Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-07-21 - 2017-11-17
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)
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Qualifier:
according to guideline
Guideline:
other: Japan MAFF Testing Guideline of 12 Nousan No. 8147
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes

Test material

Constituent 1
Reference substance name:
Reaction mass of Disodium (sulphonatothio)acetate and sodium chloride
EC Number:
947-115-0
Molecular formula:
C2H2Na2O5S2.NaCl
IUPAC Name:
Reaction mass of Disodium (sulphonatothio)acetate and sodium chloride
Test material form:
liquid
Details on test material:
Purity of Test material:
60.9 % Disodium (sulphonatothio)acetate
20.3 % NaCl
4.3 % Na2S2O3
0.6 % NasSO4
5.0 % H2O
Specific details on test material used for the study:
Analysis of the test item was carried out at BASF SE, Germany.

Information on stability or characterization of the test item, which appropriately defines the test batch, is under the responsibility of the sponsor.

Name of test item: Disodium (sulphonatothio)acetate; dried
Test item No.: 14/0318-2
Batch identification: Sample 14/078 aus Mollescal HW, charge 10924344R0
Content: 60.9 g/100 g test item; 20.3 g/100 g NaCl
Determined by 1H-NMR spectroscopy.
(For details see analytical report No. 15L00094)
Homogeneity: The test item was homogeneous by visual inspection.
Storage stability: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
Expiry date: April 28, 2019
Storage conditions: Room temperature
Physical state/ color: Solid / white

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Age on day 0: Young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks). The female animals were nulliparous and non-pregnant.
Supplier: Charles River Wiga GmbH, Germany
Arrival in the testing facility: Acclimatization period of at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.
Identification: Individual identification by cage cards and tail marking.
Body weight on day 0: Animals of comparable weight (± 20% of the mean weight, actual weights)
Room temperature / relative humidity: The animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 22°C  3°C for temperature and of 30 – 70% for relative humidity. There were no deviations from these ranges, which influenced the results of the study.
Air changes per hour: Approx. 10
Day / night rhythm: 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)
Type of cage: Makrolon cage, type III
Number of animals per cage: Single housing
Feeding: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
Drinking water: Tap water ad libitum
Bedding: H 15005-29; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)
Enrichment: Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
Clipping of the fur: About 24 hours before application
Route of application: Single application to the clipped skin (dorsal and dorsolateral parts of the trunk); covering of the application site with a semi-occlusive dressing for 24 hours. Afterwards removal of the semi-occlusive dressing, rinsing of the application site with warm water.
Application area: About 40 cm² (corresponds to at least 10% of the body surface)
Time of day of application: In the morning
Duration of exposure:
24 h
Doses:
1000 and 2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females, respectively) were dermally exposed to doses of 2000 mg/kg and 1000 mg/kg bw of Disodium (sulphonatothio)acetate; dried (as solution in deionized water). The clipped application site (dorsal and dorso-lateral parts of the trunk, comprising at least 10% of the total body surface) was covered by semi-occlusive dressing during the 24- hour exposure period. The animals were observed for 14 days.

Body weight determination: Individual body weights shortly before application (day 0), weekly thereafter and on the last day of observation.
Clinical observations: Clinical signs for each animal were recorded several times on the day of application and at least once during each workday thereafter.
Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation.
Mortality: A check for any dead or moribund animals was made at least once each workday, these records are archived by Bioassay.
Pathology: Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually
increasing concentrations.
The evaluation of skin reactions was performed according to Draize, J. H. "Dermal toxicity." Appraisal of the safety of chemicals in foods, drugs and cosmetics (1959): 46-59. Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas:

Erythema and eschar formation

Grading
0: No erythema
1: Very slight erythema (barely perceptible)
2: Well- defined erythema
3: Moderate to severe erythema
4: Severe erythema (beet redness) to eschar formation preventing grading of erythema


Edema formation

Grading
0: No edema
1: Very slight edema (barely perceptible)
2: Slight edema (edges of area well- defined by definite raising)
3: Moderate edema (raised approx. 1 mm)
4: Severe edema (raised more than 1 mm and extending beyond area of exposure)
Statistics:
Calculations were performed using Microsoft Excel 2010 and checked with a calculator.

