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

Endpoint summary

Description of key information

Skin Irritation:

The dermal irritation study was designed and conducted to determine the dermal reaction of test chemical 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. The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) the end of 14 days observation period after patch removal. Hence, it was concluded that the substance is non-Irritating to the skin of rats under the experimental conditions tested.

Eye Irritation:

The Primary eye irritation potency was evaluated for the test material in three male rabbits. About 100 mg erythrosine was instilled into the lower conjunctival sac of the right eye. The left eyes of the animals served as controls. 5 minutes after application, the treated eye was washed with 300 ml distilled water and the cornea, iris and conjunctival mucosa of treated eyes were investigated at 12, 24, 48, and 72 hrs after treatment. The test chemical did not cause adverse ocular reactions at 12, 24, 48, and 72 hrs after treatment. Hence the test chemical was considered as not irritating to the eyes of treated rabbits.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

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:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

The study reported was designed and conducted to determine the dermal reaction of test chemical in Sprague Dawley rats.

GLP compliance:

yes

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- 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 217.7 to 254.5 grams at initiation of dosing were used.
Body weights at the start :
Male
Mean : 246.00 g (= 100 %)
Minimum : 235.8 g (- 4.15 %)
Maximum : 254.5 g (+ 3.46 %)
Total No. of animals : 5
Female
Mean : 223.30 g (= 100 %)
Minimum : 217.7 g (- 2.51 %)
Maximum : 227.9 g (+ 2.06 %)
Total No. of animals : 5
- 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): Room temperature was maintained at 20.0 to 22.3 degree centigrade.
- Humidity (%): Room humidity was maintained at 55.7% to 59.6%.
- Air changes (per hr): The animal room was independently provided with at least ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: From: To: No data

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): no

VEHICLE
- 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

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): No data
- Concentration (if solution): No data

POSITIVE CONTROL
- Amount(s) applied (volume or weight): No data
- Concentration (if solution): No data

Duration of treatment / exposure:

24 hrs.

Observation period:

14 days

Number of animals:

10 (5/sex).

Details on study design:

TEST SITE
- Area of exposure: 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.
- Time after start of exposure: 24 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : 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.

SCORING SYSTEM: Draize Method.

Irritation parameter:

erythema score

Basis:

mean

Time point:

14 d

Score:

0

Max. score:

10

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

14 d

Score:

0

Max. score:

10

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Overall result:
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.

Results based on erythema and edema score:
Irritation parameter : erythema score
Basis : animal:1-5
Time point : other: 0 - 14 d
Score : 0
Max. score : 0
Reversibility: no data
Remarks on result : no indication of irritation

Irritation parameter : edema score
Basis : animal:1-5
Time point : other: 0 - 14 d
Score : 0
Max. score : 0
Reversibility: no data
Remarks on result : no indication of irritation

Irritation parameter : erythema score
Basis : animal: 6-10
Time point : other: 0 - 14 d
Score : 0
Max. score : 0
Reversibility: no data
Remarks on result : no indication of irritation

Irritation parameter : edema score
Basis : animal:6-10
Time point : other: 0 - 14 d
Score : 0
Max. score : 0
Reversibility: no data
Remarks on result : no indication of irritation

Other effects:

Clinical Signs of Toxicity and Mortality
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. 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 did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days.

Body Weight
Sex : Male
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 10.00% and 17.02% respectively.

Sex : Female
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.14% and 11.41% respectively.

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

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

 

 

 

Individual Animal - Evaluation of Dermal Reaction

 

Test System : Sprague Dawley Rat

Sex : Male  

Group : I

Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
3	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
4	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
5	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

  

Sex : Female  

Group : I

Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
6	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
7	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
8	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
9	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
10	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

 

 

 

 

 

 

Interpretation of results:

other: Not irritating

Conclusions:

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) the end of 14 days observation period after patch removal. Hence, it was concluded that the substance is non-Irritating to the skin of rats under the experimental conditions tested .

