Disodium hydrogen bis[3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]naphthalene-1-sulphonato(3-)]chromate(3-)

Endpoint summary

Description of key information

Acute oral toxicity, rats (m/f), single oral exposure (OECD 401), LD50 > 2000 mg/kg bw
Acute dermal toxicity, analogue substance, rats(m/f), semiocclusive,  LD50 > 2000 mg/kg bw

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study following official guidelines, GLP compliant, performed on a similar substance

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Age at study initiation: 8 weeks for male, 9-10 weeks for female- Weight at study initiation: average 199g for male and average 173g for female- Acclimation period: 5 days adaptationThe female were nulliparous and non-pregnant.5 animals per sex were used

Doses:

single oral administration equal to 2000 mg/kg

No. of animals per sex per dose:

5 animals per sex

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed

Clinical signs:

other: After single-dosing of 2000 mg / kg no poisoning phenomena were observed during the 14-day observation time. Eight hours up to a day after the administration-of the tested substance a corresponding blue color of the dye administered was found in the faece

Gross pathology:

pathologically unremarkable

Interpretation of results:

practically nontoxic

Remarks:

Migrated information

Conclusions:

The substance was tested following OECD 401 for acute oral toxicity showing a LD50 > 2000 mg/kg.

Executive summary:

The substance was tested for acute toxicological studies in male and female wistar rats with single oral administration.

The LD50 (LD0) was found to be for male and female rat greater than 2000 mg/ kg. The administered dose resulted in blue dyed faeces. Poisoning phenomena were not observed. Body weight was not influenced in male and female rats. No mortality was observed. All animals were pathologically unremarkable.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2012-06-12 to 2012-08-21

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study performed on analogue substance

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Full barrier in an air-conditioned room- Temperature: 22 +/- 3 °C- Relative humidity: 55 +/- 10%- Artificial light, sequence being 12 hours light, 12 hours dark- Air change: 10 x / hour- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. . 0715)- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)- The animals were kept individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 261111)- Certificates of food, water and bedding are filed at BSL BIOSERVICE- Adequate acclimatisation period (at least five days) under laboratory conditions

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

Aqua ad injectionem (Diprom, lot no. 10952-1, expiry date: 09/2013)

Details on dermal exposure:

Preparation of the Animals:The animals were marked for individual identification by tail painting.Approximately 24 hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper. Care was taken to avoid abrading the skin, and only animals with healthy intact skin were used.No less than 10% of the body surface was cleared for the application.Prior to the application a detailed clinical observation was made of all animals.Application:The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface.The test item was held in contact with the skin by a dressing throughout a 24-hour period. The dressing consisted of a gauze-dressing and non-irritating tape and was fixed with an additional dressing in a suitable manner.

Duration of exposure:

The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was removed using aqua ad injectionem.

Doses:

The test item was applied at a single dose of 2000 mg/kg body weight to each animal considering the active component content of 86.5%.

No. of animals per sex per dose:

5 male and 5 female

Control animals:

not required

Details on study design:

Observation period:All animals were observed for 14 days after dosing.Weight Assessment:The animals were weighed on day 1 (prior to the application) and on days 8 and 15.Clinical Examination:careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.Pathology:At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial) at the dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded and in case of findings the tissues were preserved for a possible histopathological evaluation. The preserved tissues of which no histopathological evaluation was made will be discarded 3 months after the release of the final report unless otherwise agreed upon with the sponsor.Evaluation of Results:Individual reactions of each animal were recorded at each time of observation.Toxic response data were recorded by sex and dose level.Nature, severity and duration of clinical observations were described.The body weight changes were summarised in a tabular form.Necropsy findings were described.

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Sex:

male/female

Dose descriptor:

approximate LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality was observed

Clinical signs:

other: No treatment-related effects were observed.

Gross pathology:

No treatment-related effects were observed.

Other findings:

No erythema or oedema was observed.

Table Clinical Signs of Systemic Toxicity – Individual Data - Males:

Animal No. / Sex / Dose	Time of Observation (Post-Dose)	Observations (for Signs of Dermal Irritation, see Table 5)
21/ male / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
22/ male / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
23/ male / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
24/ male / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
25/ male / 2000 mg/kg bw	during the whole observation period	no signs of toxicity

Table Clinical Signs of Systemic Toxicity – Individual Data – Females:

Animal No. / Sex / Dose	Time of Observation (Post-Dose)	Observations (for Signs of Dermal Irritation, see Table 6)
26/ female / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
27/ female / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
28/ female / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
29/ female / 2000 mg/kg bw	during the whole observation period	no signs of toxicity
30/ female / 2000 mg/kg bw	during the whole observation period	no signs of toxicity

Table Skin Irritation – Individual Data – Males:

