Calcium 4-[(5-chloro-4-methyl-2-sulphonatophenyl)azo]-3-hydroxy-2-naphthoate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Calcium 4-[(5-chloro-4-methyl-2-sulphonatophenyl)azo]-3-hydroxy-2-naphthoate

EC Number:

230-303-5

EC Name:

Calcium 4-[(5-chloro-4-methyl-2-sulphonatophenyl)azo]-3-hydroxy-2-naphthoate

Cas Number:

7023-61-2

Molecular formula:

C18H13ClN2O6S.Ca

IUPAC Name:

calcium 4-[(5-chloro-4-methyl-2-sulfonatophenyl)diazenyl]-3-hydroxy-2-naphthoate

Test material form:

solid: nanoform

Details on test material:

- Aggregate state at room temperature: Solid
- Colour: Red
- Storage: At room temperature
- Expiration Date: April 04, 2010
- Manufacturing Date: March 21, 2003
- Identity: Graphtol-Rot P2B VP 2747
- Purity: 84.8% (of Pigment Red 48:2)
- Batch No.: Vers. EB 98-04 or Kron 393014
- Certificate of Analysis: AZ 212/e2

Test materials used in this dossier are all considered to fall under the definition of nano-materials according to the European Commission Recommendation 2011/696/EU as the synthesis and manufacturing of this pigment always yields particulate material with a fine particle size distribution.

Specific details on test material used for the study:

- Aggregate state at room temperature: Solid
- Colour: Red
- Storage: At room temperature
- Expiration Date: April 04, 2010
- Manufacturing Date: March 21, 2003
- Identity: Graphtol-Rot P2B VP 2747
- Purity: 84.8% (of Pigment Red 48:2)
- Batch No.: Vers. EB 98-04 or Kron 393014
- Certificate of Analysis: AZ 212/e2

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

The animals were kept conventionally. The experiment was conducted under standard laboratory conditions.
- Housing: single Cage Type: Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen)
- Bedding: granulated soft wood bedding (Harlan Winkelmann GmbH, D-33178 Borchen)
- Feed: pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
- Water: tap water, ad libitum, (Gemeindewerke, D-64380 Rossdorf)
- Environment: temperature 22 + 3°C
- Relative humidity 30 - 70%
- Artificial light 6.00 a.m. - 6.00 p.m.
- Age: 7 - 8 weeks (beginning of acclimatisation)
- Housing: The animals were distributed into the test groups at random and identified by cage number.
- Acclimisation: under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

6.25, 12.5, and 25 %. Concentrations were in terms of material as supplied.

No. of animals per dose:

4 females (nulliparous and non-pregnant)

Details on study design:

The test item was placed into a volumetric flask glass beaker on a tared balance and the vehicle (acetone: olive oil (4+1)) was quantitatively added. The test item concentrations were prepared individually. Homogeneity of the test item in the vehicle was maintained during treatment with the magnetic stirrer. The preparations were made freshly before each dosing occasion. The data of the non GLP study 1010111 performed with the same test item showed that the highest test item concentration, which could be technically used was a 25 % suspension. Even warming and sonicating could not achieve a higher concentration. With other vehicles, e.g. DMF, DMSO, ethanol: deionised water (7+3) or propylene glycol, higher concentrations could also not be achieved. Due to the colour of the test item no irritation effects were observable at the concentration of 25 %. The test item in the main study was, therefore, assayed at 6.25, 12.5, and 25 %. Concentrations were in terms of material as supplied.

Positive control substance(s):

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

Statistics:

not required

Results and discussion

Positive control results:

The validation- / positive control study was performed with alpha- Hexylcinnamaldehyde in acetone:olive oil (4+1) using CBA/CaOlaHsd mice in December 2006.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

1.05

Test group / Remarks:

25%

Results 2

Parameter:

SI

Value:

1.5

Test group / Remarks:

12.5%

Results 3

Parameter:

SI

Value:

1.37

Test group / Remarks:

6.25%

Results 4

Parameter:

other: disintegrations per minute (DPM)

Value:

7 700.01

Test group / Remarks:

25%

Results 5

Parameter:

other: disintegrations per minute (DPM)

Value:

10 840.9

Test group / Remarks:

12.5%

Results 6

Parameter:

other: disintegrations per minute (DPM)

Value:

9 978.42

Test group / Remarks:

6.25%

Results 7

Parameter:

other: disintegrations per minute (DPM)

Value:

7 315.07

Test group / Remarks:

Control group

Any other information on results incl. tables

All treated animals survived the scheduled study period and no systemic toxicity was observed. Due to the colour of the test item local irritation reactions such as ear redness could not be detected.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met