2-Naphthalenecarboxylic acid, 4-[2-[5-substituted-2-methoxyphenyl]diazenyl]-3-hydroxy-, calcium salt (2:1)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 June 2010 - 20 June 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material : J-37
- Substance type: red powder
- Physical state: powder
- Analytical purity: 99.3%
- Lot/batch No.: 91116
- Expiration date of the lot/batch: 16th April 2011
- Storage condition of test material: at room temperature in the dark in desiccators
- Stability under test conditions: stable

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approximately. 8 weeks old
- Weight at study initiation: Body weight variation did not exceed ± 20% of the sex mean.
- Fasting period before study: Overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages (MIV type; height 18 cm) containing sterlised sawdust as bedding material (Litalabo, S. P. P. S., Argenteuil, France) and paper as cage- enrichment (Enviro-dri, Wm. Lilico & Son (Wonham Mill Ltd), Surrey, UK).
- Diet : free access to pelleted rodent diet (SM R/M-Z from SSNIFF Spezialdiäten GmBH, Soest, Germany)
- Water : free access to tap water
- Acclimation period: 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual range: 19.7 - 21.5)
- Humidity (%): 40 - 70 (actual range: 38 - 75)
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artifical fluorescent light and 12 hours darkness per day

IN-LIFE DATES: 4 June 2010 - 20 June 2010

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 mL/ kg

Doses:

2000 mg/kg bw and 300 mg/kg bw

No. of animals per sex per dose:

6 animals at 300 mg/kg bw
3 animals at 2000 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Mortality/viability - twice daily. Body weights - Days 1 (pre-administration), 8 and 15 at death (if found deat or sacrificed after Day 1). Clinical signs - At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: yes. The moribund animal and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure.
- Other examinations performed: histopathology.

Clinical signs: The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1)

Statistics:

No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

other: LD50 cut-off value according to OECD 423 test guideline

Effect level:

500 mg/kg bw

Based on:

test mat.

Mortality:

All females at 2000 mg/kg were found dead or sacrificed in moribund condition on Day 2. No mortality occurred at 300 mg/kg (Table 1).

Clinical signs:

At the 2000 mg/kg dose level - Lethargy, flat/hunched posture, piloerection,watery discharge from the eye, pale appearance and/or hypothermia on Day 1 and/or 2 (all animals).
At 300 mg/kg dose level- Hunched posture and/or piloerection on Day 1 (all animals).

Body weight:

The body weight gain shown by the survivng animals over the study period was considered to be similar to the expected of normal untreated animals of the same age and strain (Table 2).

Gross pathology:

At the 2000 mg/kg dose level - watery-clear fluid in the abdominal cavity on one animal, a beginning stage of autolysis in two animals and hard contents of the stomach in all animals (Table 3).
At 300 mg/kg dose level- Pelvic dilation of the kidneys (one animal).

Any other information on results incl. tables

Table 1.Mortality data.

Test Day	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
Hours after treatment	0	2	4
Female 2000 mg/kg	-	-	-	3	-	-	-	-	-	-	-	-	-	-	-	-	-
Female 300 mg/kg	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Female 300 mg/kg	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Table 2. Body weight.

Sex/ dose level	Animal	Day 1	Day 2	Day 8	Day 15
Female 2000 mg/kg
	1	169	169	-	-
	2	169	171	-	-
	3	169	172	-	-
	Mean	169	171	-	-
	St.Dev	0	2	-	-
	N	3	3	0	0
Female 300 mg/kg
	4	159	-	194	209
	5	153	-	173	185
	6	160	-	188	195
	Mean	157	-	185	196
	St.Dev	4	-	11	12
	N	3	0	3	3
Female 300 mg/kg
	7	167	-	193	204
	8	150		175	186
	9	158	-	181	194
	Mean	158		181	194
	St.Dev	9	-	9	9
	N	3	0	3	3

 

   Table 3. Macroscopic findings.

Animal	Organ	Finding	Day of Death
Female 2000 mg/kg
1	General observations	Abdominal cavity: contains fluid, watery clear	Killed in extremis Day 2 after treatment.
	Stomach	Contents: hard
2	General observations	Beginning autolysis.	Spontaneous death Day 2 after treatment.
	Stomach	Contents: hard
3	General observations	Beginning autolysis.	Spontaneous death Day 2 after treatment.
	Stomach	Contents: hard
Female 300 mg/kg
4		No findings noted	Scheduled necropsy Day 15 after treatment.
5		No findings noted	Scheduled necropsy Day 15 after treatment.
6	Kidneys	Both sides; pelvic dilations.	Scheduled necropsy Day 15 after treatment.
Female 300 mg/kg
7		No findings noted	Scheduled necropsy Day 15 after treatment.
8		No findings noted	Scheduled necropsy Day 15 after treatment.
9		No findings noted	Scheduled necropsy Day 15 after treatment.

 

Applicant's summary and conclusion

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU