Registration Dossier

Administrative data

Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
8 February 2010 - 22 March 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with minor deviations, which had no impact on the integrity of the study.
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
Temporary deviations from the minimum level of relative humidity occurred.
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
Temporary deviations from the minimum level of relative humidity occurred.
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
yes
Remarks:
Temporary deviations from the minimum level of relative humidity occurred.
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
other: CBA/J
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS- Source:Charles River France, L’Arbresle Cedex, France.- Age at study initiation: Young adult animals (approx. 9 weeks old)- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean.- Housing: Macrolon cages (MIII type; height 18 cm).- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).- Water (e.g. ad libitum): Free access to tap water.- Acclimation period: at least 5 days before the start of treatment under laboratory conditionsENVIRONMENTAL CONDITIONS- Temperature (°C): 21.0 ± 3. 0ºC (actual range: 18.5 -22. 7ºC)- Humidity (%): 40-70% (actual range: 39 - 65%)- Air changes (per hr): 15- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.
Vehicle:
dimethylformamide
Concentration:
0%, 10%, 25% and 50%
No. of animals per dose:
5 animals per group
Details on study design:
RANGE FINDING TESTS:- Compound solubility:- Irritation:- Lymph node proliferation response:MAIN STUDYANIMAL ASSIGNMENT AND TREATMENT- Name of test method:- Criteria used to consider a positive response: DPM values are presented for each animal and for each dose group. A Stimulation Index (SI) is calculated for each group. The SI is the ratio of the DPM/group compared to DPM/vehicle control group.If the results indicate a SI ≥ 3, the test substance may be regarded as a skin sensitizer, based on the test guideline and recommendations done by ICCVAM (Reference 1). The results were evaluated according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations (2007) and the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labeling and packaging of substances and mixtures. The EC3 value (the estimated test substance concentration that will give a SI =3) was determined, using linear interpolationTREATMENT PREPARATION AND ADMINISTRATION:Grading Irritation Reactions:Erythema and eschar formation:No erythema ..............................................................................……………………………………….. ......... 0Slight erythema (barely perceptible) ....................................................……………………………… ............ 1Well-defined erythema ...................................................................…………………………………… ........... 2Severe erythema (beef redness) to slight eschar formation (injuries in depth) .........……………… ............ 3Oedema formation:No oedema .................................................................................……………………….……………… ......... 0Slight oedema (barely perceptible) ......................................................……………………………… ............ 1Moderate oedema....................................…………………… ....................................................................... 2Severe oedema… ....................................................................................................................................... 3
Positive control substance(s):
other: dimethyl formamide
Statistics:
see below
Positive control results:
See Table 1 below.
Parameter:
SI
Remarks on result:
other: 1.7 for the substance concentration 10%2.6 for the substance concentration 25%4.7 for the substance concentration 50%
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: 481 DPM at 10% concentration of Modified Small Vinyl Ester 728 DPM at 25% concentration of Modified Small Vinyl Ester 1329 DPM at 50% concentration of Modified Small Vinyl Ester vehicle control group: 281 DPM

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

Group

test

substance1

(% w/w)

an2

Day 1

Day 3

Skin reactions dorsal surface ear

Day 6

bw

left

right

bw

size nodes4

(g)3

erythema

oedema

erythema

oedema

(g)3

Left 

right

1

0%

vehicle

1

19

0

0

0

0

20

n

n

2

20

0

0

0

0

19

n

n

3

19

0

0

0

0

20

n

n

4

18

0

0

0

0

18

+

n

5

18

0

0

0

0

18

n

n

2

10%

6

20

1

0

1

0

18

n

n

7

18

1

0

1

0

18

n

n

8

18

1

0

1

0

18

n

n

9

19

1

0

1

0

19

n

n

10

18

1

0

1

0

18

n

n

3

25%

11

19

15

0

15

0

19

n

n

12

19

15

0

15

0

19

n

n

13

20

15

0

15

0

20

n

n

14

20

15

0

15

0

21

n

n

15

19

15

0

15

0

20

n

n

4

50%

16

19

15

0

15

0

18

n

+

17

19

15

0

15

0

19

n

n

18

21

15

0

15

0

22

+

+

19

21

15

0

15

0

20

n

+

20

18

15

0

15

0

18

+

+

 

   1Vehicle: N,N-dimethylformamide.

2Animal number.

3Body weight (grams).

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

5Bald area on head and ears.

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Remarks:
Migrated information
Conclusions:
Assessment of Contact Hypersensitivity to Modified Small Vinyl Ester was tested in the murine local lymph node assay (LLN A) in accordance the guidelines (OECD 429). The results showed that Modified Small Vinyl Ester elicited a SI> or equal to 3. The data showed a dose-response and an EC3 value of 29.8% was calculated. Thus, Modified Small Vinyl Ester was sensitizing in the murine local lymph node assay.
Executive summary:

Assessment of Contact Hypersensitivity to Modified Small Vinyl Ester in the Mouse (Local Lymph Node Assay). The study was carried out in accordance with the guidelines (e.g. OECD 429). The concentrations of Modified Small Vinyl Ester selected for the main study were based on the results of a preliminary study. In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 10, 25 or 50% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (N,N-dimethylformamide). Three days after the last exposure, all animals were injected with 3H-methylthymidine and five hours later the draining (auricular) lymph nodes were excised. After precipitation of the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of disintegrations per minute (DPM) and a stimulation index (SI) was subsequently calculated for each group.

