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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from study report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

The objective of this acute dermal toxicity study was to assess the toxicological profile of the test item on application as a single semi-occlusive dermal application to rats.

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

no

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:

- IUPAC Name: Disodium 2-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate
- InChI: 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
- Smiles: c1(c2c(oc3c1cc(Br)c(c3Br)[O-])c(c(=O)c(c2)Br)Br)c1c(cccc1)C(=O)[O-].[Na+].[Na+]
- Molecular formula :C20H6Br4Na2O5
- Molecular weight :691.858 g/mol
- Substance type:Organic
- Physical state:Brownish powder
- Purity as per Certificate of Analysis:95.022 %
- Lot No.:L168831601
- Manufactured date:JAN-2016
- Retest date:DEC-2020
- pH:6.55 at 26 °C
- Density:Pour density :0.781 g/cm3 @ 27.05 °C; Tap density :0.97 g/cm3 @ 27.05 °C
- Storage conditions:Ambient (+15 to +25 °C )

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- 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

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

other: Milli-Q water

Details on dermal exposure:

TEST SITE
- Area of exposure: the hair on the dorsolateral thoracic surface of the skin was clipped (approximately 10 x 8 cm) with an electric clipper.
- % coverage: about 10% of body surface of the rat.
- Type of wrap if used: The cotton gauze was secured in position by adhesive tape wound around the torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the applied area was washed with deionized water and wiped dry using clean towel.
- Time after start of exposure:24 hours

TEST MATERIAL
- For solids, paste formed: yes, the test item at dose of 200 (G1-DRF), 1000 (G2-DRF) and 2000 (G3-DRF and Main) mg/kg body weight, was weighed on an aluminium foil and made into a paste by adding sufficient volume (about 0.1 mL, 0.2 mL, 0.3 mL and 0.4 mL) of Milli-Q water.

Duration of exposure:

24 hours

Doses:

Three treatment group
G1 – DRF
G2 – DRF
G3 – DRF and G3-Main

No. of animals per sex per dose:

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

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical examination and pre-terminal deaths - The animals were observed for clinical signs and pre-terminal deaths (mortality) once during first 30 minutes after application, and at hourly intervals for 6 hours after application on the day of treatment (day 1) and once daily during Days 2 to 15. In addition, treatment site at was observed 24, 48 and 72 hours after removal of test item using the Draize criteria (Refer Annexure 4 of this report). All rats were observed for changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

- Body weights - Individual body weights of animals were recorded on test days 1 (Pre-application), 8 (7 days post application), and 15 (14 days post application).

- Necropsy of survivors performed: yes, at the end of the observation period, all rats were euthanised and exsanguinated under isoflurane anesthesia and subjected to detailed necropsy by an experienced prosector and the findings were recorded. Microscopic examination was not carried out as no gross pathological changes were observed.

Statistics:

not specified

Results and discussion

Preliminary study:

Dose range finding study - As there was no acute dermal toxicity data available, an initial dose of 200 mg/kg body weight was tested in 1 female rat (dose range finding study) followed by 1000 mg/kg and 2000 mg/kg (dose range finding study). Based on the outcome of these dose range-finding studies, the main study was conducted with 2 further animals to confirm the classification. There was no test item-related mortality. The subsequent dosing was done approximately 2-4 days after the previous dosing.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no pre-terminal deaths (mortality) observed during the study.

Clinical signs:

other: There were no clinical signs observed during the study.

Gross pathology:

No abnormality was detected at necropsy.

Other findings:

not specified

Any other information on results incl. tables

TABLE 1.	Individual body weight, body weight changes and pre-terminal deaths
Group and				S				Body weight (g)				Pre-terminal
Dose		Rat										deaths
						Initial
				e			8th	Weight change		15th	Weight change
(mg/kg body weight)		No.		x		(Day 1 - at	day	(day 8 – Initial)		day	(day 15 – Initial)
						treatment)
G1		Rw223
200				F		221.66	223.87	2.21		225.29	3.63	0
DRF
G2		Rw224
1000				F		217.20	220.14	2.94		222.61	5.41	0
DRF
G3		Rw225
2000				F		221.00	223.12	2.12		226.92	5.92	0
DRF
G3		Rw226		F		219.14	220.89	1.75		224.83	5.69	0
2000
Main study		Rw227		F		222.36	225.16	2.8		230.45	8.09	0
DRF: Dose Range		Finding	F: Female

 

 

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

Based on the present study results, the acute dermal LD50 of the given test chemical is >2000 mg/kg body weight in female Wistar rats. The test item does not classified for acute dermal toxicity.
CLP classification "Not classified”.

Executive summary:

The acute dermal toxicity study was conducted as per OECD Guideline 402 (Acute Dermal Toxicity) for the given test chemical 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 in Wistar rats.

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. There were no clinical signs of toxicity and mortality. There was no skin reaction observed at test item applied area. Body weight was measured on days 1, 8 and 15 and all rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy. There were no abnormalities detected at the necropsy.

Based on the present study results, the acute dermal LD50 of the given test chemical is >2000 mg/kg body weight in female Wistar rats. The test item does not classified for acute dermal toxicity. CLP classification "Not classified”.