N,N'-bis(5,5-dimethyl-1,3,2-dioxaphosphinane-2-oxide-2-yl)ethane-1,2-diamine

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 March 2015 to 02 April 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

OECD Guidelines for Testing of Chemicals, Section 4, Number 402 "Acute Dermal Toxicity", adopted February 24, 1987.

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Commission Regulation (EC) No 440/2008, B.3 (L 142, 30 May 2008)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-({2-[(5,5-dimethyl-2-oxo-1,3,2λ⁵-dioxaphosphinan-2-yl)amino]ethyl}amino)-5,5-dimethyl-1,3,2λ⁵-dioxaphosphinan-2-one

EC Number:

835-272-7

Cas Number:

256374-76-2

Molecular formula:

C12H26N2O6P2

IUPAC Name:

2-({2-[(5,5-dimethyl-2-oxo-1,3,2λ⁵-dioxaphosphinan-2-yl)amino]ethyl}amino)-5,5-dimethyl-1,3,2λ⁵-dioxaphosphinan-2-one

Test material form:

solid: particulate/powder

Details on test material:

Name
Name: Reaction products of ethane-1,2-diamine, phosphoryl=trichloride and 2,2-dimethylpropane-1,3-diol which makes N,N'-bis(5,5-dimethyl-1,3,2-dioxaphosphinane=2-oxide-2-yl)ethane-1,2-diamine as a main component
Other name: SH-0850
CAS number: 256374-76-2 (main component)

Structural formula
Molecular formula: C12H26N2O6P2 (main component)
Molecular weight: 356.29 (main component)

Provided sample
Purity of the test substance: 100%
Lot number: SK-241002

Physical-chemical properties
Solubility in water: Less than 0.03% (w/w) by visual observation
Melting point: 277 °C
Appearance at normal temperatures: White powder

Storage condition
The test substance was stored in a dark place at room temperature.

Precaution for handling
Protective gloves, mask, glasses and clothes were put on in order to avoid contacts with skin or eyes and inhalation.

Specific details on test material used for the study:

No further details specified in the study report.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals
Species and strain: Crl:WI rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Justification of strain: Recognized by international guidelines as a recommended test system
Number of animals: 5 animals / sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at dosing: Young adult rats
Body weight range at dosing: Males: 242-262 g; Females: 216-237 g
Age at dosing: Males: 7 weeks; Females: 8-9 weeks
Acclimatization time: 7 days

Husbandry
Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room: 242/6
Housing: Individual caging
Cage type: Type II. polypropylene/polycarbonate
Bedding: “Grade 5” Bedding for Laboratory Animals was available to animals during the study. A copy of the Certificate of Analysis is retained in the archives at CiToxLAB Hungary Ltd.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 21.0 – 23.4 °C
Relative humidity: 31 – 51 %
Ventilation: 15-20 air exchanges per hour
Enrichment: Rodents are housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
The temperature and relative humidity were recorded twice daily during the study.

Food and water supply
Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494, Soest, Germany (Batch no.: 680 2237, Expiry date: March 2015 or Batch no.: 814 3108, Expiry date: August 2015), ad libitum, and tap water from municipal supply, as for human consumption from 500 ml bottles, ad libitum. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Identification
Animals were identified by numbers written on the tail with an indelible marker. The numbers were given on the basis of the CiToxLAB Hungary Ltd. master file, for each animal allocated to the study.
The boxes were identified by cards holding information on the study code, the sex of animals, the dose group, the cage number and the individual animal number.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

The test item was administered in a single dose as supplied by the Sponsor, but sufficient water was used to moisten the test material to ensure good contact with the skin.
A single administration was performed by the dermal route and was followed by a fourteen-day observation period. The test item was applied as supplied.
The back of the animals were shorn (approximately 10% area of the total body surface) approximately 24 hours prior to the treatment. Only the animals without injury or irritation on the skin were used in the test.
On the test day (Day 0), the test item was applied as a single dose of 2000 mg/kg bw after the moistening with sufficient water, applied uniformly over the skin by use of a gauze pad (ca. 5 cm x 5 cm), and remained on the skin throughout a 24-hour exposure period. Sterile gauze pads were placed on the skin of rats at the site of application. These gauze pads were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours. At the end of the exposure period, residual test item was removed, using body temperature water.

Duration of exposure:

24-hour exposure period.

Doses:

A limit dose of 2000 mg/kg bw was chosen by the Study Director in agreement with the Sponsor.

No. of animals per sex per dose:

10 animals (5 males/5 females)/

Control animals:

no

Details on study design:

Clinical observations
A clinical examination was performed on the day of treatment, at 1 and 5 hours after the application of the test item, and once each day for 14 days thereafter.
Observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behaviour pattern. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Skin Irritation
Adverse skin reactions at the site of application were recorded daily following the removal of the dressing (for scoring scheme see "Any other information" for details).

