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Mean DPM/animal value for the experimental groups treated with concentrations 10, 25 and 50 % Everzol Red CDN Crude were 507, 1684 and 1554 respectively. On the basis of the test results given above, the SI at 25% and 50% are 7.4 and 6.8 respectively. The EC3 value of Everzol Red CDN Crude was 12.3%.