Pyridinium, 1-[2-[[4-[[3-[2-[6-amino-1-hydroxy-3-sulfo-5-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-2-naphthalenyl]diazenyl]-4-sulfophenyl]amino]-4-oxobutyl]sulfonyl]ethyl]-3-carboxy-, inner salt, sodium salt (1:4)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From January 4, 2019 to March 14, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-​Naphthalenesulfonic acid, 7-​amino-​4-​hydroxy-​, coupled with diazotized 2-​amino-​5-​[[2-​(sulfooxy)​ethyl]​sulfonyl]​benzenesulfonic acid and diazotized dehydrochlorinated 2-​amino-​4-​[[4-​[(2-​chloroethyl)​sulfonyl]​-​1-​oxobutyl]​amino]​benzenesulfonic acid, sodium salts

EC Number:

833-951-2

Cas Number:

2246977-27-3

Molecular formula:

C8H11NO9S3.C10H9NO4S.C12H17ClN2O6S2.Na

IUPAC Name:

2-​Naphthalenesulfonic acid, 7-​amino-​4-​hydroxy-​, coupled with diazotized 2-​amino-​5-​[[2-​(sulfooxy)​ethyl]​sulfonyl]​benzenesulfonic acid and diazotized dehydrochlorinated 2-​amino-​4-​[[4-​[(2-​chloroethyl)​sulfonyl]​-​1-​oxobutyl]​amino]​benzenesulfonic acid, sodium salts

Test material form:

solid: particulate/powder

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: WEI XIN HANG
- Weight at study initiation: 2.09 to 2.42 kg
- Housing: individually in a stainless steel wire mesh cage
- Water: ad libitum
- Acclimation period: 7 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 55 ± 15%
- Photoperiod: 12hrs dark / 12hrs light

Test system

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

Amount / concentration applied:

- Amount(s) applied (volume or weight with unit): 0.5 g test article was moistened with 0.5 mL normal saline

Duration of treatment / exposure:

4 hours

Observation period:

1, 24 ± 1, 48 ± 1, 72 ± 1 hrs

Number of animals:

3

Details on study design:

- Area of exposure: upper left and lower right dorsal Area
- Washing (if done): with distilled Water

After evaluating, test article had not been judged to be corrosive, so test article did not perform preliminary test and directly applied on main test.

Main study:
Before the test, 15 cm x 10 cm area over the dorsal region o f each test animal was shaved. The shoulder skin examination did not have any injury and marked the positions of the test and control area.

Gauze about 2.5cm x 2.5cm was evenly loaded with 0.5g test article and wetted by physiological saline. The gauzes were applied on the upper left and lower right dorsal skin of test animals.

Gauze patch about 2.5cm x 2.5cm was fully loaded with 0.5mL control article. The gauzes were applied on the upper right and lower left dorsal skin of test animals.

The gauzes loaded test article or control article were closely contact with dorsal skin of test animal through tightened bandage. After 4 hours working, the gauzes were taken off. The test area and control area were cleaned with water.

After gauzes taken off, the irritation response of rabbit skin was evaluated at 1, 24, 48, and 72 hours. Any skin irritation response, including erythema and edema was graded and recorded according to the ‘Grading system for skin reaction”.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 153, 154, 155

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal: 153, 154, 155

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Table 1. Body weight of the rabbits

Animal ID	Sex	Day 1	Day 4
153	male	2.37	2.42
154	male	2.29	2.30
155	male	2.09	2.19

Table 2. Individual skin reactions

Animal ID	Observation time(hours)		Skin grading (erythema/edema)          Test area   Control area Upper left   Lower right  Lower left  Upper right
153	1		0/0	0/0	0/0	0/0
153	24		0/0	0/0	0/0	0/0
153	48		0/0	0/0	0/0	0/0
153	72		0/0	0/0	0/0	0/0
154	1		0/0	0/0	0/0	0/0
154	24		0/0	0/0	0/0	0/0
154	48		0/0	0/0	0/0	0/0
154	72		0/0	0/0	0/0	0/0
155	1		0/0	0/0	0/0	0/0
155	24		0/0	0/0	0/0	0/0
155	48		0/0	0/0	0/0	0/0
155	72		0/0	0/0	0/0	0/0

Table 3. Primary irritation scores of rabbit skin irritation test

Animal ID	Irritation scores^a Test area Control area		Primary Iriitation Scores (PIS)^b
153	0	0	0
154	0	0	0
155	0	0	0
Primary irritatioin index (PII)^c = 0

^a Irritation cores were the sum of each rabbit irritation values at three time points and then divided by the total number of observation.

^b PIS = (scores of test area) – (scores of control area).

^c PII = Sum of PIS ÷ number of animals.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 404 test method and Evaluation table for single dermal irritation, the PII for CR SB33 was 0 and CR SB33 was categorized as non-irritant. Therefore, CR SB33 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the SuperLab Study Plan for MZ6 -181200022 which is based on the SOP for the OECD 404 (SOPP-305) and OECD 404 (OECD, 2015). The sites were grades for erythema and edema at 1, 24, 48 and 72 hours after the patches removal. No clinical signs of toxicity were found. Body weight changes were within the normal range in all animals. All animals was no formation of erythema, eschar or edema on the test area within 72 hours after patch removal. The Primary Irritation Index for CR SB33 was calculated to be 0. On the basis of the test results given above and according to Evaluation table for single dermal irritation, the response of CR SB33 was categorized as non-irritant.