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Diss Factsheets

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 10, 2017 - May 24, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
calcium tetrahydrate chloride [2-(trimethylazaniumyl)ethyl] phosphate
Cas Number:
72556-74-2
Molecular formula:
C5H21CaClNO8P
IUPAC Name:
calcium tetrahydrate chloride [2-(trimethylazaniumyl)ethyl] phosphate
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
SPF Caw
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: JANVIER LABS (53940 Legenest St. Isle - France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: males: 7 weeks old; females: 8 weeks old
- Weight at study initiation: males: 257.4 ± 9.1; females: 214.2 ± 5.7
- Fasting period before study: overnight
- Housing: Groups of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet (e.g. ad libitum): Foodstuff (ENVIGO -2016) ad libitum
- Water (e.g. ad libitum): Tap-water from public distribution system ad libitum. Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas- Eurofins (France)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC - 25ºC. A temperature lower than 19ºC was registered on 18 of May 2017. The minimum value measured was 27ºC.
- Humidity (%): 30% - 70%. A relative humidity lower than 30% was registered on 10, 13, 16, 17, 23 and 24 of May 2017. The minimum value measured was 18%.
- Air changes (per hr): 10 changes/hour
- Photoperiod (hrs dark / hrs light): 12 h light (7 - 19 h) / 12 h darkness

IN-LIFE DATES: From: 10 May 2017 To: 24 May 2017

Administration / exposure

Type of coverage:
other: non-occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal area of the trunk of the animal
- % coverage: 10% of the body surfade area
- Type of wrap if used: non-occlusive porous gauze dressing (50 mm x 50 mm non woven swab, 4-layer patch, MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic micropore TM adhesive tape from 3M)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): after removing the gauze dressings, the treated area was rinsed with destilled water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
- Concentration (if solution): 0.2 g/ml
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- Amount(s) applied (volume or weight with unit): 10 mL/Kg bw
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
yes
Remarks:
Study No. TAD-2016-004 (see "Other information on results").
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily observations, the animals were weighed on day 0 (just before administering the test item) then on day 2, day 7, and day 14.
- Necropsy of survivors performed: yes.
At the end of the study, the animals were euthanized with sodium pentobarbital (Dolethal®). Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.
Gross pathology: On D14, the animals were anaesthetised with sodium pentobarbital and administration continued to fatal levels. Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. Parameters examined: Oesophagus, Stomach, Duodenum, Jejunum, Ileon, Caecum, Colon, Rectum, Spleen, Liver, Thymus, Trachea, Lungs, Heart, Kidneys, Urinary Bladder, Ovaries, Uterus, Treatment Area, Adrenals and Pancreas.
- Other examinations performed:
Clinical signs: Spontaneous activity, Preyer's reflex (noise), Respiratory rate, Convulsions, Tremors, Body temperature, Muscle tone, Palpebral opening, Pupil appearance, Salivation, Lachrymation, Righting reflex, Treatment site, Mortality.

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 during the study.
Clinical signs:
Neither cutaneous reactions nor systemic clinical signs related to the administration of the test item were observed.
Body weight:
The body weight evolution of the animals remained normal throughout the study.
Gross pathology:
The macroscopic examination of the animals at the end of the study did not reveal treatment-related changes.

Any other information on results incl. tables

Table 1. Test item at 2000 mg/kg bw. Body weight and weight gain in grams.

MALES

D0

D2

D2-D0

D7

D7-D0

D14

D14-D0

Rm 1455

261

266

5

310

49

354

93

Rm 1456

255

261

6

313

58

360

105

Rm 1457

258

262

4

311

53

348

90

Rm 1458

244

256

12

298

54

344

100

Rm 1459

269

280

11

318

49

369

100

MEAN

257.4

265.0

7.6

310.0

52.6

355.0

97.6

Standard deviation

9.1

9.1

3.6

7.4

3.8

9.9

6.0

FEMALES

D0

D2

D2-D0

D7

D7-D0

D14

D14-D0

Rf 1460

211

216

5

231

20

300

89

Rf1461

220

218

-2

238

18

269

49

Rf 1462

218

219

1

240

22

264

46

Rf 1463

206

212

6

216

10

248

42

Rf 1464

216

228

12

239

23

256

40

MEAN

214.2

218.6

4.4

232.8

18.6

267.4

53.2

Standard deviation

5.7

5.9

5.3

10.0

5.2

19.9

20.3

GENERAL APPEARANCE BEFORE AUTOPSY: Normal

Observations: Nothing to report.

Current control Study:

The study identified No. TAD -2016 -004 was performed to assess the comportament of the strain of rat used at the laboratory in the environment and to give additional historical data. The method was designed to meet the requirements of the following: OECD Guideline No. 402 (February 24th, 1987) and Method B.3 of Council regulation No. 440/2008.

 

Test item : Distilled water

Study dates: 20 September 2016 to 04 October 2016

Results:

- Clinical examinations: Nothing to report. Animal normal (10/10)

- Bodyweight evolution: normal during the test.

- Necropsy: No treatment related changes were observed.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 of the test item in rats is greater than 2000 mg/kg bw.
Executive summary:

A limit test was performed to determine the acute dermal toxicity of the test item in rats according to OECD 402 and EU method B.3, following GLP. The test item was applied onto the intact skin of 10 Sprague Dawley rats (5 males, 5 females) at a dose of 2000 mg/kg bw. All animals were observed once daily for 14 days, survival and body weight were monitored. There were no mortality and systemic clinical signs related to the administration of the test item during the study. Neither cutaneous reactions nor systemic clinical signs related to the administration of the test item were observed. Normal changes in body weights and none macroscopic findings were found. In conclusion, the test item was found to be non toxic by dermal route, with an LD50 > 2000 mg/kg bw in rats.