Merbromin

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Justification for type of information:

data is from experimental reports

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

To assess the toxicological profile of the test chemical on application as a single semi-occlusive dermal application to rats

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate

EC Number:

241-409-6

EC Name:

Disodium 2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate

Cas Number:

17372-87-1

Molecular formula:

C20H6Br4Na2O5

IUPAC Name:

disodium 2-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): Eosine
- Molecular formula (if other than submission substance): C20H8Br4O5.2Na
- Molecular weight (if other than submission substance): 691.858 g/mol
- Smiles notation (if other than submission substance): c1(c2c(oc3c1cc(Br)c(c3Br)[O-])c(c(=O)c(c2)Br)Br)c1c(cccc1)C(=O)[O-].[Na+].[Na+]
- InChl (if other than submission substance): 1S/C20H8Br4O5.2Na/c21-11-5-9-13(7-3-1-2-4-8(7)20(27)28)10-6-12(22)17(26)15(24)19(10)29-18(9)14(23)16(11)25;;/h1-6,25H,(H,27,28);;/q;2*+1/p-2
- Substance type: Organic
- Physical state: Solid

Test animals

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

- Source: Geniron Biolabs Pvt. Ltd., Bengaluru
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 11 to 12 Weeks
- Weight at study initiation: Females: 217.20 to 222.36 g
- Identification:By rat accession number. Identification of individual rats is by cage card and crystal violet colour body markings. The temporary body marking during acclimatization period was done with crystal violet. The rat accession numbers were allotted during the course of the study and was included in raw data and reported.
- Housing: Animals were housed individually in standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill. Additionally, polycarbonate rat huts were placed inside the cage as enrichment objects and were changed along with the cage once a week. Bedding: Steam sterilized corn cob was used and changed once a week along with the cage.
- Diet (e.g. ad libitum): Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, ad libitum
- Acclimation period: The animals were acclimatized six days for G1-DRF, eleven days for G2-DRF, thirteen days G3-DRF and fifteen days for G3-Main before treatment. Animals were observed once daily during acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 24°C
- Humidity (%): 56 to 67 %
- Air changes (per hr): air conditioned with adequate fresh air supply (12.9 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 22 June 2018 To: 31 August 2018

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: Milli-Q water

Controls:

not specified

Amount / concentration applied:

DRF = 200, 100, 2000 mg/kgbw
Main = 2000 mg/kg

Duration of treatment / exposure:

24 hourd

Observation period:

treatment site was observed 24, 48 and 72 hours after removal of test chemical using the Draize criteria

Number of animals:

G1 – DRF - 1
G2 – DRF - 1
G3 – DRF and G3-Main – 3

Details on study design:

TEST SITE
- Area of exposure: clipped skin of the torso
- % coverage: 10% of the body surface of the animal (semi-occlusive).
- Type of wrap if used: The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso
REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: After the 24 hours contact period, the dressing was removed and the applied area was washed with deionized water and wiped dry using clean towels
OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : All the rats were observed for clinical signs of toxicity and mortality
for 14 days post application. In addition, the treatment site was observed at 24, 48 and 72 hours after removal of test item using the Draize criteria
SCORING SYSTEM:
- Method of calculation: Draize method

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

There were no skin reactions at the site of application at 24, 48 and 72 hours after test patch removal (as per Draize method).

Other effects:

There were no clinical signs and pre-terminal deaths (mortality) observed during the study

Any other information on results incl. tables

Dose Range Finding Study -DRF

Group & Dose (mg/kg body weight)

Date and time of application

Rat Number

Sex

Initial Bwt (g)

Quantity (mg) applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G1 and 200

28 JUNE 2018 10:23 to 10: 24 AM

Rw223

F

238.66

221.66

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Group & Dose (mg/kg body weight)

Animal Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G1 and 200

Rw223

F

N

N

N

N

N

N

N

N

N

N

NAD

Group & Dose (mg/kg body weight)

Date and time of application

Rat Number

Sex

Initial Bwt (g)

Quantity (mg) applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G2 and 1000

03 July 2018 and 11.24 AM

Rw224

F

217.20

217

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Group & Dose (mg/kg body weight)

Animal Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G2 and 1000

Rw224

F

N

N

N

N

N

N

N

N

N

N

NAD

Group & Dose (mg/kg body weight)

Date and time of application

Rat Number

Sex

Initial Bwt (g)

Quantity (mg) applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G3 and 2000

05 July 2018 and 11.30 AM

Rw225

F

221.40

442

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Group & Dose (mg/kg body weight)

Animal Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G3 and 2000

Rw225

F

N

N

N

N

N

N

N

N

N

N

NAD

Main study

Group & Dose (mg/kg body weight)

Date and time of application

Rat Number

Sex

Initial Bwt (g)

Quantity (mg) applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G3 and 2000

07 July 2018 and 11:32 AM and 11:33 AM

Rw226

F

219.14

438

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Rw227

F

222.36

444

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Group & Dose (mg/kg body weight)

Animal Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G3 and 2000

Rw226

F

N

N

N

N

N

N

N

N

N

N

NAD

Rw227

F

N

N

N

N

N

N

N

N

N

N

NAD

N: Normal

h: hour

min: minutes

NAD: No abnormality detected

Er: Erythema

Ed: Edema Score

0: No Erythema / Edema

*: Clinical signs; @: Skin scoring as per Draize method (approximately 24, 48 and 72 hours) after test patch removal

Applicant's summary and conclusion

Interpretation of results:

other: not irritating

Conclusions:

There were no skin reactions at the site of application at 24, 48 and 72 hours after test patch removal (as per Draize method). The mean erythema and edema scores were 0.0 at all observation times. Hence, it was concluded that the test chemical was "Not Irritating" to the skin of Wistar rats under the experimental conditions tested and classified as “Category-Not Classified” as per CLP Classification.

Executive summary:

A study designed and conducted to determine the dermal reaction profile of the test chemical in Wistar rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures.Wistar rats were tested with 200 mg/kg, 1000 mg/kg and 2000 mg/kg with 1 female for the dose range finding study, followed by additional 2 females for main study at the dose of 2000 mg/kg body weight. Based on the individual body weight, the test item at the dose of 200 mg/kg, 1000 mg/kg and 2000 mg/kg body weight was weighed on an aluminium foil and made into a paste by adding sufficient volume of the Milli-Q water and completely transferred on to the cotton gauze (size: Females: 8 x 5 cm of 6 ply) and applied directly (semi-occlusive) to the clipped skin of the rat to cover about 10% of body surface of the rat. It was secured in position by adhesive tape wound around the torso. The

test item contact period with the skin was for 24 hours. After the 24 hours contact period, the dressing was removed and the applied area was washed with deionized water and wiped dry using clean towels.

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed at 24, 48 and 72 hours after removal of test item using the Draize criteria.

There were no clinical signs of toxicity and mortality. There were no skin reactions at the site of application at 24, 48 and 72 hours after test patch removal (as per Draize method). The mean erythema and edema scores were 0.0 at all observation times. Hence, it was concluded that the test chemical was "Not Irritating" to the skin of Wistar rats under the experimental conditions tested and classified as “Category-Not Classified” as per CLP Classification.