3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid

Endpoint summary

Description of key information

Acute toxicity: oral

The acute oral LD50 of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of 3,4,5,6-tetrachloro-2-(1,4,5,8- tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), when administered via oral route (gavage) in Sprague Dawley rats falls into the “Category not classified” as per criteria of CLP.

 

Acute toxicity: dermal

The acute dermal median lethal dose (LD50) of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, when administered to male and female Sprague Dawley rats was considered to be greater than 2000 mg/kg body weight. Thus according to CLP criteria for acute toxicity rating for the chemicals, it infers that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) does not classify as an acute dermal toxicant.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

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 experimental study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Principles of method if other than guideline:

The purpose of this study was to assess the Toxicological profile of test item to a single administration via oral route to Sprague Dawley rats. This study was designed to determine the acute toxicity at fixed dose levels by oral route of the test item.

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Test Item: 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No.18472-87-2)
- Source of test material: Sustainability Support Services (Europe) AB
- Lot/batch No.of test material: FG/15-16/0790
- Manufacturing Date: June, 2015
- Expiration date of the lot/batch: May, 2023
- Purity test date: No data
- Consistency: Solid, powder
- Colour: Red

RADIOLABELLING INFORMATION (Not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item was stored at ambient temperature.
- Stability under test conditions: No data
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was dissolved in distilled water. Test item was dissolved in distilled water.
The formulation was prepared fresh on the day of dosing.
- Preliminary purification step (if any): No data
- Final dilution of a dissolved solid, stock liquid or gel: No data
- Final preparation of a solid: No data

FORM AS APPLIED IN THE TEST (if different from that of starting material) : No data

OTHER SPECIFICS:
Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source National Institute of biosciences, Pune.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used.
- Weight at study initiation : Body weight range was 195.7 to 202.7 grams.
Body weights at the start :
Female
Mean : 200.21 g (= 100 %)
Minimum : 195.7 g (- 2.25 %)
Maximum : 202.7 g (+ 1.24 %)
Total No. of animals : 12
- Fasting period before study: Approximately 16 hours or more.
- Housing: The rats were housed in polycarbonate cages.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.6 to 23.2 degree centigrade.
- Humidity (%): 54.2% to 58.6%.
- Air changes (per hr): Ten to fifteen air changes per hour
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 21-09-2016 to 10-10-2016

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 300 mg/kg, 300 mg/kg , 2000 mg/kg and 2000 mg/kg
- Amount of vehicle (if gavage): No data
- Justification for choice of vehicle: No data
- Lot/batch no. (if required): No data
- Purity: No data

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.

DOSAGE PREPARATION (if unusual): No data

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No data

Doses:

Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg
Dose Group II : 2000 mg/kg

No. of animals per sex per dose:

Three females were used at each step.

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical Observations and General Appearance:
Animals were observed for clinical signs, mortality and morbidity, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.

Body weights:
Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.

Gross Pathology:
Necropsy was performed on found dead and surviving animals at the end of the study period on day 15. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique.

Histopathology:
Gross pathological examination revealed reddish discolouration of digestive tract from stomach to anal area with liquid faecal material. This staining (discolouration) of digestive tract and faecal matter was attributed to the red colour of the test item, hence no histopathology was considered.

Statistics:

No data

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

Group I
Step I : . All animals survived through the study period of 14 days.

Group I
Step II : Animals treated at the dose level of 300 mg/kg body weight survived through the study period of 14 days.

Group II
Step I : Animals treated at the dose level of 2000 mg/kg body weight : One animal died on day 1 after the dosing.

Group II
Step II : Animals treated at the dose level of 2000 mg/kg body weight : One animal died on day 1 after the dosing.

Clinical signs:

other: Group I Step I :Animals treated at the dose level of 300 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. Group I Step II :Animals treated at the dose level of 300 mg/kg body weight did not result in any

Gross pathology:

Gross pathological examination did not reveal any abnormalities in animals terminal sacrificed from 300 mg/kg and 2000 mg/kg dose groups.
Gross pathological examination revealed reddish discolouration of digestive tract from stomach to anal area with liquid faecal material in found dead animals from 2000 mg/kg dose group.