Results and discussion

Preliminary study:
By request of the sponsor a starting dose of 1000 mg/kg bw was chosen in the first step with 5 male and 5 female animals. As no mortality occurred and no local skin findings were observed, a dose of 2000 mg/kg bw was applied to another group of 5 male and 5 female rats in the second step.
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
Clinical signs:
No systemic clinical signs were observed in both dose groups (2000 mg/kg bw and 1000 mg/kg bw) during clinical examination.
No local skin effects were observed in both dose groups (2000 mg/kg bw and 1000 mg/kg bw.) during clinical examination.
Body weight:
The body weight of all male animals in both test groups increased within the normal range throughout the study period.
The body weights of the female animals of the 2000mg/kg bw test group increased in two animals within the normal range throughout the study period. The three other females showed a normal body weight increase during the first week, but a stagnation or marginal loss of body weight during the second week.
Four female animals of the 1000 mg/kg bw test group showed a loss of body weight during the first week, but a normal body weight increase during the second week, while in the fifth animal a stagnation of body weight was observed during the whole observation period.
Due to the fact that stagnation or slight loss of body weight is commonly known for females after dermal applications, the observed stagnation or marginal loss of weight is considered to be unspecific.
Gross pathology:
No macroscopic pathologic abnormalities were noted in any animal examined on the last day of observation (2000 mg/kg bw.: 5 males and 5 females; 1000 mg/kg bw.: 5 males and 5 females).

Any other information on results incl. tables

MORTALITY

 

Mortality

Dose (mg/kg bw):

2000

2000

Sex:

male

female

Application:

1

1

No. of animals:

5

5

Mortality (animals):

-

-

 

Mortality

Dose (mg/kg bw):

1000

1000

Sex:

male

female

Application:

1

1

No. of animals:

5

5

Mortality (animals):

-

-


 

 

 

 

MAXIMUM INCIDENCE OF CLINICAL SIGNS

 

Maximum incidence of systemic clinical signs (males)

Dose (mg/kg bw):

2000

Sex:

male

Application:

1

No. of animals:

5

Animal No.:

R476

R477

R478

R479

R480

Abnormalities:

-

-

-

-

-

 

Maximum incidence of systemic clinical signs (females)

Dose (mg/kg bw):

2000

Sex:

female

Application:

1

No. of animals:

5

Animal No.:

R481

R482

R483

R484

R485

Abnormalities:

-

-

-

-

-

 

Maximum incidence of systemic clinical signs (males)

Dose (mg/kg bw):

1000

Sex:

male

Application:

1

No. of animals:

5

Animal No.:

R441

R442

R443

R444

R445

Abnormalities:

-

-

-

-

-

 

 

Maximum incidence of systemic clinical signs (females)

Dose (mg/kg bw):

1000

Sex:

female

Application:

1

No. of animals:

5

Animal No.:

R446

R447

R448

R449

R450

Abnormalities:

-

-

-

-

-


 

 

NATURE AND DURATION OF LOCAL CLINICAL SIGNS

 

Nature and duration of local clinical signs(males)

Dose (mg/kg bw):

2000

Sex:

male

Application:

1

No. of animals:

5

Animal No.:

R476

R477

R478

R479

R480

Abnormalities:

-

-

-

-

-

 

Nature and duration of local clinical signs(females)

Dose (mg/kg bw):

2000

Sex:

female

Application:

1

No. of animals:

5

Animal No.:

R481

R482

R483

R484

R485

Abnormalities:

-

-

-

-

-

 

MNature and duration of local clinical signs(males)

Dose (mg/kg bw):

1000

Sex:

male

Application:

1

No. of animals:

5

Animal No.:

R441

R442

R443

R444

R445

Abnormalities:

-

-

-

-

-

 

 

Nature and duration of local clinical signs(females)

Dose (mg/kg bw):

1000

Sex:

female

Application:

1

No. of animals:

5

Animal No.:

R446

R447

R448

R449

R450

Abnormalities:

