o-Biphenyl-dimethylfluorenyl-amin

Endpoint summary

Description of key information

OECD 429: negative

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:

2016-07-18 to 2017-01-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

adopted 22 july 2010

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

updated 06 July 2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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: Envigo RMS B.V., Inc, Postbus 6174, NL - 5960, AD Horst
- Age at study initiation: 1st pre-test: 10-11 weeks, main study: 8 - 9 weeks
- Weight at study initiation: 1st pre-test: 20.3 & 20.7 g, main study: 17.3 - 21.7 g
- Housing: grouped per dose
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2°C
- Humidity (%): 45 – 65 %
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: day 1 To: day 5

Vehicle:

methyl ethyl ketone

Concentration:

1st pre-test: 10 and 25 %
main study: 5, 10, and 25 (w/w)

No. of animals per dose:

pre-test: 2
main study: 4 ( 3 treatment groups and on vehicle control group, total 16)

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: The highest test item concentration, which can be technically used was a 25% solution in DMF.
- Irritation / Systemic toxicity: From day 2 to 6, the animals showed an erythema of the ear skin (score 1, see Appendix 1 for details). No signs of excessive local skin irritation were present in both animals.
Thus, the test item in the main study was assayed at 5, 10, and 25%. The highest concentration tested was the highest level that could be achieved whilst avoiding systemic toxicity and excessive local skin irritation as confirmed in the pre-experiment.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: OECD 429
- Criteria used to consider a positive response: Stimulation index > 3

TREATMENT PREPARATION AND ADMINISTRATION:
Each test group of mice was treated by (epidermal) topical application to the dorsal surface of each ear with test item concentrations of 5, 10 and 25 in DMF. The application volume, 25 μL/ear/day, was spread over the entire dorsal surface (diameter approx. 8 mm) of each ear once daily for three consecutive days. A further group of mice (control animals) was treated with an equivalent volume of the relevant vehicle alone (control animals).
Five days after the first topical application (day 6) 250 μL of phosphate-buffered saline containing 3H-methyl thymidine were injected into each test and control mouse via the tail vein.
Approximately five hours after treatment with 3HTdR, all mice were euthanized by using CO2, draining lymph nodes were rapidly excised and pooled for each experimental group (8 nodes per group) and single cell suspensions (in phosphate buffered saline) of pooled lymph node cells were prepared.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

All calculations conducted on the DPM values were performed with a validated test script of “R”, a language and environment for statistical computing and graphics.

Positive control results:

Conc. SI
10% 5.51
25% 7.14
50% 7.31

Key result

Parameter:

SI

Value:

1.26

Test group / Remarks:

Test Group: 5 %

Key result

Parameter:

SI

Value:

1.09

Test group / Remarks:

Test Group: 10 %

Key result

Parameter:

SI

Value:

0.96

Test group / Remarks:

Test Group: 25 %

Cellular proliferation data / Observations:

Calculation of the EC3 value
The EC3 value could not be calculated, since all S.I.´s are below the threshold value of 3.

Viability / Mortality
No deaths occurred during the study period.

Clinical Signs
No signs of systemic toxicity were observed during the study period. On day 2 and 3 the animals showed an erythema of the ear skin

Body Weights
The body weight of the animals, recorded prior to the first application and prior to treatment with 3HTdR, was within the range commonly recorded for animals of this strain and age.

Calculation and Results of Individual Data

Test item concentration / [%]	Group	DPM	Calculation			Result
			DPM BG (a)	No of LN	DPM / LN (b)	S.I.
-	BG I	16	-	-	-	-
-	BG II	17	-	-	-	-
0	1	5467	5450.5	8	681.3	1.00
5	2	6884	6867.5	8	858.4	1.26
10	3	5943	5926.5	8	740.8	1.09
25	4	5257	5240.5	8	655.1	0.96

1 = Control Group
2-4 = Test Groups
BG - Background
DPM - disintegrations per minute
LN - Lymph Node
S.I. - Stimalation 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:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

The test item was not a skin sensitiser under the test conditions of this study.

Executive summary:

The informtion for this endpoint study record was obtained from an experimental study. The OECD GLP criteria were met and the methods applied are fully compliant with OECD TG 429. The test item was not a skin sensitiser under the test conditions of this study.

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

Based on the provided information there is no need for classification according to the EU Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.