Trisodium, bis [2-((E)-(4-hydroxy-2-oxido-3-((E)-(5-sulfonatonaphthalen-1-yl)diazenyl)phenyl)diazenyl)benzoate] chromate(3-)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From Aug. 1983 to Oct. 1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1984

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

1981

Deviations:

yes

Remarks:

Temperature fell below protocol standard. Humidity increased above protocol standard. Deviations did not affect purpose or integrity of study.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

400-320-4

EC Name:

-

Cas Number:

94933-05-8

Molecular formula:

not applicable

IUPAC Name:

chromium(3+) trisodium bis(2-(2-{4-hydroxy-2-oxido-3-[2-(5-sulfonatonaphthalen-1-yl)diazen-1-yl]phenyl}diazen-1-yl)benzoate)

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Cr1:CD(SD)BR strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston Road, Margate, Kent, England
- Females: nulliparous and non-pregnant
- Age at study initiation: 6 to 10 weeks
- Weight at study initiation: 155 to 294 g
- Fasting period before study: overnight
- Housing: in groups of 5 by gender, in solid floor polypropylene boxes with a bedding of softwood sawdust (Sawdust Marketing Company, Standon, Herts), replaced twice weekly
- Diet: ad libitum; SQC Rat and Mouse Maintenance Diet No.1, Expanded (Special Diets Services, Ltd., Stepfield, Witham, Essex, England)
- Water: ad libitum; mains water dispensed from glass water bottles (analytical test report demonstrates suitability)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18 to 23 °C
- Humidity: 42 to 80 %
- Air changes per hour: not specified
- Photoperiod (hrs dark / hrs light): 12/12

ALLOCATION:
- total randomisation procedure into two test groups

IDENTIFICATION:
- individually, using a system of indelible marking on the tail
- pink for 5000 mg/kg bw
- yellow for 4000 mg/kg bw

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 250 mg/ml; 400 mg/ml
- Amount of vehicle: 20 ml/kg bw; 10 ml/kg bw
- Justification for choice of vehicle: no information known to indicate that the stability of the test article would be adversely affected under the proposed conditions of administration

Doses:

Group 1 (animals 1 to 10): 5000 mg/kg bw
Group 2 (animals 11 to 20): 4000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females for each dose
Total: 20 test animals

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations: 1 and 4 hours after treatment then daily
- Frequency of weighing: days -1, 0, 7 and 14; at death
- Necropsy of survivors performed: yes
- Other examinations performed: overt signs of toxicity; behavioural changes

Statistics:

No statistical test indicated

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in the group administered 4000 mg/kg bw. 1 male animal died in the group administered 5000 mg/kg bw on the day following administration of the test item. No further mortality occurred until the end of the study period (14 days) when all animals were sacrificed.

Clinical signs:

other: All animals administered 4000 mg/kg bw demonstrated black staining around the anus on the day following administration. This symptom continued to the next day for 3 of these animals. On days 1 and 2 following administration, one animal demonstrated emacia

Gross pathology:

In the group administered 5000 mg/kg bw, subject 5M and 6F demonstrated dark red patches in all lobes of the lungs; subject 3M demonstrated dark red patches in all lobes of the lungs and light patches on the liver; and 7 animals demonstrated no abnormalities.
In the group administered 4000 mg/kg bw, no abnormalities were observed.

Any other information on results incl. tables

Table 1: Mortality and signs of reaction in test animals administered a single dose of the test item.

Group number	Dose level (mg/kg)	Signs of reaction	Number showing effects during the day of dosing (hour)		Number showing effects during the day of observation														Total mortality
			1	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	5000	normal	10	10	9	9	9	9	9	9	9	9	9	9	9	9	9	9	1/10
		death	0	0	1	0	0	0	0	0	0	0	0	0	0	0	0	0
2	4000	black staining around anus	0	0	10	3	0	0	0	0	0	0	0	0	0	0	0	0	0/10
		emaciation	0	0	1	1	0	0	0	0	0	0	0	0	0	0	0	0
		hunched posture	0	0	1	1	0	0	0	0	0	0	0	0	0	0	0	0
		piloerection	0	0	0	1	0	0	0	0	0	0	0	0	0	0	0	0
		normal	10	10	0	6	10	10	10	10	10	10	10	10	10	10	10	10
		death	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Table 2: Individual body weights of male and female test animals.

Group number	Gender	Dose level (mg/kg)	Animal number	Individual body weight on:
				Day -1 (pre-fast)	Day of dosing	Day 7	Day 14	Increment	At death
1	male	5000	1	230	220	*	*	*	182
			2	223	213	219	303	80
			3	214	205	220	301	87
			4	221	213	227	281	60
			5	217	209	221	289	72
2	male	4000	11	294	265	300	385	91
			12	257	238	260	337	80
			13	231	216	246	303	72
			14	281	285	321	402	121
			15	241	217	248	324	83
3	female	5000	6	170	161	173	189	19
			7	178	167	174	205	27
			8	177	170	183	213	36
			9	169	160	171	198	29
			10	155	149	159	180	25
4	female	4000	16	174	155	168	211	37
			17	186	168	175	231	45
			18	175	161	168	223	48
			19	182	164	179	217	35
			20	213	190	200	241	28

* animal died during study

Table 3: Individual findings at necropsy.

Group number	Dose level (mg/kg)	Animal number	Necropsy findings
1	5000	1M*, 2M, 4M, 7F, 8F, 9F, 10F	no abnormalities detected
		3M	lungs: dark red patches in all lobes
			liver: light patches
		5M, 6F	lungs: dark red patches in all lobes
2	4000	11M, 12M, 13M, 14M, 15M, 16F, 17F, 18F, 19F, 20F	no abnormalities detected

* animal died during study

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

The LD50 (oral, rat) of the test item was found to be greater than 5000 mg/kg bw.

Executive summary:

The acute oral toxicity of the substance was evaluated in an experimental study performed according to a procedure similar to the OECD Guideline 401 (1981). The test item was administered to male and female Sprague-Dawley rats of both sexes by oral gavage, at doses of 4000 and 5000 mg/kg bw (5 female and 5 male animals per dose). Animals were then monitored for mortality, clinical symptoms and body weight changes for 15 days. At the end of the study period, all remaining animals were sacrificed and subjected to gross pathological examination at necropsy.

No mortality was observed in the group administered 4000 mg/kg bw. 1 male animal died in the group administered 5000 mg/kg bw on the day following administration of the test item. No further mortality occurred until the end of the study period (14 days) when all animals were sacrificed. All animals administered 4000 mg/kg bw demonstrated black staining around the anus on the day following administration. This symptom continued to the next day for 3 of these animals. On days 1 and 2 following administration, one animal demonstrated emaciation, and one animal demonstrated hunched posture (a typical symptom of pain). One animal demonstrated piloerection 2 days after administration. From day 3 after administration, no symptoms were observed in this group until the end of the study period. No clinical signs were observed among the 9 surviving animals administered 5000 mg/kg bw. All surviving animals showed comparable body weight gain at the end of the study period. One male and one female from the group administered 5000 mg/kg bw test item demonstrated dark red patches in all lobes of the lungs; one male animal demonstrated dark red patches in all lobes of the lungs and light patches on the liver; and 7 animals demonstrated no abnormalities. In the group administered 4000 mg/kg bw test item, no abnormalities were observed. Based on these findings, the acute oral toxicity of the test item in rats of both sexes, observed over a period of 15 days, was estimated to be: greater than 5000 mg/kg bw. Therefore, it can be extrapolated that the LD50 (oral, rat) is greater than 5000 mg/kg bw.