Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2010

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)
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Test material form:
liquid: viscous
Details on test material:
- Manufacturers identification: Novares LA 300
- Substance type: organic
- Test material is 'Oligomerisation and alkylation reaction products of 2-phenylpropene and phenol' (OAPP), EC list number 700-960-7 (assigned manually to validated substances from inquiries by ECHA). Originally the substance phenol, methylstyrenated, CAS No. 68512-30-1, EC No. 270-966-8 was submitted for registration. Subsequent to substance validation, the identity of the substance was changed by ECHA.
- for additional information see respective study record
Specific details on test material used for the study:
- Name of test material (as cited in study report): Novares LA 300 (phenol, methylstyrenated)
- Lot/batch No.: 28166
- Composition of test material: composition is specified in IUCLID Sect. 13 - Assessment reports under Certificate of Analysis_Novares LA 300_phenol, methylstyrenated
- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light

In vivo test system

Test animals

Species:
mouse
Strain:
Balb/c
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeding farm VELAZ s.r.o., Koleč u Kladna, Czech Republic
- Age at study initiation: 8 - 10 wks
- Weight at study initiation: 17 - 19 g
- Housing: macrolon cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +- 3
- Humidity (%): 30 – 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Study design: in vivo (LLNA)

Vehicle:
other: DAE 433
Concentration:
mixture of 40% dimethylacetamide, 30% acetone and 30% ethanol
No. of animals per dose:
5
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: dispersion
- Irritation: no
- Lymph node proliferation response: not examined


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response:
Stimulation index (SI) is ≥ 3
Response increases in dose-related manner
Criteria for irritation potential: individual and mean ear weight


TREATMENT PREPARATION AND ADMINISTRATION:
Days 1-3: 25 µL test suspension in DAE 433 , 1x on day 1, 2, and 3
Days 4 and 5: no treatment
Day 6: Injection of 250 μL of phosphate-buffered saline (PBS) containing 7.5 x105 Bq (c. 21 µCi) of 3H-methyl thymidine
into all test and control mice via the tail vein. Five hours later, the animals were killed.



Positive control substance(s):
other: DNCB (dinitrochlorbenzene, 0.5% (w/v) solution)
Statistics:
Non-parametric Kruskal-Wallis test, and then the non-parametric two-group Mann-Whitney rank test (probability level 0.05) was applied to all two-group comparisons [using software Statgraphic ® Centurion (version XV, USA)].

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Key result
Parameter:
EC3
Value:
3.63
Parameter:
SI
Value:
2.36
Test group / Remarks:
0.3% test substance in vehicle; 5 animals
Parameter:
SI
Value:
2.68
Test group / Remarks:
3% test substance in vehicle; 5 animals
Parameter:
SI
Value:
16.35
Test group / Remarks:
30% test substance in vehicle; 5 animals

Any other information on results incl. tables

Table 1: Individual and mean radioactivity in cell suspensions (Report Table 4)

Activity (DPM)

Animal No.

NC

(1-5)

PC

(6-10)

30%

(11-15)

3%

(16-20)

0.3%

(21-25)

1

550.98

11374.69

9354.53

1425.77

1232.24

2

485.39

10401.94

9984.31

1475.60

1484.48

3

568.93

10347.82

11634.71

1367.08

1128.80

4

526.30

8837.38

9107.79

1301.28

1222.94

5

596.74

9493.34

4517.77

1748.67

1091.42

 Group

mean

545.67

10091.03

8919.82

1463.68

1231.98

SI

1.00

18.49

16.35

2.68

2.26

NC = vehicle control; PC = positive control

Table 2: Individual and average ear weights (Report Table 5)

Weight of ear biopsies (milligrams)

Animal No.

NC

(1-6)

PC

(7-12)

30%

(13-18)

3%

(19-24)

0.3%

(25-30)

1

24.5

27.4

32.7

25.0

21.8

2

25.2

27.5

28.4

23.0

23.1

3

23.8

26.5

34.2

21.4

21.4

 4

23.4

26.7

29.4

21.5

21.0

5

24.7

27.2

41.2

21.7

25.0

 Group mean

24.32

27.06*

33.18*

22.52

22.46

SD

0.72

0.44

5.07

1.53

1.62

NC = vehicle control; PC = positive control

* statistically significant with p =< 0.05, Mann-Whitney test

Table 3:  Summary table (Report Table 6)

Group

Radioisotope incorporation

in lymph nodes

Ear weight

Mean DPM

SI

Mean (mg)

NC

545.67

1.0

24.32

PC

10091.03

18.49+

27.06

30%

8919.82

16.35+

33.18

3%

1463.68

2.68

22.52

0.3%

1231.98

2.26

22.46

Bold figures with + = values >= 3

Applicant's summary and conclusion

Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria
Conclusions:
In this LLNA according to OECD TG 429, an EC3 of 3.63 was determined for the substance oligomerisation and alkylation reaction products of 1-phenylpropene and phenol. Resulting EU/GHS classification is skin sensitising Cat. 1B (EC3 > 2).