Hydrogen 9-amino-7-phenyl-5-(phenylamino)-4,10-disulphonatobenzo[a]phenazinium, disodium salt

Skin sensitisation

Administrative data

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: well performed GLP and OECD guideline stiudy

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen 9-amino-7-phenyl-5-(phenylamino)-4,10-disulphonatobenzo[a]phenazinium, disodium salt

EC Number:

276-817-3

EC Name:

Hydrogen 9-amino-7-phenyl-5-(phenylamino)-4,10-disulphonatobenzo[a]phenazinium, disodium salt

Cas Number:

72749-80-5

Molecular formula:

C28H20N4O6S2.2Na

IUPAC Name:

Benzo[a]phenazinium, 9-amino-7-phenyl-5-(phenylamino)-4,10-disulfo-, inner salt, sodium salt (1:2)

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands, B.V. Postbus 6174, NL - 5960 AD Horst / The Netherlands
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 19.2 - 22.7
- Housing: Single caging, Makrolon Type I, with wire mesh top, granulated soft wood bedding
- Diet (e.g. ad libitum): pelleted standard diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): artificial light 6.00 a.m. - 6.00 p.m.

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

0.0 (control), 12.5, 25, and 50%

No. of animals per dose:

4

Positive control substance(s):

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

Statistics:

Mean values and standard deviations were calculated for the body weights.

Results and discussion

Positive control results:

Test item concentration % (w/v) / S.I.

5 / 5.24
10 / 7.38
25 / 9.32

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

< 2.4

Test group / Remarks:

@ 50%

Remarks on result:

other: Test item concentration % (w/v) / S.I. 12.5% / 1.9 25.0% / 1.7 50.0% / 2.4

Results 2

Parameter:

other: disintegrations per minute (DPM)

Value:

<= 1 470.9

Test group / Remarks:

@ 50%

Remarks on result:

other: Test item concentration % (w/v) / DPM per lymph node [ 0.0% (control) / 603.3] [12.5% / 1135.7] [25.0% / 1035.9] [50.0% / 1470.9]

Any other information on results incl. tables

Test item concentration % (w/w)	Group	Measurement DPM	Calculation			Result
			DPM-BGa)	number of lymph nodes	DPM per lymph nodeb)	S.I.
---	BG I	43	---	---	---	---
---	BG II	38	---	---	---	---
---	1	4867	4827	8	603.3
12.5	2	9126	9086	8	1135.7	1.9
25	3	8328	8288	8	1035.9	1.7
50	4	11808	11768	8	1470.9	2.4

 

BG = Background (1 ml 5% trichloroacetic acid) in duplicate

1 = Control Group

2-4 = Test Group S.I. = Stimulation Index

a) = The mean value was taken from the figures BG I and BG II

b) = Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

The EC3 value could not be calculated, since all S.I.´s are below 3.

 

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the findings in this assay the test item was not a skin sensitiser.

Executive summary:

In the study the test item suspended in dimethylformamide was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 12.5, 25, and 50%.

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. Signs of local irritation such as ear redness could not be determined due to the intense colour of the test item.

In this study Stimulation Indices (S.I.) of 1.9, 1.7, and 2.4 were determined with the test item at concentrations of 12.5, 25, and 50% in dimethylformamide, respectively.