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

 

Test System : Sprague Dawley Rat

Sex : Female

Group I :   

 

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	300	No clinical signs observed	3	1,2,3	Day 0 to Day 14	0/3

 

 

Group I :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	300	No clinical signs observed	3	4,5,6	Day 0 to Day 14	0/3

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	2000	Diarrhoea (Reddish colour stools)	3	7 8,9	30 min. - 6 hrs. 1 hr. - 6 hrs.	1/3
		Reduced locomotor activity	3	7,9 8	4 hrs. - Day 1 4 hrs. - 6 hrs.
		Ataxic gait	3	7,9 8	4 hrs. - Day 1 4 hrs. - 6 hrs.

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	2000	Diarrhoea (Reddish colour stools)	3	10 11,12	30 min. - 6 hrs. 1 hr. - 6 hrs.	1/3
		Reduced locomotor activity	3	10,11 12	1 hr. - Day 1 2 hrs. - 6 hrs.
		Ataxic gait	3	10 11 12	2 hrs. - Day 1 4 hrs. - Day 1 2 hrs. - 6 hrs.

 

Staining of the stool is attributed to the red colour of the test item.

 

Table No. II

Mean Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Female

Group I :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	300	Mean	200.77	212.00	5.60	227.07	7.11	13.10
		± SD	2.03	2.92	1.14	2.51	0.53	0.66

 

Group I :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	300	Mean	198.43	210.80	6.23	227.93	8.13	14.87
		± SD	2.86	3.01	0.20	3.30	0.26	0.06

 

Group II :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	200.97	213.90	6.79	230.00	7.53	14.83
		± SD	1.40	2.12	0.46	1.84	0.21	0.27

 

Group II :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	2000	Mean	200.67	214.05	6.60	227.95	6.50	13.54
		± SD	1.91	2.33	0.26	1.48	1.85	2.26

 

 

Table No.III

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Female 

Group I :

 

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	300	1 - 3	TS	No abnormality detected

 Group I :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	300	4 - 6	TS	No abnormality detected

 

 Group II :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	7, 9	TS	No abnormality detected
		8	FD	Reddish discolouration of digestive tract from stomach to anal area with liquid faecal material

 

  Group II :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	2000	10, 11	TS	No abnormality detected
		12	FD	Reddish discolouration of digestive tract from stomach to anal area with liquid faecal material

 TS = Terminal sacrifice

 FD = Found dead

Interpretation of results:

other: Not Classified

Conclusions:

The acute oral LD50 of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of 3,4,5,6-tetrachloro-2-(1,4,5,8- tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), when administered via oral route (gavage) in Sprague Dawley rats falls into the “Category not classified” criteria of CLP.

Executive summary:

The study now reported was designed and conducted to determine the acute oral toxicity profile of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxo xanthen-9-yl)benzoic acid in Sprague Dawley rats. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I).

Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools), reduced locomotor activity and ataxic gait in all animals with onset at 30 minutes to 4 hours after the dosing. One animal died on day 1 after the dosing. As one mortality was observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools), reduced locomotor activity and ataxic gait in all animals with onset at 30 minutes to 2 hours after the dosing. One animal died on day 1 after the dosing. All animals from 300 mg/kg dose group survived through the study period of 14 days and two animals died from 2000 mg/kg dose group after the dosing. Staining of the stool is attributed to the red colour of the test item. Gross pathological examination did not reveal any abnormalities in animals terminal sacrificed from 300 mg/kg and 2000 mg/kg dose groups.Gross pathological examination revealed reddish discolouration of digestive tract from stomach to anal area with liquid faecal material in found dead animals from 2000 mg/kg dose group. The acute oral LD50 of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) was >2000 mg/kg body weight . Thus, it was concluded that the acute toxicity study of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), when administered via oral route in Sprague Dawley rats falls into the “Category not classified” criteria of CLP.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from experimental study report.