Measurement of body weight
The body weight of all animals was recorded on Day 0 (beginning of the experiment) and on Days 7 and 14.

Post Mortem Investigations
All animals were subjected to gross macroscopic examination. All animals were anaesthetised with an injection of pentobarbital solution and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. Any gross macroscopic findings were recorded.

Statistics:

Not specified

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred after the 24-hour dermal exposure to SH-0850 in Crl:WI rats.

Clinical signs:

other: Systemic Clinical Signs: Each rat was symptom-free during the entire study. Local Dermal Signs: No local dermal signs were recorded after treatment with the test item during the 14 days observation period.

Gross pathology:

No external or internal macroscopic findings were observed at a dose level of 2000 mg/kg bw at necropsy.

Any other information on results incl. tables

Clinical Observations

DOSE LEVEL: 2000 mg/kg bw                                                                              SEX: MALE

Cage No.

Animal No.

Observations

Observations days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

1

2456

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

2

2457

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

3

2458

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

4

2459

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

5

2460

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

 

DOSE LEVEL: 2000 mg/kg bw                                                                              SEX: FEMALE

Cage No.

Animal No.

Observations

Observations days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

6

2461

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

7

2462

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

8

2463

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

9

2464

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

10

2465

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

Remarks:         + = present

                       h = hour(s)       Treatment day = Day 0

                       Frequency of observation = number of occurrence of observation / total number of observations

 

Body Weight and Body Weight Gain

DOSE LEVEL: 2000 mg/kg bw                                                                  SEX: MALE

Cage No.	Animal No.	Boy weight (g) Days			Body Weight Gain (g)
		0	7	14	0-7	7-14	0-14
1	2456	242	285	325	43	40	83
2	2457	259	297	352	38	55	93
3	2458	252	291	336	39	45	84
4	2459	245	289	335	44	46	90
5	2460	262	332	398	70	66	136
Mean:		252.0	298.8	349.2	46.8	50.4	97.2
Standard deviation:		8.6	19.1	28.9	13.2	10.3	22.1

 

DOSE LEVEL: 2000 mg/kg bw                                                                  SEX: FEMALE

Cage No.	Animal No.	Boy weight (g) Days			Body Weight Gain (g)
		0	7	14	0-7	7-14	0-14
6	2461	225	241	264	16	23	39
7	2462	237	229	238	-8	9	1
8	2463	226	231	237	5	6	11
9	2464	234	252	256	18	4	22
10	2465	216	234	239	18	5	23
Mean:		227.6	237.4	246.8	9.8	9.4	19.2
Standard deviation:		8.3	9.3	12.4	11.3	7.8	14.3

 

Necropsy Findings

DOSE LEVEL: 2000 mg/kg bw                                                                                          SEX: MALE

Cage No.	Animal No.	Necropsy Date/ Study Day	External Observations	Internal Observations	Organ/Tissue
1	2456	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
2	2457	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
3	2458	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
4	2459	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
5	2460	02 April 2015 Day 14	No external observations	No internal observations	Not applicable

 

DOSE LEVEL: 2000 mg/kg bw                                                                                          SEX: FEMALE

Cage No.	Animal No.	Necropsy Date/ Study Day	External Observations	Internal Observations	Organ/Tissue
6	2461	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
7	2462	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
8	2463	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
9	2464	02 April 2015 Day 14	No external observations	No internal observations	Not applicable
10	2465	02 April 2015 Day 14	No external observations	No internal observations	Not applicable

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose (LD50) of SH-0850 after a single dermal administration was found to be greater than 2000 mg/kg bw in male and female Crl:WI rats.

Executive summary:

The purpose of the study was to assess the acute dermal toxicity of SH-0850 when administered to rats by a single semi-occlusive dermal application, followed by an observation period of 14 days. The test item was applied dermally to ten (5 males and 5 females) Crl:WI rats as supplied by the Sponsor. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes.

 

Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Rats were euthanized and subjected to a gross macroscopic examination at the end of the 2-week observation period (Day 14).

 

Results

Mortality

No mortality occurred after the 24-hour dermal exposure to SH-0850 in Crl:WI rats.

Systemic clinical signs

Each rat was symptom-free during the entire study.

Local dermal signs

No local dermal signs were recorded after treatment with the test item during the 14 days observation period.

Body weight

Slight body weight loss was observed in a single female animal between Day 0 and Day 7, but it was considered to be incidental and to have no toxicological relevance.

Necropsy

No external or internal macroscopic findings were noted at a dose level of 2000 mg/kg bw at necropsy.

 

Conclusion

The median lethal dose (LD50) of SH-0850 after a single dermal administration was found to be greater than 2000 mg/kg bw in male and female Crl:WI rats.