Disodium [N-(2-chlorophenyl)-2-[(2-hydroxy-5-nitrophenyl)azo]-3-oxobutyramidato(2-)][3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]-7-nitronaphthalene-1-sulphonato(3-)]chromate(2-)

Endpoint summary

Description of key information

Acute administration by oral route LD50 > 5000 mg/kg body weight.

Acute administration by dermal route LD50 > 2000 mg/kg body weight.

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:

weight of evidence

Study period:

September 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Tif: RAIf

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ciba-Geigy premises
- Weight at study initiation: from 160 to 180 grams
- Fasting period before study: animals fasted overnight
- Housing: housed in group of 5 in Macrolon cages (type3)
- Diet (e.g. ad libitum): ad libitum NAFAG, Gossau SG
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1 °C
- Humidity (%): 55 ± 5 %
- Photoperiod (hrs dark / hrs light): 14 hours dark / 10 hours light

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 2% CMC

Doses:

3170, 4640, 6000, 7750 and 10000 mg/kg bw

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: physical condition

Statistics:

LD 50 including 95 % confidence limits were calculated by the probit analysis method (Goulden A., Methods of Statistical Analysis, John Wiley and Sons, 1960, 3rd printing, pages 404-408).

Sex:

male/female

Dose descriptor:

LD0

Effect level:

ca. 3 170 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 7 993 mg/kg bw

Based on:

test mat.

95% CL:

ca. 6 335 - ca. 10 085

Mortality:

Mortality observed for concentrations above 3170 mg/kg body weight.

Clinical signs:

other: Within 2 hours after treatment the rats in all dosage groups showed sedation, dyspnoea, exophthalmos, curved position, diarrhoea and ruffled fur.

Gross pathology:

No substance related gross organ changes were seen.

Rate of deaths

Dose mg/kg	Concentration % of formulation	N° of animals (M, male F, female)		Died within
				1 hour		24 hours		48 hours		7 days		14 days
		M	F	M	F	M	F	M	F	M	F	M	F
3170	30	5	5	0	0	0	0	0	0	0	0	0	0
4640	50	5	5	0	0	0	0	0	0	1	1	1	1
6000	50	5	5	0	0	0	0	0	2	0	2	0	2
7750	50	5	5	0	0	1	2	3	3	3	3	3	3
10000	50	5	5	0	0	0	4	1	5	1	5	1	5

 

Interpretation of results:

other: CLP criteria not met

Conclusions:

The acute oral LD50 of substance in rats of both sexes observed over a period of 14 days is 7993 (6335-10085) mg/kg bw.
LD50 = 5834.9 mg/kg bw based on active ingredient.

Executive summary:

Method

The test substance was tested for acute toxicity for oral route following a procedure similar to OECD 401.

The rats were kept at a room temperature of 22 ± 1 °C, at a relative humidity of 55 ± 5 % and on a 10 hours light cycle day. They received ad libitum rat food and water. Prior to treatment the animals were adapted for a minimum of 4 days and the initial body weight ranged from 160 to 180 grams.

During the treatment and observation period the animals were housed in groups of 5 in Macrolon cages (type 3).

Physical condition and rate of deaths were monitored throughout the whole observation period.

The substance was administered by oral intubation at the following concentrations: 3170, 4640, 6000, 7750 and 10000 mg/kg body weight. 5 rats per sex per dose were exposed at each concentration. Dyspnoea, exophtalmus, ruffled fur, curved body position and a transient diarrehea were seen.

Results

The LD50 observed in rats was 5834.9 mg/kg bw.

The animals were submitted to a necropsy whenever they died, survivors at the end of the observation period.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

7 993 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:

experimental study

Adequacy of study:

key study

Study period:

From September 28 to October 12, 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

not specified

GLP compliance:

no

Test type:

fixed dose procedure

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Perfection Breeders, Ace Animals, Nicholas Hel f
- Age at study initiation: 9 weeks
- Weight at study initiation: from 2.2 to 3.7 kg
- Housing: housed 1/cage in suspended wire mesh cages (30" x 18" x 18")
- Diet (e.g. ad libitum): fresh Purina rabbit chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 21°C

Type of coverage:

occlusive

Vehicle:

not specified

Details on dermal exposure:

TEST SITE
- Area of exposure: 200 square cm
- % coverage: ca. 10%

REMOVAL OF TEST SUBSTANCE
- Washing: warm tap water

TEST MATERIAL
- Amount(s) applied: rabbit #1 applied 6.7 cc; rabbit #2 applied 4.3 cc; rabbit #3 applied 4.1 cc; rabbit #4 applied 4.0 cc
- Constant volume or concentration used: no

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

2 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: dermal reactions were scored at 25 hours, 7 and 14 days. 14 days for signs of toxicity, pharmacological effects and mortality. Body weights were recorded pretest and in the survivors at 7 and 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight and gross pathology

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortalities observed.

