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Administrative data

Description of key information

- Acute toxicity by oral route: the oral LD50 value of Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in Wistar rats was established to be within the range of 50-300 mg/kg body weight (corresponding to the Active Ingredient).
- Acute toxicity by inhalation route: no data available.
- Acute toxicity by dermal route: the dermal LD50 value of Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in Wistar rats was established to be greater than 2000 mg/kg.

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:
key study
Study period:
from 2015-08-25 to 2015-09-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Principles of method if other than guideline:
Not applicable.
GLP compliance:
yes (incl. QA statement)
Remarks:
2013-05-06
Test type:
acute toxic class method
Limit test:
no
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: Young adult animals (approx. 11 weeks old)
- Weight at study initiation: 172 -214 g
- Fasting period before study: yes
- Housing: 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): at least 10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 25 August 2015 To: 29 September 2015
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
only for 50 mg/kg group
Details on oral exposure:
VEHICLE
- Water: Elix, Millipore S.A.S., Molsheim, France
- Concentration in vehicle:
- Treatment volume:
2000 mg/kg: 2.1157 mL/kg bw.
300 mg/kg: 0.3174 mL/kg bw.
50 mg/kg: 0.5 mL/kg bw.
- Justification for choice of vehicle: the substance is soluble in water

DOSAGE PREPARATION: The preparations (w/w) were kept at room temperature and were dosed within 1 hour after adding the vehicle to the test substance. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies.

CLASS METHOD:
- Rationale for the selection of the starting dose: Toxicity not expected at this dose level
Doses:
First experiment: 2000 mg/kg
Second experiment: 300 mg/kg
Third and fourth experiment: 50 mg/kg
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: day 1: 1h, 2h and 4h after dosing, and then twice daily
- Frequency of weighing: Days 1 (pre-administration), 8 and 15 and at death or necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: macroscopic observations at death or necropsy
Statistics:
Not applicable
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 50 - < 300 mg/kg bw
Based on:
act. ingr.
Mortality:
2000 mg/kg bw: 3/3
300 mg/kg: 3/3
50 mg/kg: 0/6
Clinical signs:
other: At 2000 mg/kg, lethargy, hunched posture, abnormal gait, piloerection and/or ptosis were noted on Day 1. At 300 mg/kg, hunched posture and piloerection were noted on Day 1. At 50 mg/kg, hunched posture and/or piloerection were noted on Day 1.
Gross pathology:
At 2000 and 300 mg/kg, abnormalities of the stomach (irregular surface, hardened and/or watery clear contents (only 300 mg/kg)) were found at macroscopic post-mortem examination.
At 50 mg/kg, no abnormalities were found at macroscopic examination.
(Beginning of autolysis was noted for the animals treated at 50 mg/kg. This was considered not toxicologically relevant).

Body Weights


SEX/DOSE LEVEL


ANIMAL


DAY 1

DAY 2*
body weigth

at death


DAY 8


DAY 15

FEMALES 2000 MG/KG

1

189

181

---

---

2

185

183

---

---

3**

 

 

---

---

MEAN

193

 

 

 

ST.DEV.

11

 

 

 

N

3

 

 

 

FEMALES 300 MG/KG

4**

 

 

---

---

5

192

190

---

---

6

214

211

---

---

MEAN

188

 

 

 

ST.DEV.

28

 

 

 

N

3

 

 

 

FEMALES 50 MG/KG

7

194

na

203

210

8

197

na

212

217

9

177

na

180

189

MEAN

189

 

198

205

ST.DEV.

11

 

17

15

N

3

 

3

3

FEMALES 50 MG/KG

10

184

na

198

211

11

172

na

201

203

12

190

na

208

216

MEAN

182

 

202

210

ST.DEV.

9

 

5

7

N

3

 

3

3

 * Animals 1-6 were found dead on Day 2.

** There were uncertainties in the body weights of these animals (animal 3:205 and 181; animal 4: 158 and 206 gram). These body weights where therefore not used for interpretation. Sufficient data was available.

Na Not applicable

Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
Under the test conditions of this study, the oral LD50 value of Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in Wistar rats was established to be within the range of 50-300 mg/kg body weight (corresponding to the Active Ingredient).
Executive summary:

Assessment of acute oral toxicity with Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in the rat (Acute Toxic Class Method). The study was carried out based on the guidelines described in:

OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method". Commission Regulation (EC) No 440/2008, B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method". EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity" and JMAFF Guidelines (2000), including the most recent revisions.

 

Initially, Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure three additional groups of three females were dosed at 300, 50 and 50 mg/kg body weight as active ingredient. The animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

 

All animals treated at 2000 and 300 mg/kg were found dead on Day 2.

 

At 2000 mg/kg, lethargy, hunched posture, abnormal gait, piloerection and/or ptosis were noted on Day 1.

At 300 mg/kg, hunched posture and piloerection were noted on Day 1.

At 50 mg/kg, hunched posture and/or piloerection were noted on Day 1.

 

The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

 

At 2000 and 300 mg/kg, abnormalities of the stomach (irregular surface, hardened and/or watery clear contents (only 300 mg/kg)) were found at macroscopic post-mortem examination. At 50 mg/kg, no abnormalities were found at macroscopic examination.

 

Under the test conditions of this study, the oral LD50 value of Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in Wistar rats was established to be within the range of 50-300 mg/kg body weight (as active ingredient).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Quality of whole database:
Key study performed according to the OECD 423 guideline study and in compliance with the GLP (Klimisch score = 1).

