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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:
Cytidine 3'-(dihydrogen phosphate)
EC Number:
200-556-6
EC Name:
Cytidine 3'-(dihydrogen phosphate)
Cas Number:
63-37-6
Molecular formula:
C9H14N3O8P
IUPAC Name:
cytidine 3'-(dihydrogen phosphate)
Test material form:
solid
Details on test material:
Synonyms: 5-CMP acid, 5'-Cytidylic acid, Cytidine 5'-monophosphate
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: yes

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: Elevage JANVIER LABS (53940 Le Genest St Isle - France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: male animals of the treated group were 7 weeks old and the female were 8 weeks old
- Weight at study initiation: males= 258.8 ± 13, females= 215.4 ± 10.9
- Fasting period before study: no
- Housing: during the treatment, the animals were in individual cages. On D1, the animals were put into their cage by 5. The rats were kept in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains dust free weed shavings which were 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: at least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC - 25ºC
- Humidity (%): 30% - 70% (a temperature lower than 19ºC was registered on 10, 13, 16, 17, 23 and 24 May 2017, the minimum valued measured was 18ºC. A temperature higher than 25ºC was registered on 18 May 2017, the maximum value mesured was 27ºC)
- Air changes (per hr): at least ten changes cycles per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (07:00 to 19:00) and telve hours darkness.

IN-LIFE DATES: From: To: 10 May 2017 - 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 surface area
- Type of wrap if used: non-occlusive porous gauze dressing (50x50 mm2 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 and liquid paraffin
- 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 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males and 5 females per dose
Control animals:
yes
Remarks:
Current control study (TAD-2017-001)
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.
- Other examinations performed:
Body weight and body weight gain: the animals were weighted on D0 (just before administering the test item) then on D2, D7, and D14.
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.
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.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
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 weitght 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

All clinical observations were normal.

Macroscopical examinations: Nothing to report

                       

 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 1465

268

270

2

310

42

334

66

Rm 1466

247

251

4

294

47

348

101

Rm 1467

261

283

22

336

75

359

98

Rm 1468

274

284

10

332

58

369

95

Rm 1469

244

258

14

294

50

329

85

MEAN

258.8

269.2

10.4

313.2

54.4

347.8

89.0

Standard deviation

13.0

14.7

8.0

20.1

12.9

16.7

14.2

FEMALES

D0

D2

D2-D0

D7

D7-D0

D14

D14-D0

Rf 1470

211

219

8

239

28

300

89

Rf 1471

227

229

2

248

21

278

51

Rf 1472

222

226

4

243

21

284

62

Rf 1473

199

203

4

210

11

269

70

Rf 1474

218

219

1

229

11

256

38

MEAN

215.4

219.2

3.8

233.8

18.4

277.4

62.0

Standard deviation

10.9

10.1

2.7

15.0

7.3

16.5

19.3

 

 

Table 6. Necropsy data sheet of males Rm1106 to Rm1110 (28 February 2017)

 

 

Found dead:   

 

 

Euthanasia: X

 

 

At term: X

 

GENERAL APPEARANCE BEFORE AUTOPSY: Normal

 

 

Observed Organs

 

Observations

 

* OESOPHAGUS

X

N.t.R.

* STOMACH

X

N.t.R.

* DUODENUM

X

N.t.R.

* JEJUNUM

X

N.t.R.

* ILEON

X

N.t.R.

* CAECUM

X

N.t.R.

* COLON

X

N.t.R.

* RECTUM

X

N.t.R.

* SPLEEN

X

N.t.R.

* LIVER

X

N.t.R.

* THYMUS

X

N.t.R.

* TRACHEA

X

N.t.R.

* LUNGS

X

N.t.R.

* HEART

X

N.t.R.

* KIDNEYS

X

N.t.R.

* URINARY BLADDER

X

N.t.R.

* OVARIES

X

N.t.R.

* UTERUS

X

N.t.R.

* TREATMENT AREA (Skin)

X

N.t.R

* ADRENALS

X

N.t.R.

* PANCREAS

X

N.t.R.

PARTICULARS. None

 

N.t.R.: Nothing to report

 

 

Table 7. Necropsy data sheet of females Rf1111 to Rm1115 (28 February 2017)

 

 

Found dead:   

 

 

Euthanasia: X

 

 

At term: X

 

GENERAL APPEARANCE BEFORE AUTOPSY: Normal

 

 

Observed Organs

 

Observations

 

* OESOPHAGUS

X

N.t.R.

* STOMACH

X

N.t.R.

* DUODENUM

X

N.t.R.

* JEJUNUM

X

N.t.R.

* ILEON

X

N.t.R.

* CAECUM

X

N.t.R.

* COLON

X

N.t.R.

* RECTUM

X

N.t.R.

* SPLEEN

X

N.t.R.

* LIVER

X

N.t.R.

* THYMUS

X

N.t.R.

* TRACHEA

X

N.t.R.

* LUNGS

X

N.t.R.

* HEART

X

N.t.R.

* KIDNEYS

X

N.t.R.

* URINARY BLADDER

X

N.t.R.

* OVARIES

X

N.t.R.

* UTERUS

X

N.t.R.

* TREATMENT AREA (Skin)

X

N.t.R

* ADRENALS

X

N.t.R.

* PANCREAS

X

N.t.R.

PARTICULARS. None

 

N.t.R.: Nothing to report

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 ans systemic clinical signs related to the administration of the test irem 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.