.

Slight irritation of the ears was shown by all animals treated at 10, 25% and 50%. The animals at the 25 and 50% dose groups showed also bald area on the head and ears. These skin reactions were considered not to have a toxicologically significant effect on the activity of the nodes. No oedema was observed in any of the animals examined. The auricular lymph nodes of most control animals and all animals at 10 and 25% were considered normal in size, except for one node of one control animal, which was increased in size. At the highest dose group the majority of animals showed increased size of the lymph nodes. No macroscopic abnormalities of the surrounding area were noted in any of the animals. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. The slight body weight loss, noted in some animals, was considered not toxicologically significant.

Mean DPM/animal values for the experimental groups treated with Modified Small Vinyl Ester concentrations 10, 25 and 50% were 481, 728 and 1329 DPM respectively. The mean DPM/animal value for the vehicle control group was 281 DPM. The SI values calculated for test concentrations 10, 25 and 50% were 1.7, 2.6 and 4.7, respectively. The results showed that Modified Small Vinyl Ester elicited a SI ≥ 3. The data showed dose-response and an EC3 value (the estimated test substance concentration that will give a SI =3) of 29.8% was calculated. Based on these results, Modified Small Vinyl Ester was sensitizing in the muirne local lymph node assay.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Modified Small Vinyl Ester was tested in a murine Local Lymph Node Assay. The study was carried out in accordance with the guidelines (e.g. OECD 429). The concentrations of Modified Small Vinyl Ester selected for the main study were based on the results of a preliminary study. In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 10, 25 or 50% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (N,N-dimethylformamide). Three days after the last exposure, all animals were injected with 3H-methylthymidine and five hours later the draining (auricular) lymph nodes were excised. After precipitation of the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of disintegrations per minute (DPM) and a stimulation index (SI) was subsequently calculated for each group. The SI values calculated for test concentrations 10, 25 and 50% were 1.7, 2.6 and 4.7, respectively. The results show that Modified Small Vinyl Ester elicited a SI ≥ 3. The data showed dose-response and an EC3 value (the estimated test substance concentration that will give a SI =3) of 29.8% was calculated. Based on these results, Modified Small Vinyl Ester was sensitizing in the murine local lymph node assay.

Assessment of Contact Hypersensitivity to Modified Small Vinyl Ester in the Albino Guinea Pig (Maximization Test) was tested in a guideline study (OECD 406). The validity of the test method was controlled with a concurrent study using alpha-hexylcinnamic aldehyde. Modified Small Vinyl Ester concentrations selected for the main study were based on the results of a preliminary study. In the main study, ten experimental animals were intradermally injected with a 10% concentration and epidermally exposed to a 100% concentration. Five control animals were similarly treated, but with vehicle alone (propylene glycol). Approximately 24 hours before the epidermal induction exposure all animals were treated with 10% SDS. Two weeks after the epidermal application all animals were challenged with a 100% Modified Small Vinyl Ester and the vehicle.The signs of necrosis seen in the control animals after the intradermal injection with vehicle only were consistent with the results in the preliminary irritation study after injection of propylene glycol. The reactions noted in the experimental and control animals after the epidermal induction exposure were considered to be enhanced by the SDS treatment. Skin reactions of grade 1 were observed in 5 experimental animals in response to the 100% Modified Small Vinyl Ester. No skin reactions were evident in the vehicle control animals. No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Body weight and body weight gain of experimental animals remained in the same range as controls over the study period. The skin reactions observed in response to a 100% Modified Small Vinyl Ester in 5/10 experimental animals in the challenge phase were considered indicative of sensitization, based on the absence of any response in the control animals. These results indicate a sensitization rate of 50 per cent.


Migrated from Short description of key information:
In a murine Local lymph node assay (LLNA) Modified Small Vinyl Ester induced dose-related skin reactions in the challenge phase and elicited a stimulation index (SI) larger than 3. An EC3 value of 29.8% was calculated. Thus, Modified Small Vinyl Ester was sensitising in the LLNA. In a guinea pig maximisation test Modified Small Vinyl Ester induced a sensitization rate of 50% (5/10 animals) after challenge with 100% Modified Small Vinyl Ester. Thus, Modified Small Vinyl Ester was also sensitizing in the guinea pig maximisation test.

Justification for selection of skin sensitisation endpoint:
Study conducted using the more modern LLNA protocol

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Modified Small Vinyl Ester was sensititizing both in a murine local lymph node assay and in a Guinea Pig Maximization Test. On this basis the substance is classified for skin sensitisation (Skin sensitiser, category 1B).