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 2017-07-11 to 2017-10-17
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
2015-09-22
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany.
- Females (if applicable)female rats were nulliparous and non-pregnant
- Age at study initiation: Young adult rats
- Weight at study initiation: Between 220 g and 245 g
- Fasting period before study:
- Housing: Individual caging Type II. polypropylene/polycarbonate
- Bedding and nesting:“Lignocel 3/4-S Hygienic Animal Bedding” and “Arbocel crinklets natural” nest building material produced by J. Rettenmaier & Söhne GmbH + Co.KG (D-73494 Rosenberg, Germany) was available to animals during the study.
- Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
- Diet (e.g. ad libitum): 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 number: 262 21592, expiry date: 31 January 2018)
- Water (e.g. ad libitum): tap water from the municipal supply, as for human consumption from a 500 mL bottle, ad libitum.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.8 – 28.1 °C (Due to technical reasons, temperature values (maximum of 28.1 °C) outside the expected range of 22 ± 3 °C were recorded during the study. However, these differences of the environmental parameter were considered not to adversely affect the results of or integrity of the study as confirmed by the clinical Veterinarian.)
- Humidity (%): 32 – 70 %
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 2017-07-20 to 2017-08-08
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: back
- % coverage: approximately 10% area of the total body surface
- Type of wrap if used: Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
- Time after start of exposure: 24h

TEST MATERIAL
- The active ingredient content of the test item is 68.8% (w/w), therefore a correction factor of 1.45 was used to calculate the 2000 mg/kg bw dose level.
Duration of exposure:
24h
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males and 5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: 1h, 5h and dailly for 14 days after treatment (Clinical signs). Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.
- Weighing: Days 0, 7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: Macroscopic examination was performed on all animals. After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
act. ingr.
Mortality:
Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer did not cause mortality at the dose level of 2000 mg/kg bw.
Clinical signs:
other: There were no systemic clinical signs noted in any animal throughout the study.
Gross pathology:
At necropsy, a few crusts were seen on the skin at the dorsal thoracic area in one female animal. Besides this, there was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw
Other findings:
None

Clinical observations (2000 mg/kg)

Animal N°

Obervations

Observation days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

M 229

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

M 230

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

M 231

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

M 232

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

M 233

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

F 234

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

F235

Symptom free

+

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

2/16

Crust – treated area

-

-

-

+

+

+

+

+

+

+

+

+

+

+

+

+

13/16

Erythema

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

3/16

F 236

Symptom free

+

+

-

-

-

-

-

-

-

+

+

+

+

+

+

+

9/16

Crust – treated area

-

-

-

+

+

+

+

+

+

-

-

-

-

-

-

-

6/16

Erythema

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

3/16

F 237

Symptom free

+

+

-

-

-

-

+

+

+

+

+

+

+

+

+

+

12/16

Crust – treated area

-

-

-

+

+

+

-

-

-

-

-

-

-

-

-

-

3/16

Erythema

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

2/16

F 238

Symptom free

+

+

-

+

+

+

+

+

+

+

+

+

+

+

+

+

15/16

Erythema

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

1/16

+ = present         h = hour(s)          Treatment day = Day 0                 

- = Absent

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

Erythema severity: 1 = very slight, 2 = well defined; 3 = Modderate to severe; 4 = severe to slight eschar formation

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the acute dermal median lethal dose (LD50 value) of the test item Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer was found to be above 2000 mg/kg bw in male and female Crl:WI rats. There were no systemic effects of treatment.

According to the GHS criteria, Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer can be ranked as Unclassified for acute dermal exposure.
Executive summary:

An acute dermal toxicity study was performed with test item Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer in Crl:WI rats, in compliance with OECD Guideline No. 402.

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as a single dermal 24-hour exposure followed by a 14-day observation period.

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. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

 

RESULTS

Mortality

Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer did not cause mortality at the dose level of 2000 mg/kg bw.

Clinical Observations

There were no systemic clinical signs noted in any animal throughout the study.

Local dermal signs

Very slight erythema in four females (Days 1-3) and crust in three females were seen at the treated area after treatment with the test item. Besides these, no local dermal signs were observed during the 14 days observation period.

Body weight and body weight gain

Body weight gains of Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer treated animals during the study showed no indication of a test item-related effect.

Macroscopic Findings

At necropsy, a few crusts were seen on the skin at the dorsal thoracic area in one female animal. Besides this, there was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

 

CONCLUSIONS

Under the conditions of this study, the acute dermal median lethal dose (LD50 value) of the test item Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer was found to be above 2000 mg/kg bw in male and female Crl:WI rats. There were no systemic effects of treatment.

According to the GHS criteria, Tetrakis[hydroxymethyl]phosphonium sulphate-urea copolymer can be ranked as Unclassified for acute dermal exposure.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
Key study performed according to the OECD 402 guideline study and in compliance with the GLP (Klimisch score = 1).

Additional information

Justification for classification or non-classification

The oral LD50 value of Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in Wistar rats was established to be within the range of 50-300 mg/kg body weight (as active ingredient), (OECD 423, GLP, Kr.1). Therefore, the test substance is considered as toxic if swallowed and classified into category 3 (H301) according to the criteria of the CLP Regulation (EC) N°1272/2008.

The oral LD50 value of Phosphonium, tetrakis (hydroxymethyl)-sulphate (2:1); polymer with urea in Wistar rats was established to be greater than 2000 mg/kg.