Day after Start of Application	Animal No. 21		Animal No. 22		Animal No. 23		Animal No. 24		Animal No. 25
	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments
day 2	0/0	R	0/0	R	0/0	R	0/0	R	0/0	R
day 3	0/0	R	0/0	R	0/0	R	0/0	R	0/0	R
day 4	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 5	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 6	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 7	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 8	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 9	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 10	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 11	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 12	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 13	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 14	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 15	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
Comments: E = erythema; O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404; R = residues of test item

Table Skin Irritation – Individual Data – Females:

Day after Start of Application	Animal No. 26		Animal No. 27		Animal No. 28		Animal No. 29		Animal No. 30
	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments
day 2	0/0	R	0/0	R	0/0	R	0/0	R	0/0	R
day 3	0/0	R	0/0	R	0/0	R	0/0	R	0/0	R
day 4	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 5	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 6	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 7	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 8	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 9	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 10	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 11	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 12	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 13	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 14	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
day 15	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf	0/0	nsf
Comments: E = erythema; O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404; R = residues of test item

Table Absolute Body Weights in g and Body Weight Gain in %:

Dose: 2000 mg/kg body weight
Animal No. / Sex	g Day 1	g Day 8	g Day 15	% Day 1-15
21 / male	250	271	306	22
22 / male	239	257	288	21
23 / male	228	243	270	18
24 / male	240	255	282	18
25 / male	254	277	303	19
26 / female	215	222	228	6
27 / female	208	216	229	10
28 / female	214	226	235	10
29 / female	218	242	250	15
30 / female	221	226	230	4

Table Macroscopic Findings - Individual Data – Males and Females:

Dose: 2000 mg/kg bw
Animal No. / Sex	Organ	Macroscopic Findings
21 / male	-	nsf
22 / male	-	nsf
23 / male	-	nsf
24 / male	-	nsf
25 / male	-	nsf
26 / female	-	nsf
27 / female	-	nsf
28 / female	-	nsf
29 / female	-	nsf
30 / female	-	nsf

nsf = no specific findings

Table LD50:

Dose (Unit)	Number of Animals Investigated	Number of Intercurrent Deaths	LD50
2000 mg/kg bw	5 males	0	> 2000 mg/kg bw
2000mg/kg bw	5 females	0	> 2000 mg/kg bw

bw = body weight

Interpretation of results:

practically nontoxic

Remarks:

Migrated informationCriteria used for interpretation of results: OECD GHS

Conclusions:

Under the conditions of the present study, single dermal application of the analogue substance to rats at a dose of 2000 mg/kg body weight was associated with no mortality and neither signs of toxicity nor signs of irritation.The dermal LD50 was determined to be > 2000 mg / kg body weight.

Executive summary:

LD₅₀: > 2000 mg /kg bw

Species/strain: WISTAR Crl: WI(Han) rats

Vehicle (moistening): aqua ad injectionem

Number of animals: 5 male and 5 female

Duration of exposure: 24 hours

Method: OECD 402, EC 440/2008, Method B.3, OPPTS 870.1200

Results per Step

Sex	Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths
male	2000	5	0
female	2000	5	0

Signs of toxicity related to dose level used, time of onset and duration: No treatment-related effects were observed.

Effect on organs (related to dose level): No treatment-related effects were observed.

Signs of irritation: No erythema or oedema was observed.

Conclusion

Under the conditions of the present study, single dermal application of the test analogue substance to rats

at a dose of 2000 mg/kg body weight was associated with no mortality and neither signs of toxicity nor signs of irritation.

The dermal LD50 was determined to be > 2000 mg / kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

There are valid in vivo data available for the assessment of the acute toxicity of the test substance.

Oral exposure pattern is covered by one study on the substance with a different degree of neutralisation, dermal toxicity is assessed through a very similar substance (analogue substance) and inhalation is not considered as a potential exposure pattern based on the vapour pressure. The oral toxicity has been performed as limit test at 2000 mg/Kg bw as well as the dermal toxicity. No mortality was observed in both studies nor systemic toxicity effects (no pathological findings or clinical signs rather than blue dyed faeces).

Justification for classification or non-classification

According to the CLP Regulation (EC n. 1272/2008), table 3.1.1, Acute toxicity hazard categories and acute toxicity estimates (ATE) defining the respective categories:

For Acute toxicity oral route:

Category 1: ATE <= 5 mg/kg bw

Category 2: 5 < ATE <= 50 mg/kg bw

Category 3: 50 < ATE <= 300 mg/kg bw

Category 4: 300 < ATE <= 2000 mg/kg bw

For the dermal route:

Category 1: ATE <= 50 mg/kg bw

Category 2: 50 < ATE <= 200 mg/kg bw

Category 3: 200 < ATE <= 1000 mg/kg bw

Category 4: 1000 < ATE <= 2000 mg/kg bw

The LD50 of the test substance was determined to be greater than 2000 mg/kg bw for noth oral and dermal exposure. Therefore, the test substance is not classified for Acute toxicity by oral and dermal exposure.