Ammonium 6H-dibenzo[c,e][1,2]oxaphosphinin-6-olate 6-oxide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 July 2011 to 3 October 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

DOPO-OX-Ammonium

IUPAC Name:

DOPO-OX-Ammonium

Details on test material:

Name of test material (as cited in study report): "DOPO-OX-Ammonium"
Chemical name: 9,10-dihydro-10-hydroxy-9-oxa-10-phosphaphenanthren-10-on- resp. 10-oxid-ammonium salt, corrected later to: 9,10-dihydro-10-hydroxy-9-oxa-10-phosphaphenanthren-10-oxid-ammonium salt
Trade name: DOPO-OX-Ammonium
Hill formula: C12H12NO3P
Lot No.: DAS-1101
Appearance: White to slight yellow solid
Conditions of storage: Room temperature
Expiry date: According to the sponsor, the stability of the test item is guaranteed throughout the study period when stored under the conditions stated.

Test animals

Species:

rat

Strain:

other: Crl: CD(SD)

Sex:

female

Details on test animals or test system and environmental conditions:

Species, strain: Rats, Crl:CD(SD).
Supplier: Charles River Deutschland GmbH, D-97633 Sulzfeld.
Number and sex: 6 females.
Age: Between 8 and 9 weeks at the time of the administration.
Health conditions: A health inspection was performed prior to the commencement of treatment to ensure that the animals were in a good state of health.
Hygiene: Optimal hygienic conditions.
Room number: EH1-23.
Room temperature: a mean of 20.95 °C (continuous control and recording).
Relative humidity: a mean of 58.53 % (continuous control and recording).
Air exchange: 12 per hour.
Light: Artificial light from 6 a.m. to 6 p.m.
Cages: Single caging in Makrolon cages type III (37.5 cm x 21.5 cm bottom area, 18 cm height). Wire mesh lids. Sanitation of cages once a week.
Bedding material: Aspen wood chips, supplied by ABEDD®, LAB & VET Service GmbH, Vienna, autoclaved. Random samples of the bedding material are analysed for contaminants by the supplier. Changes 1 / week..
Environmental enrichment: Nibbling wood bricks (10 cm x 2 cm x 2 cm) and nesting material, both from the same material and source as the bedding material, were offered to the animals once a week.
Feed: Ssniff R/M-H maintenance diet for rats and mice (item V1534-300) ad libitum, supplied by Ssniff Spezialdiäten GmbH, 59494 Soest, Germany. Analysis of the feed for ingredients and contaminants is performed randomly by Ssniff.
Exception: The feed was withdrawn the evening before the administration of the test substance and was offered again about three hours afterwards.
Water: Tap water from an automatic watering system, ad libitum. Random samples of the water are analysed by the "AGES", A-1226 Vienna, to check, if the water fulfils the requirements for drinking water for humans.
Identification: Labelling with felt-tipped pen on the tail and on the cage.
Acclimatisation: At least 7 days.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

(deionised)

Details on oral exposure:

A peroral administration was performed once in the morning by stomach intubation using a metal gavage.
Vehicle: deionised water.
Dose volume: 20 mL per kg body weight. The individual dose volumes were calculated using the body weights determined on the day of the administration.
Justification: The test substance was not soluble in water, but a homogeneous suspension could be prepared with water and water shall be used preferably, according to the guidelines.

Annex 2d of the OECD guideline and Annex 1d of the EC-Directive were used for the stepwise dosing to groups of 3 females each.
The sequence of dosing of the test substance was:
• Step 1: 2000 mg per kg body weight.
• Step 2: 2000 mg per kg body weight.
A Limit-Test with the starting dose of 2000 mg/kg body weight was requested by the sponsor. The further proceeding was in accordance with the guideline/directive.