Executive summary:

The dermal irritation study was designed and conducted to determine the dermal reaction of test chemical 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. The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) the end of 14 days observation period after patch removal. Hence, it was concluded that the substance is non-Irritating to the skin of rats under the experimental conditions tested.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from safety assessment report

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

Primary eye irritation potency was evaluated for the test material in three male rabbits.

GLP compliance:

not specified

Species:

rabbit

Strain:

not specified

Details on test animals or tissues and environmental conditions:

No data available

Vehicle:

not specified

Controls:

yes

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

5 minutes

Observation period (in vivo):

12, 24, 48, and 72 hrs

Number of animals or in vitro replicates:

3 male animals

Details on study design:

TEST SITE
- Area of exposure: lower conjunctival sac of the right eye
- % coverage: Not applicable
- Type of wrap if used: Not applicable

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Washed with 300 ml distilled water
- Time after start of exposure: 12, 24, 48, and 72 hrs after treatment

SCORING SYSTEM: cornea, iris and conjunctival mucosa parameters.

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 12, 24, 48, and 72 hrs

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No adverse ocular reactions was observed.

Interpretation of results:

other: not irritating

Conclusions:

The test chemical did not cause adverse ocular reactions at 12, 24, 48, and 72 hrs after treatment. Hence the test chemical was considered as not irritating to the eyes of treated rabbits

Executive summary:

Primary eye irritation potency was evaluated for the test material in three male rabbits. About 100 mg erythrosine was instilled into the lower conjunctival sac of the right eye. The left eyes of the animals served as controls. 5 minutes after application, the treated eye was washed with 300 ml distilled water and the cornea, iris and conjunctival mucosa of treated eyes were investigated at 12, 24, 48, and 72 hrs after treatment. The test chemical did not cause adverse ocular reactions at 12, 24, 48, and 72 hrs after treatment. Hence the test chemical was considered as not irritating to the eyes of treated rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation:

Various studies have been investigated for the test chemical to observe the potential for dermal irritation to a greater or lesser extent. The studies are based on in-vivo experiments conducted for target chemical in humans and rabbits that have been summarized as below;

 The dermal irritation study was designed and conducted to determine the dermal reaction of test chemical 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. The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) the end of 14 days observation period after patch removal. Hence, it was concluded that the substance is non-Irritating to the skin of rats under the experimental conditions tested.

 In next study, the test uses a single rabbit ear to indicate the Comedogenicity and irritancy of test chemical. The test substance was mixed in propylene glycol at a 9 to 1 dilution for testing unless otherwise indicated (10% concentration). A colony of New Zealand albino rabbits that have genetically good ears and free from mites were used. Three rabbits, weighing two to three kilograms, were used for each assay. Animals were housed singly in suspended cages and fed Purina Rabbit Chow and water ad libitum. Animals were maintained on a 12-hour light and 12-hour dark cycle. A dose of 1 ml of the test material was applied and spread once daily to the entire inner surface of once for five days per week for two weeks. The opposite untreated ear of each animal served as an untreated control. The irritancy produced by repeated application of the chemical on the surface epidermis in the rabbit ear is evaluated on a scale of 0 to 5. The grades are summarized as follows: 0 = No irritation; 1 = few scales, no Erythema; 2 = diffuse scaling, no Erythema; 3 = Generalized scaling with Erythema; 4 = Scaling, Erythema and Edema; 5 = Epidermal necrosis and slough. The chemical falls under Grade 0 (no irritation observed). Hence it can be concluded that the test chemical was not irritating to rabbit ears.

The above results were supported by another skin irritation study conducted on four male albino rabbits to study the irritancy parameter of the test material. 100 mg of test chemical was applied to two separate sites of abraded skin and another 1 inch square site served as control. All skin sites were covered by surgical Gauze and rubberized cloth for 24 hrs. After the treatment period, the covers were removed and test material was removed from the skin. Skin reactions were examined at the end of the 24 hr-exposure and 24 hrs thereafter and scored according to the modified method of Draize et al. (1944) as described by Lynch et al. (1978). The test chemical caused slight redness at the abraded and treated skin sites which was not present after 48 hrs. Hence the test chemical is considered as not a primary skin irritant to the skin of rabbits.

