Hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 September 2016 to 11 October 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

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:

Hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

EC Number:

269-284-3

EC Name:

Hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

Cas Number:

68214-04-0

Molecular formula:

C44H24Cl2N14Na6O20S6

IUPAC Name:

hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

Test material form:

solid: particulate/powder

Details on test material:

Name Reactive Red 120
Appearance Red powder

In vivo test system

Test animals

Species:

mouse

Strain:

CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: AnLab Prague, Czech Republic
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 17.5-22.6 g
- Housing: individually in polypropylene cages suspended on stainless steel racks with Lignocel S3/4 bedding
- Diet: ssniff (ssniff Spezialdiäten GmbH, Germany) ad libitum
- Water: tap (drinking) water ad libitum
- Acclimation period: 6 days
- Indication of any skin lesions: none

ENVIRONMENTAL CONDITIONS
- Temperature (°C): of 22 ± 2°C
- Humidity (%): 50-60 %
- Air changes (per hr): not indicated
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

25, 50 and 100%

No. of animals per dose:

main study: 5 females/concentration

Details on study design:

PRE-SCREEN TESTS: on 2 females at 100%
- Compound solubility: NA (tested as such)
- Irritation: none
- Systemic toxicity: none
- Ear thickness measurements: 0.17-0.18 over 6 day period
- Erythema scores: no erythema observed

MAIN STUDY:
Day 1-3: application of 25µL of the appropriate dilution to the dorsum of each ear
Days 4 and 5: No treatment.
Day 6: 250µL of phosphate-buffered saline (PBS) containing 2 µCi (7.4 x 104 Bq) of 125I-iododeoxyuridine and 10-5M fluorodeoxyuridine was injected into all test and control mice via the tail vein.
Five hours later, the animals were sacrificed. The draining auricular lymph nodes from each ear were excised and pooled in PBS for each experimental group (pooled treatment group approach) and weighted.

- Body weight: on day 1 and 6
- Clinical signs: daily

Positive control substance(s):

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

Statistics:

EC3 value: by linear interpolation of points on dose-response curve, immediately above and below SI value of 3

Results and discussion

Positive control results:

positive controls were within historical control ranges

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

2.72

Test group / Remarks:

25%

Results 2

Key result

Parameter:

SI

Value:

3.56

Test group / Remarks:

50%

Results 3

Key result

Parameter:

SI

Value:

4.44

Test group / Remarks:

100%

Any other information on results incl. tables

Lymph node weight, DPM, SI, EC3 values.

	Lymph node	Number of
	weight (g)	lymph nodes	DPM	SI	EC3
Control	0.0394	10	589	-	33.25
Positive Control	0.0737	10	3961	6.21
Reactive Red 120 25%	0.0695	10	1603	2.72
Reactive Red 120 50%	0.0712	10	2099	3.56
Reactive Red 120 100%	0.0777	10	2618	4.44

 

Applicant's summary and conclusion

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

The substance is a skin sensitizer

Executive summary:

The substance was tested in a local lymphnode assay in 5 mice per dose (at 100, 50 and 25%). Following 3 days of treatment, on day 6 mice received 125I-iododeoxyuridine and 10-5M fluorodeoxyuridine and were terminated 5 hours later. The lymphnodes were excised, pooled and prepared cells suspensions were assessed for 125I incorporation. Simulation indices were calculated to be 2.72, 3.56 and 4.44 at 25, 50 and 100%. The EC3 was calculated to be 33.25%. Based on this finding the substance is considered a skin sensitizer (category 1B)