Pyridinium, 1-[4-[[4-[(4-amino-9,10-dihydro-9,10-dioxo-3-sulfo-1-anthracenyl)amino]cyclohexyl]amino]-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]-3-carboxy-, inner salt, sodium salt (1:3)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From December 09, 2015 to June 27, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

other company data

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The guinea pig maximization test (GPMT) is already available for the registration outside of EU. Therefore, LLNA is not conducted.

Test material

Test material information

Constituent 1

Reference substance name:

trisodium 1-[4-({4-[(4-amino-9,10-dioxo-3-sulfonato-9,10-dihydroanthracen-1-yl)amino]cyclohexyl}amino)-6-[(2,5-disulfonatophenyl)amino]-1,3,5-triazin-2-yl]-1λ⁵-pyridin-1-ylium-3-carboxylate

EC Number:

813-042-7

Cas Number:

1918149-24-2

Molecular formula:

C35H30N8O13S3 . 3Na

IUPAC Name:

trisodium 1-[4-({4-[(4-amino-9,10-dioxo-3-sulfonato-9,10-dihydroanthracen-1-yl)amino]cyclohexyl}amino)-6-[(2,5-disulfonatophenyl)amino]-1,3,5-triazin-2-yl]-1λ⁵-pyridin-1-ylium-3-carboxylate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male

Details on test animals and environmental conditions:

- Source: WEI XIN HANG
- Weight at study initiation: 352.1-379.6g
- Housing: Every five animals were housed in a stainless steel wire mesh cage
- Acclimation period: 7 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 50 ± 20%
- Photoperiod: 12-hrs dark / 12-hrs light

Study design: in vivo (non-LLNA)

Induction

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Concentration / amount:

0.1mL and 0.5mL of 10% CR SB08 for intradermal and epicutaneous, respectively

Day(s)/duration:

Day1 and Day7 for intradermal and epicutaneous, respectively

Adequacy of induction:

non-irritant substance, but skin pre-treated with 10% SDS

Challenge

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

0.5mL of 10% CR SB08

Day(s)/duration:

Day23

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Control group: five
Test group: ten

Positive control substance(s):

yes

Remarks:

α-Hexylcinnamaldehyde

Results and discussion

Positive control results:

The latest results showed that positive control animals had shown skin reactions of 1-2 and the sensitization rate of α-Hexylcinnamaldehyde (HCA) was 100%.

In vivo (non-LLNA)

Results

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.5 mL of 10% CR SB08

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

There was no emergence of erythema and edema on the skin of the observation period

Remarks on result:

no indication of skin sensitisation

Any other information on results incl. tables

Table 1. The body weight of the guinea pigs

Group	Sex	Animal ID.	Animal Weight (g)		Weight Change (g)
			Day 1	Day 26
Control group	Male	466	369.5	468.2	+98.7
		467	376.4	478.6	+102.2
		468	359.2	460.9	+101.7
		469	368.0	470.1	+102.1
		470	375.4	474.8	+99.4
Test group	Male	471	373.1	477.6	+104.5
		472	359.3	461.5	+102.2
		473	367.2	471.0	+103.8
		474	364.1	463.9	+99.8
		475	357.2	457.2	+100.0
		476	377.6	483.5	+105.9
		477	379.6	480.5	+100.9
		478	354.2	459.9	+105.7
		479	363.7	467.3	+103.6
		480	352.1	451.4	+99.3

Table 2. Individual skin reaction of guinea pigs

Group	Sex	Animal ID.	Grade of skin reaction^1		Percent of Animals Sensitized	Sensitizing Capacity^2
			24 ± 1 hrs	48 ± 1 hrs
Control group	Male	466	0	0	0%	Weak
		467	0	0
		468	0	0
		469	0	0
		470	0	0
Test group	Male	471	0	0	0%	Weak
		472	0	0
		473	0	0
		474	0	0
		475	0	0
		476	0	0
		477	0	0
		478	0	0
		479	0	0
		480	0	0

¹The skin reaction was grading according to “Magnusson and Kligman scale”.

²The sensitizing capacity of the test article was classified according to the “Scoring system of Kligman”.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 406 test method and “Scoring system of Kligman”, the sensitization rate of CR SB08 was 0% and CR SB08 caused weak sensitization in guinea pigs. Therefore, CR SB08 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the SuperLab Study Plan for M62-151100070001EN which is based on the SOP for the OECD 406 (SOPP-339) and OECD 406 (OECD, 1992). The results of this OECD 406 test for CR SB08 show that test reliability criteria was met.

A 10% CR SB08 was used for intradermal injection and occlusively patched, to ten test guinea pigs to induce sensitization. The normal saline was used for similar injection and occlusively patched to five control guinea pigs. Following a recovery period, animals received a challenge patch with 10% CR SB08. The skin sensitization reactions were scored at approximately 24 and 48 hours after patch removal. At the end of study, body weight increase was observed in all animals.During 48 hours after challenge exposure, 0% treated animals showed sensitization to the CR SB08. Under the conditions of this study,CR SB08 did not caused any sensitization in guinea pigs.