Acute toxicity: via dermal route

Link to relevant study records

Reference

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 experimental study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

This study was designed to determine the dermal LD50 of the test item (up to 2000 mg/kg) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: no
- Age at study initiation: Young adult male and female rats aged between 6 – 9 weeks were used.
- Weight at study initiation: The weight range of approximately 218.4 to 256.4 grams at initiation of dosing were used.
Body weights at the start :
Male
Mean : 246.68 g (= 100 %)
Minimum : 238.5 g (- 3.32 %)
Maximum : 256.4 g (+ 3.94 %)
Total No. of animals : 5
Female
Mean : 224.90 g (= 100 %)
Minimum : 218.4 g (- 2.89 %)
Maximum : 235.6 g (+ 4.76 %)
Total No. of animals : 5
- Fasting period before study : No data
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period : 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0 to 22.3 degree centigrade.
- Humidity (%): 55.7% to 59.6%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 28-09-2016 to 13-10-2016

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

(Distilled water)

Details on dermal exposure:

TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the total body surface area
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Constant volume or concentration used: yes
- For solids, paste formed: Yes

VEHICLE (not used)
- Amount(s) applied (volume or weight with unit):No data
- Concentration (if solution): No data
- Lot/batch no. (if required): No data
- Purity: No data

Duration of exposure:

24 hours

Doses:

A single dose of 2000 mg of the test item per kilogram of body weight was administered to ten rats (five males and five females).

No. of animals per sex per dose:

10 (5/sex).

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical Observations and General Appearance:
Animals were observed for clinical signs, mortality, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.
The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.

Evaluation of Dermal Reaction:
Dermal reaction was observed daily for study period of 14 days.

Body weights:
Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.

Gross Pathology:
Necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).

Histopathology:
No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

Statistics:

No data

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality observed

Mortality:

Sex : Male
Group I -
Animal treated at the dose level of 2000 mg/kg body weight: All animals survived through the study period of 14 days.

Sex : Female
Group I -
Animal treated at the dose level of 2000 mg/kg body weight: All animals survived through the study period of 14 days.

Clinical signs:

other: Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not resul

Gross pathology:

Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.

Other findings:

- Other observations:
Evaluation of Dermal Reaction
Sex : Male
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Sex : Female
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Male

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	5	1 - 5	0 - 14	0/5

 

Sex : Female 

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	5	6 - 10	0 - 14	0/5

 

 Table No. II

 Summary of Evaluation of Dermal Reaction

 

Test System : Sprague Dawley Rat

Sex : Male 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No dermal reaction observed	5	1 - 5	0 - 14	0/5

 

Sex : Female 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No dermal reaction observed	5	6 - 10	0 - 14	0/5

 

Table No.III 

Mean Body Weight and Percent Body Weight Gain (g) 

Test System : Sprague Dawley Rat

Sex : Male 

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	246.68	274.24	11.17	297.50	8.52	20.64
		± SD	7.30	8.65	1.27	6.88	1.91	2.54

 

 

Sex : Female

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	224.90	239.14	6.33	251.06	4.99	11.63
		± SD	6.59	7.46	0.72	7.69	1.36	0.81

 

 Table No.IV 

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat 

Sex : Male

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	1 - 5	TS	No abnormality detected

 

Sex : Female

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	6 - 10	TS	No abnormality detected

 TS = Terminal Sacrifice

 

Interpretation of results:

other: not classified

Conclusions:

The acute dermal median lethal dose (LD50) of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, when administered to male and female Sprague Dawley rats was considered to be greater than 2000 mg/kg body weight. Thus according to CLP criteria for acute toxicity rating for the chemicals, it infers that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) does not classify as an acute dermal toxicant.

Executive summary:

The study now reported was designed and conducted to determine the acute dermal toxicity profile of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3- oxoxanthen-9-yl)benzoic acid in Sprague Dawley rats.

The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days.   

Animals exhibited normal body weight gain through the study period of 14 days.

Gross pathological examination did not reveal any abnormalities attributable to the treatment.

It was concluded that the acute dermal median lethal dose (LD50) of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, when administered to male and female Sprague Dawley rats was found to be greater than 2000 mg/kg body weight. Thus according to CLP criteria for acute toxicity rating for the chemicals, it infers that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) does not classify as an acute dermal toxicant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from experimental study report.