Clinical signs:

other: One animal had lethargy, ptosis, emaciation, diarrhea, adipsia, anorexia and few feces. See table 1.

Gross pathology:

At necropsy, three animals were normal and one animal had white nodules on the liver

Other findings:

- Other observations: 24 Hour erythema scores could not be calculated due to the color of the material. All other dermal scores at all other times were zero.

Table 1 – Toxic Signs

ANIMAL # & SEX	HOUR	DAY
	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1-M													2	2	2
2-M
3-F				BQD	BD	BX	BX	BXQ	BXQ	BXQ	BDQ	BDQ	BDQW	BQW USX	BQW USX
4-F

CODE:

B = lethargy

D = diarrhea

Q = ptosis

S = adipsia

U = anorexia

W = emaciation

X = few feces

2 = white nasal discharge

Interpretation of results:

other: CLP criteria not met

Conclusions:

LD50 > 2000 g/kg bw.

Executive summary:

Method

The study was performed following a procedure similar to OECD 402.

New Zealand White rabbits, at least 8 weeks old, were equilibrated for at least one week. The animals were housed 1 per cage in suspended wire mesh cages (30" x 18" x 18"). The animal room was maintained at 20 - 21 °C and was kept clean in accordance with the standards of AAALAC.

24 hours prior to dosing, the fur was clipped from the backs of the animals. The clipped area was 200 square cm, approximately 10 %

of the body surface. Just prior to dosing, abrasions were made in one half of the rabbits. The abrasions, extending the length of the exposure site, scratched the stratum corneum but did not reach the derma or produce bleeding.

Two male and two female rabbits were dosed at 2000 mg/kg. The test material was applied once dermally to the prepared site under occlusive gauze patches. The test material was kept in contact with the skin for 24 hours, at which time the wrappings were removed. An estimate of the amount of material remaining was recorded. The exposure site was washed with warm tap water to remove excess material.

Dermal reactions were scored at 25 hours, 7 and 14 days by the Draize scoring system. The rabbits were observed daily for 14 days for signs of toxicity, pharmacological effects and mortality. Body weights were recorded pretest and in the survivors at 7 and 14 days.

Observations

All animals survived the test. At 14th day, the survivors were sacrificed. All animals were examined for gross pathology.

Results

LD50 > 2000 g/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

For the acute administration by oral route, the assessment of the substance is based on three studies.

The first study (Huntsman, 1977) was conducted according to a method similar to OECD 401. Ten rats per dose were administered with the following concentrations: 3170, 4640, 6000, 7750 and 10000 mg/kg body weight. The LD50 was 7993 mg/kg body weight.

The second study (Huntsman, 1984) was performed according to OECD 401 and a complete report is available. Ten rats were administered by gavage at a concentration of 5000 mg/kg body weight. At this concentration no mortality was observed.

The third study (Huntsman, 1973) was conducted following a method similar to OECD 401. Ten rats were administered at a concentration of 5000 mg/kg body weight. At the end of the observation period, no mortalities were observed.

Considering the results of all the studies the LD50 for the substance is considered to be > 5000 mg/kg body weight.

The acute dermal toxicity of the substance under registration was investigated in rabbits following a procedure similar to OECD 402. The substance was applied dermally for 24 hours under occlusive gauze patches at a concentration of 2000 mg/kg body weight. All animals survived during the 14 days post-treatment. One animal of the four used for the study, showed letargy, prosis, amaciation, diarrhea, adipsia, anorexia and few feces.

On the bases of the study result the LD50 can be considered > 2000 mg/kg.

Considering the overall results the substance has practically no acute toxicity for the two ways of exposure.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The oral LD50 value was established to be greater than 5000 mg/kg body weight, therefore the test substance is out of any classification limit for acute oral toxicity.

The dermal LD50 value was established to exceed 2000 mg/kg body weight, which exceeded the highest CLP classification limit.

In conclusion, the available experimental data are adequate for classification and labelling and the test substance is non classified for oral and dermal acute toxicity, according to the CLP Regulation (EC 1272/2008).