Doses:

2000 mg per kg body weight

No. of animals per sex per dose:

3 per step (6 females per dose)

Control animals:

no

Details on study design:

Observations were performed within the periods 0 - 0.5, 0.5 - 1, 1 - 2, 2 - 4 and 4 - 6 hours after administration (p.a.) of the test substance and then at least once a day for a total of 14 days. Observations included but were not limited to changes in skin, fur, eyes, the occurrence of secretions and excretions, autonomic activity, changes in gait, posture and the presence of convulsions.
Body weights were determined
• before administration.
• 7 days p.a.
• 14 days p.a.
Body weight gain was calculated for each week of the study, i.e. between
• 0 and 7 days p.a.
• 7 and 14 days p.a.
The animals were sacrificed by inhalation of 80 % CO2 + 20 % air 14 days p.a. and subjected to a necropsy including a gross pathological examination.

Statistics:

Not applicable

Results and discussion

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Based on the guidance given in OECD guideline 423 (annex 2d), the LD50, oral can be assessed to be > 5000 mg/kg body weight.

Mortality:

No mortality occurred.

Clinical signs:

No toxic effects of the test substance were noted by signs in life and post mortem at a dose of 2000 mg test substance per kg body weight.

Body weight:

5/6 animals gained weight in both weeks p.a., 1/6 animals gained no weight in the second week p.a.

Gross pathology:

No abnormal findings were made in all animals at the necropsy 14 d p.a.

Any other information on results incl. tables

Table: Body weights and body weight gain

          Individual data, means and standard deviations SD.

Dose	Animal	Body weight (g)			Body weight gain (g)
mg/kg (Step No.)	No.	before administr.	7 days p.a.	14 days p.a.	death	0-7 days p.a.	7-14 days p.a.
2000	51	209	247	255	-	38	8
(1)	52	208	227	234	-	19	7
	53	210	238	238	-	28	0
	mean	209.0	237.3	242.3	-	28.3	5.0
	SD	1.0	10.0	11.2	-	9.5	4.4
2000	54	200	235	253	-	35	18
(2)	55	199	237	254	-	38	17
	56	196	231	250	-	35	19
	mean	198.3	234.3	252.3	-	36.0	18.0
	SD	2.1	3.1	2.1	-	1.7	1.0

Table:  Observations in life

Findings	Dose (mg/kg), Step No.	Animal Nos.	Observation time (p.a.) first    last
no clinical signs	2000, 1	51, 52, 53	0 h / 14 d
no clinical signs	2000, 2	54, 55, 56	0 h / 14 d

 

Table: Necropsy findings

          Number of animals examined: 6 females.

SYSTEM Organ, finding	Dose (mg/kg)	Step No.	Animal Nos.
no abnormal findings	2000	1	51, 52, 53
no abnormal findings	2000	2	54, 55, 56

 

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to Commission Directive 2001/59/EC "DOPO-OX-Ammonium" does not require classification for acute oral toxicity.

Executive summary:

Methods and investigations were performed in conformance with the OECD-Guideline 423, 17 December 2001 and the Council Regulation (EC) No 440/2008,Method B.1 tris.

Administration of the test substance

"DOPO-OX-Ammonium" was administered once orally via gavage as a suspension in deionised water to female Crl:CD(SD) rats.

The dosing was performed sequentially to groups of 3 animals per step using a starting dose of 2000 mg per kg body weight. The dose volume was 20 mL per kg body weight for all groups.

Investigations

·     Body weights: before administration, 7 and 14 days after the administration (p.a.).

·     Clinical observations: at least once per day.

·     Necropsy: The animals were sacrificed and necropsied 14 days p.a.

Results

Dose (mg/kg)	Step No.	No. of animals			Prominent findings
		exposed	affected	deceased	in life	post mortem
2000	1	3	0	0	none	none
2000	2	3	0	0	none	none

 

Presence of signs in life	no signs
Full recovery of the animals	not applicable
Body weights	5/6 animals gained weight in both weeks p.a., 1/6 animals gained no weight in the 2^ndweek p.a.
Findings in life and post mortem indicate	no toxic effects present
LD50, oralaccording to OECD	> 5000 mg/kg body weight