Calcium bis[4-[[1-[[(2-methylphenyl)amino]carbonyl]-2-oxopropyl]azo]-3-nitrobenzenesulphonate]

Endpoint summary

Description of key information

Skin sensitization: Read-across, not sensitizing in LLNA, according OECD TG 429, GLP-compliant, 0.2, 2, 20 % (w/v) test substance, mouse, 2011, K1

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Further information are included as attachment in chapter 13 of the IUCLID dossier.

Reason / purpose for cross-reference:

read-across source

Parameter:

SI

Value:

1

Test group / Remarks:

negative control

Parameter:

SI

Value:

9.56

Test group / Remarks:

positive control

Parameter:

SI

Value:

1.18

Test group / Remarks:

0.2 % test group

Parameter:

SI

Value:

1.05

Test group / Remarks:

2 % test group

Parameter:

SI

Value:

1.58

Test group / Remarks:

20 % test group

Parameter:

other: disintegrations per minute (DPM)

Value:

84.46

Test group / Remarks:

negative control (mean)

Parameter:

other: disintegrations per minute (DPM)

Value:

807.23

Test group / Remarks:

positive control (mean)

Parameter:

other: disintegrations per minute (DPM)

Value:

99.78

Test group / Remarks:

0.2 % test group

Parameter:

other: disintegrations per minute (DPM)

Value:

88.33

Test group / Remarks:

2 % test group

Parameter:

other: disintegrations per minute (DPM)

Value:

133.59

Test group / Remarks:

20 % test group

Reference 2

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011

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)

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

Balb/c

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Breeding farm VELAZ S.LO., Kolec u Kladna, Czech Republic, RCH CZ 21760118
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 16.6 - 19.22 g
- Housing: 6/cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5d

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

Vehicle:

other: DAB 433 - mixture of40% dimethylacetamide, 30% acetone and 30% ethanol

Concentration:

0.2, 2, 20 % w/v

No. of animals per dose:

5/dose

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: 20% technically highest concentration
- Irritation: no effects or clinical symptoms
- Lymph node proliferation response:

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI >=3

TREATMENT PREPARATION AND ADMINISTRATION: 25 ul of the appropriate dilution to the dorsum of each ear once a day morning for 3 consecutive days. The application was performed very slowly by micropipette. Losses caused by draining from the ear must be minimized.

Day 1:
Open application of 25f.lL (in the morning, by pipette) of appropriate suspensions of the test substance, the vehicle alone or the positive control to the dorsum of each ear.

Days 2 and 3:
The application procedure repeated as carried out on day 1.

Days 4 and 5:
No treatment.

Day 6:
Injection 250 ul of phosphate-buffered saline (PBS) containing 7.425 x10^5 Bq 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: Dinitrochlorobenzene (DNCB)

Statistics:

For statistical calculations the software Statgraphic ® Centurion (version XV, USA) was used. Statistical evaluation of measured parameters was performed at first by applying the non-parametric Kruskal-Wallis test for the comparison of the measured effect in all treatment groups with the vehicle control group, as global test, and then the non-parametric two-group Mann-Whitney rank test (probability level 0.05) for all two-group comparisons.

Parameter:

SI

Value:

1

Variability:

negative control

Parameter:

SI

Value:

9.56

Test group / Remarks:

positive control

Parameter:

SI

Value:

1.18

Test group / Remarks:

0.2 % test group

Parameter:

SI

Value:

1.05

Test group / Remarks:

2 % test group

Parameter:

SI

Value:

1.58

Test group / Remarks:

20 % test group

Parameter:

other: disintegrations per minute (DPM)

Value:

84.46

Test group / Remarks:

negative control (mean)

Parameter:

other: disintegrations per minute (DPM)

Value:

807.23

Test group / Remarks:

positive control (mean)

Parameter:

other: disintegrations per minute (DPM)

Value:

99.78

Test group / Remarks:

0.2 % test group

Parameter:

other: disintegrations per minute (DPM)

Value:

88.33

Test group / Remarks:

2 % test group

Parameter:

other: disintegrations per minute (DPM)

Value:

133.59

Test group / Remarks:

20 % test group

Interpretation of results:

GHS criteria not met

Conclusions:

Under the given test conditions, the test substance does not elicit sensitising response in LLNA assay.

Executive summary:

The substance was tested for the assessment of skin sensitisation potential with the murine local lymph node assay with radionuclides in mice. Five mice per group were exposed on the dorsum of both ears once a day by test substance (0.2, 2, 20%) and control substances during 3 consecutive days. Primary proliferation of lymphocytes in the lymph node was evaluated and the SI determined. The animals exposed to the test substance at all concentrations showed no pathological skin reactions and no other negative clinical symptoms of intoxication. Comparison of Stimulation Indexes (SI) between all treated groups and control vehicle group revealed that the test substance did not cause significant increase in radioisotope incorporation into the DNA of dividing lymphocytes.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Skin sensitisation

As no reliable data on skin sensitisation was available for the test substance, a read-across to a study conducted with a structural anologue (CAS 12286-65-6) was performed. 

 

The substance was tested for the assessment of skin sensitisation potential with the murine local lymph node assay with radionuclides in mice. Five mice per group were exposed on the dorsum of both ears once a day by test substance (0.2, 2, 20%) and control substances during 3 consecutive days. Primary proliferation of lymphocytes in the lymph node was evaluated and the SI determined. The animals exposed to the test substance at all concentrations showed no pathological skin reactions and no other negative clinical symptoms of intoxication. Comparison of Stimulation Indexes (SI) between all treated groups and control vehicle group revealed that the test substance did not cause significant increase in radioisotope incorporation into the DNA of dividing lymphocytes.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The stimulation indices in the LLNA (OECD 429) did not show a dose dependent increase. As a result, the substance is not considered to be classified for skin sensitization under Regulation (EC) No. 1272/2008, as amended for the fifteenth time in Regulation (EC) No. 2020/1182.