Reaction mass of benzyldodecylbis(2-hydroxypropyl)ammonium chloride and benzylbis(2-hydroxypropyl)tetradecylammonium chloride

Endpoint summary

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: guideline study

Qualifier:

according to guideline

Guideline:

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

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 Katalysator W AZ 5596-B.

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

NMRI

Sex:

female

Vehicle:

dimethylformamide

Concentration:

Group 1 Vehicle (DMF)
Group 2 2% Katalysator W AZ 5596-B (in DMF)
Group 3 10% Katalysator W AZ 5596-B (in DMF)
Group 4 50% Katalysator W AZ 5596-B (in DMF)
Group 5 30% 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. 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, as also recommend in the update of OECD TG 429. 1. Direct LLNA (NMRI mice, female, 6 animalsl group) Groups Weight index Cell count index (index of mean +/-SD in %) Gr.l 1.00 +/-27.79 1.00 +/-32.01 Gr.2 1.88 +/-11.46 1.78 +/-14.11 Gr.3 1.68 +/-39.95 1.84 +/-35.98 Gr.4 1.69 +/-43.54 1.08 +/-65.67 Gr. 5 2.01 +/-29.30 2.39 +/-36.11 Ear swelling (NMRI mice, female, 6 animals/group, in 0.01 mm) Groups day 1 day4 Index day4 (mean +/- SD in %) Gr. 1 17.25 +/- 2.62 17.67 +/- 2.79 1.00 Gr.2 17.25 +/- 3.60 18.67 +/- 6.98 1.06 Gr. 3 17.33 +/- 3.76 21.42 +/- 15.61 1.21 Gr. 4 17.33 +/- 5.12 36.92 +/- 16.31 2.09 Gr. 5 17.50 +/- 3.85 26.75 +/- 25.01 1.51 Ear weight (NMRI mice, female, 6 animals/group, in mg per 8 mm diameter punch) Groups day4 Index day4 (mean +/- SD in %) Gr.1 12.32 +/- 8.82 1.00 Gr.2 14.77 +/- 6.56 1.20 Gr.3 18.47 +/- 8.63 1.50 Gr.4 34.57 +/- 9.64 2.81 Gr.5 19.53 +/- 17.97 1.59 Group 1 Vehicle (DMF) Group 2 2% Katalysator W AZ 5596-B (in DMF) Group 3 10% Katalysator W AZ 5596-B (in DMF) Group 4 50% Katalysator W AZ 5596-B (in DMF) Group 5 30% Alpha Hexyl Cinnamic Aldehyde (in DMF)

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: not applicable

A sensitizing potential can be assumed from the increases in cell proliferation in the draining lymph nodes. On the basis of our experiences using this method the "positive level" had been set to an increase in cell count index by 0.4 (i.e. index >1.4), which has been exceeded in the low and the mid dose group. Differentiation indices (DI), which is the quotient of the relative lymph node reaction divided by the relative acute skin reaction were > 1 for the low and the mid concentration tested, i. e. 3.25, and 1.00, respectively . These DI values do point to a sensitizing potential ofthe test item.

The "positive level" of ear swelling which is 2x 10-2 rnm increase, i.e. about 10% of the control values, has been exceeded in the mid and in the high dose group. An increase in this parameter does point to an acute irritant (inflammatory) response. Due to

the strong irritant property of the test item at these concentrations it is not quite clear if the cell proliferation is partly induced by this non-specific (inflammatory) reaction. On the other hand effects measured at the low concentration (2%) clearly point to a sensitizing property of the test item.

Although it is not possible to calculate an exact EC value from the data obtained, it can be assumed that the EC value is in any case below 2%. In accordance with the classification proposed in the Technical Report No. 78 of the ECETOC this value corresponds to at least a moderate skin sensitizer.

Interpretation of results:

sensitising

Remarks:

Migrated information

Executive summary:

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 Katalysator WAZ 5596-B.

This study was carried out with NMRI mice from commercial breeder Charles River Germany, which is not the normally used breeder of the lab. Therefore, 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 2004173IEC (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 % (vehic1e control), 2 %, 10 % and 50 %. Positive control: 30 % Alpha Hexyl Cinnamic Aldehyde. The test item was formulated in dimethylformarnide (DMF) to yield a solution.

Compared to vehicle treated animals there was a clear increase regarding the weights of the draining lymph nodes, which is of statistical significance in the low dose group and a clear increase in the cell counts, which is of statistical significance in the low and the rnid dose group. The "positive level" of index 1.4 for the cell counts has been exceeded in the low and the mid dose group.

The "positive level' of ear swelling which is 2x 10-2 mm increase, i.e. about 10% of the control values, has been exceeded in the rnid and the high dose group. These changes are of statistical significance. A significant increase compared to vehicle treated animals regarding ear weights was detected in all dose groups. An increase in this parameter would point to an acute irritating (inflammatory) response. However, such an irritant property could also be combined with a skin sensitizing potential of a test compound.

In conclusion, these results show that the test item Katalysator W AZ 5596-B has a sensitizing potential in mice after dermal application of a 2 % concentration.

Although it is not possible to calculate an exact EC value from the data obtained, it is clear that the EC value is in any case below 2%.

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

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

A sensitizing potential can be assumed from the increases in cell proliferation in the draining lymph nodes. On the basis of experiences by the author of the study using this method the "positive level" had been set to an increase in cell count index by 0.4 (i.e. index>1.4), which has been exceeded in the low and the mid dose group.

The "positive level" of ear swelling which is 2x 10E-2 mm increase, i.e. about 10% of the control values, has been exceeded in the mid and in the high dose group. An increase in this parameter does point to an acute irritant (inflammatory) response. Due to

the strong irritant property of the test item at these concentrations it is not quite clear if the cell proliferation is partly induced by this non-specific (inflammatory) reaction. On the other hand effects measured at the low concentration (2%) clearly point to a sensitizing property of the test item.

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

Migrated from Short description of key information:
A modified Local Lymph Node Assay (IMDS) indicate some sensitisation potential. At higher concentrations irritation was observed.

Justification for selection of skin sensitisation endpoint:
key study is 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

Due to the positive result of the LLNA a classification as R43 (GHS: Skin Sens 1a) ist justified