Based on the above summarized studies for target chemical, it can be concluded that the test chemical is unable to cause dermal irritation, and it can be concluded to be not irritating to skin. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Eye Irritation:

In different studies, the test chemical has been investigated for potential for ocular irritation to a greater or lesser extent. The studies are based on in- vitro and in-vivo experimental conducted in rabbits conducted for test chemical which have been summarized as below;

The Primary eye irritation potency was evaluated for the test material in three male rabbits. About 100 mg test chemical was instilled into the lower conjunctival sac of the right eye. The left eyes of the animals served as controls. 5 minutes after application, the treated eye was washed with 300 ml distilled water and the cornea, iris and conjunctival mucosa of treated eyes were investigated at 12, 24, 48, and 72 hrs after treatment. The test chemical did not cause adverse ocular reactions at 12, 24, 48, and 72 hrs after treatment. Hence the test chemical was considered as not irritating to the eyes of treated rabbits.

The above result was supported by another eye irritation study of test chemical conducted on six rabbits according to Draize et al. 0.2 ml of a 10% aqueous solution of test chemical was applied repeatedly (twice daily, 5d/week, for 4 weeks) to the conjunctival sac of one eye of a group of at least 6 albino rabbits. One hour after each application the eyes were examined for evidence of staining and irritation was scored according to Draize. Three days after the last application, two animals of each group were killed, upper lids were taken for microscopic examination. Eyeballs and posterior parts were examined grossly for evidence of staining or other abnormalities. The test chemical caused intense colouring of the iris and moderate conjunctival irritation. Staining lasted from 2 to 7 days. The test chemical did not cause severe eye irritation but resulted in spotty staining in some animals and very slight but uniform staining in other animals. Since the observed effects were not persisted, the test chemical was considered to be not irritating to the skin.

Further, an in-vitro ocular irritation potential of test article was determined according to the OECD 492. The MatTek EpiOcular™ model was used to assess the potential ocular irritation of the test articles by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to liquid test articles and controls for ~30 minutes, followed by a ~12 minute post-soak and approximately 2 hour recovery after the post-soak. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met, passing the acceptance criteria. The mean of OD for test chemical was determined to be 0.012.The mean % tissue viability of test chemical was determined to be 0.7%. Hence, under the experimental test conditions it was concluded that test chemical was considered to be irritating to the human eyes.

The in-vivo results were further supported by the primary eye irritation study conducted on male and female New Zealand Albino rabbits to evaluate the irritating nature of similar read across chemical. About 100mg/0.1g of the test compound was instilled into one eye each of three young adult rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7 and 10, 14 and 17 days after test item application. The instillation of the test item into the eye resulted in mild to moderate, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. A slight corneal opacity, affecting up to the whole area of the cornea, was also apparent in one animal from 24 hours to 10 days after treatment, a light to marked red staining was also observed in the treated eye of all animals during the first 7 days after treatment. All ocular effects were reversible and were no longer evident 17 days after treatment. Since the all the observed effects were fully reversible within the observation period, the test chemical was considered as not irritating to the eyes.

Even though the results from the in-vitro study indicate a possibility that the test chemical can be irritating to eye, but the results from the in vivo studies suggests that the test chemical is unable to cause ocular irritation. Considering the results of the in vivo studies, the test chemical can be considered as not irritating to eyes. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

Based on the available studies for target chemical, it can be concluded that the test chemical is unable to cause dermal irritation, and it can be concluded to be not irritating to skin. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Even though the results from the in-vitro study indicate a possibility that the test chemical can be irritating to eye, but the results from the in vivo studies suggests that the test chemical is unable to cause ocular irritation. Considering the results of the in vivo studies, the test chemical can be considered as not irritating to eyes. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.