Additional information

Acute toxicity: oral

In different studies, 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments in rodents, i.e. most commonly in rats for 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid. The studies are summarized as below:

 

The study now reported was designed and conducted to determine the acute oral toxicity profile of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxo xanthen-9-yl)benzoic acid in Sprague Dawley rats. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I).Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools), reduced locomotor activity and ataxic gait in all animals with onset at 30 minutes to 4 hours after the dosing. One animal died on day 1 after the dosing. As one mortality was observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools), reduced locomotor activity and ataxic gait in all animals with onset at 30 minutes to 2 hours after the dosing. One animal died on day 1 after the dosing. All animals from 300 mg/kg dose group survived through the study period of 14 days and two animals died from 2000 mg/kg dose group after the dosing. Staining of the stool is attributed to the red colour of the test item. Gross pathological examination did not reveal any abnormalities in animals terminal sacrificed from 300 mg/kg and 2000 mg/kg dose groups.Gross pathological examination revealed reddish discolouration of digestive tract from stomach to anal area with liquid faecal material in found dead animals from 2000 mg/kg dose group. The acute oral LD50 of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), when administered via oral route in Sprague Dawley rats falls into the “Category not classified” criteria of CLP.

 

Furthermore, the study reported was designed and conducted by Yu F. Sasaki et.al (Mutation Research 519 (2002) 103–119); to determine the acute oral toxicity profile of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxo xanthen-9-yl)benzoic acid (Phloxine B) in male DDY mice. No mortality was observed at 2000 mg/kg, hence the acute oral LD50 of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) was determined to be >2000 mg/kg body weight.

 

Moreover, the study was conducted to determine the acute oral toxicity profile of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxo xanthen-9-yl)benzoic acid in rats. (published in an RTECS database (2005)). 50% mortality was observed in treated rats at 8400.0 mg/kg bw. Therefore, the acute oral LD50 of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid was determined to be 8400.0 mg/kg body weight.

 

Thus, based on the above mentioned studies for target substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), the median lethal dose (LD50) value was found to be in the range of >2000.0 mg/kg b.wt. to 8400.0mg/kg b.wt. Thus, it was concluded that the acute toxicity study of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), when administered via oral route in rats and mice falls into the “Category Unclassified” as per criteria of CLP.

 

Acute toxicity: inhalation

According to column 2 of REACH Annex VIII, the acute toxicity inhalation study need not be conducted because exposure of humans via inhalation route is not likely taking into account the low vapour pressure of the substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, which is reported as 3.12E-021 Pa. Also considering the particle size distribution of the substance, the majority of the particles were found to be in the size of 75.0 micrometer which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid is highly unlikely. Therefore this study is considered for waiver.

 

Acute toxicity: dermal

In different studies, 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments in rodents, i.e. most commonly in rats for 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid along with the study available on structurally similar read across substance Disodium 2-(2,4,5,7-tetraiodo-6- oxido-3-oxoxanthen-9-yl)benzoate (CAS No. 16423-68-0). The studies are summarized as below:

 

The study now reported was designed and conducted at IIT, Pune (2016) to determinethe acute dermal toxicity profile of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3- oxoxanthen-9-yl)benzoic acid in Sprague Dawley rats.The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days.Animals exhibited normal body weight gain through the study period of 14 days.Gross pathological examination did not reveal any abnormalities attributable to the treatment.It was concluded that the acute dermal median lethal dose (LD50) of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, when administered to male and female Sprague Dawley rats was found to be greater than 2000 mg/kg body weight. Thus according to CLP criteria for acute toxicity rating for the chemicals, it infers that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) does not classify as an acute dermal toxicant.

Hence, based on the above mentioned studies for target substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2), the median lethal dose (LD50) value was found to be > 2000 mg/kg b.wt. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) does not classify as an acute dermal toxicant.

CLP Classification: “Unclassified”.

Justification for classification or non-classification

Based on the above mentioned studies for target substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) and to its read across substance, it can be found that LD50 oral and dermal value is greater than 2000 mg/kg b.wt. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the test substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) does not classify as an acute toxicant by the oral and dermal route.