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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Reference:
Composition 0
Principles of method if other than guideline:
The modified Local Lymph Node Assay (IMDS) was performed on 24 female NMRI mice of the strain Crl:NMRI BR (6 animals/test item group and 6 control animals) to determine if there is any specific (sensitizing) or non-specific (irritant) stimulating potential of the test item Macrolex Blau 3R.
The modifications refer to the measurement of cell proliferation by cell counting instead of radioactive labeling. In addition, the acute inflammatory skin reaction is determined to discriminate specific from non-specific activation of immune competent cells in the draining lymph nodes.
GLP compliance:
yes
Type of study:
mouse local lymphnode assay (LLNA)
Test material information:
Composition 1
Species:
mouse
Strain:
NMRI
Sex:
female
Vehicle:
dimethylformamide
Concentration:
Group 1 Vehicle (DMF)
Group 2 2% Macrolex Blau 3R (in DMF)
Group 3 10% Macrolex Blau 3R (in DMF)
Group 4 50% Macrolex Blau 3R (in DMF)
Group 5 40% Alpha Hexyl Cinnamic Aldehyde (in DMF)
No. of animals per dose:
6 animals/test item group and 6 control animals
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
A modified Local Lymph Node Assay (IMDS) was carried out in mice. The modifications refer to the measurement of cell proliferation by cell counting instead of radioactive labeling. Tabular summary of the LLNAlIMDS results 1. Direct LLNA (NMRI mice, female, 6 animals/group) Groups Weight index Cell count index (index of mean +/-SD in 0/0) Gr. 1 1.00 +/-25.83 1.00 +/-39.31 Gr. 2 0.84 +/-23.33 0.94 +/-27.50 Gr. 3 0.81 +/-33.06 0.97 +/-38.44 Gr. 4 1.06 +/-23.52 1.24 +/-35.52 Gr. 5 1.58s +/-24.81 1.90s +/-24.42 2. Ear swelling (NMRI mice, female, 6 animals/group, in 0.01 mm) Groups day 1 day 4 Index day 4 (mean +/-SD in %) Gr. 1 17.33 +/-2.84 17.75 +/-2.55 1.00 Gr. 2 17.17 +/-2.27 17.58 +/-3.80 0.99 Gr. 3 17.17 +/-3.36 17.75 +/-4.88 1.00 Gr. 4 17.25 +/-2.62 17.75 +/-2.55 1.00 Gr. 5 17.00 +/-3.55 28.08s +/-17.54 1.58 3. Ear weight (NMRI mice, female, 6 animals/group, in mg per 8 mm diameter punch) Groups day 4 Index day 4 (mean +/-SO in %) Gr. 1 12.01 +/-5.49 1.00 Gr. 2 12.09 +/-3.45 1.01 Gr. 3 12.38 +/-6.32 1.03 Gr. 4 13.30 +/-11.65 1.11 Gr. 5 18.12s +/-11.42 1.51 s = statistically significant increase (p< 0.05) Group 1 Vehicle (DMF) Group 2 2% Macrolex Blau 3R (in DMF) Group 3 10% Macrolex Blau 3R (in DMF) Group 4 50% Macrolex Blau 3R (in DMF) Group 5 40% Alpha Hexyl Cinnamic Aldehyde (in DMF)
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: not applicable

Based on results obtained in validation studies and general experiences with this test system groups of mice were treated with vehicle, 2 %, 10 % or 50 % Macrolex Blau 3R in DMF.

The NMRI mice did not show an increase in the stimulation indices for cell counts or for weights of the draining lymph nodes after application of the test item Macrolex Blau 3R.

The “positive level”, which is 1.4 for the cell count index, was never reached or exceeded in any dose group.

The “positive level” of ear swelling, which is 2x10E-2 mm increase, i.e. about 10 % of the control values, has not been reached or exceeded in any dose group.

No substance specific effects were determined for ear weights either.

After treatment with Alpha Hexyl Cinnamic Aldehyde (group 5) the NMRI mice showed clear increases in the weights of the draining lymph nodes and in the stimulation indices for cell counts compared to control animals, which are of statistical significance. The “positive level”, which is 1.4 for cell count indices, has clearly been exceeded.

The “positive level” of ear swelling, which is 2x10E-2 mm increase, i.e. about 10 % of the control values, has been exceeded in the positive control group. This increase is of statistical significance. A significant increase compared to vehicle treated animals regarding ear weights was detected, too.

It has to be clarified that the “positive levels” mentioned above are exclusively defined for the NMRI outbreed mice used for this study. Such positive limits have to be calculated for each strain of mice individually.

Interpretation of results:
not sensitising
Remarks:
Migrated information
Executive summary:

A modified Local Lymph Node Assay (IMDS) was performed on 24 female NMRI mice of the strain HsdWin:NMRI (6 animals/test item group and 6 control animals) to determine if there is any specific (sensitizing) or non-specific (irritant) stimulating potential of the test item Macrolex Blau 3R.

A concurrent control of 6 animals treated with Alpha Hexyl Cinnamic Aldehyde was included.

The study was conducted according to OECD Guidelines No. 429 and No. 406,

EC Guideline 2004/73/EC (29th Adaptation of Guideline 67/548/EEC, B.42)/Health Effects Test Guideline and OPPTS 870.2600 (EPA) with the following test item concentrations:

Test item: 0 % (vehicle control), 2 %, 10 % and 50 %.

Positive control: 40 % Alpha Hexyl Cinnamic Aldehyde

The test item was formulated in dimethylformamide (DMF) to yield a suspension.

The positive control was formulated in dimethylformamide (DMF) to yield a solution.

Compared to vehicle-treated animals, none of the parameters measured in the substance-treated groups, i.e. cell counts and weights of the draining lymph nodes, ear weights and ear swelling, reached or exceeded the "positive levels" defined for this assay. These results show that there is no indication for a skin sensitizing effect after administration of a concentration up to and including 50 % Macrolex Blau 3R in this test system.

In conclusion, these results show that the test item Macrolex Blau 3R has no sensitizing potential in mice after dermal application of up to and including a 50 % concentration. No indication for a non-specific (irritant) activation was detected, too. Therefore, the concentration of 50 % turned out to be the NOEL for the parameters investigated in this study with respect to skin sensitization.

These results are verified by the comparison with the results of the positive control group (Alpha Hexyl Cinnamic Aldehyde).

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

Macrolex Blau 3R has no sensitizing potential in mice after dermal application of up to and including a 50 % concentration (highest applied dose). No indication for a non-specific (irritant) activation was detected, too. Therefore, the concentration of 50 % turned out to be the NOEL for the parameters investigated in this study with respect to skin sensitization.


Migrated from Short description of key information:
A modified Local Lymph Node Assay (IMDS) was performed on 24 female NMRI mice of the strain HsdWinin:NMRI (6 animals/test item group and 6 control animals) to determine if there is any specific (sensitizing) or non-specific (irritant) stimulating potential of the test item 1,4-bis[(2-ethyl-6-methylphenyl)amino]anthraquinone.

Justification for selection of skin sensitisation endpoint:
key study used

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

No data


Migrated from Short description of key information:
No data

Justification for classification or non-classification

In the LLNA 1,4-bis[(2-ethyl-6-methylphenyl)amino]anthraquinone has no sensitizing potential in mice after dermal application of up to and including a 50 % concentration. Therefore a classificationis not justifed.