-

-

-

-

-


 

 

 

BODY WEIGHTS

 

Individual body weight changes

Dose (mg/kg bw):

2000

Sex:

male

Administration:

1

Animal No.:

R

R

R

R

R

Mean

weight

Standard-

deviation

476

477

478

479

480

Body weight at

study day (g):

 

 

 

 

 

 

 

0

227

227

225

234

239

230.4

5.90

7

256

265

250

256

263

258.0

6.04

14

272

287

276

280

289

280.8

7.19

 

 

 

 

 

 

 

 

Individual body weights changes

Dose (mg/kg bw):

2000

Sex:

female

Administration:

1

Animal No.:

R

R

R

R

R

Mean

weight

Standard-

deviation

481

482

483

484

485

Body weight at

study day (g):

 

 

 

 

 

 

 

0

209

210

206

212

209

209.2

2.17

7

216

220

216

219

220

218.2

2.05

14

214

223

221

224

220

220.4

3.91

 

Individual body weight changes

Dose (mg/kg bw):

1000

Sex:

male

Administration:

1

Animal No.:

R

R

R

R

R

Mean

weight

Standard-

deviation

441

442

443

444

445

Body weight at

study day (g):

 

 

 

 

 

 

 

0

239

231

233

222

244

233.8

8.35

7

266

250

256

237

272

256.2

13.72

14

301

271

281

257

310

284.0

21.63

 

 

 

 

 

 

 

 

Individual body weights changes

Dose (mg/kg bw):

1000

Sex:

female

Administration:

1

Animal No.:

R

R

R

R

R

Mean

weight

Standard-

deviation

446

447

448

449

450

Body weight at

study day (g):

 

 

 

 

 

 

 

0

209

208

219

211

214

212.2

4.44

7

206

208

207

210

203

206.8

2.59

14

214

212

222

221

216

217.0

4.36


 

 

 

 

PATHOLOGY

 

Gross Pathology

Dose (mg/kg bw):

2000

Sex:

male

Application:

1

No. of animals:

5

Animal No.:

R476

R477

R478

R479

R480

Macroscopic pathologic abnormalities :

-

-

-

-

-

 

 

Gross Pathology

Dose (mg/kg bw):

2000

Sex:

female

Application:

1

No. of animals:

5

Animal No.:

R481

R482

R483

R484

R485

Macroscopic pathologic abnormalities :

-

-

-

-

-

 

Gross Pathology

Dose (mg/kg bw):

1000

Sex:

male

Application:

1

No. of animals:

5

Animal No.:

R441

R442

R443

R444

R445

Macroscopic pathologic abnormalities :

-

-

-

-

-

 

 

Gross Pathology

Dose (mg/kg bw):

1000

Sex:

female

Application:

1

No. of animals:

5

Animal No.:

R446

R447

R448

R449

R450

Macroscopic pathologic abnormalities :

-

-

-

-

-

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
Under the conditions of this study the median lethal dose (LD50) of Disodium (sulphonatothio)acetate; dried after dermal application was found to be greater than 2000 mg/kg bw in male and female rats.
Executive summary:

In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females, respectively) were dermally exposed to doses of 2000 mg/kg and 1000 mg/kg bw ofDisodium (sulphonatothio)acetate;dried(as solution in deionized water). The clipped application site (dorsal and dorso-lateral parts of the trunk, comprising at least 10% of the total body surface) was covered by semi-occlusive dressing during the 24- hour exposure period. The animals were observed for 14 days. No mortality occurred in the 2000 mg/kg bw. and 1000 mg/kg bw. dose groups. Neither signs of systemic toxicity nor local skin effects were observed in any animal. No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study. The body weight of all male animals in both test groups increased within the normal range throughout the study period. The body weights of the female animals of the 2000 mg/kg bw test group increased in two animals within the normal range throughout the study period. The three other females showed a normal body weight increase during the first week, but a stagnation or marginal loss of body weight during the second week. Four female animals of the 1000 mg/kg bw test group showed a loss of body weight during the first week, but a normal body weight increase during the second week, while in the fifth animal a stagnation of body weight was observed during the whole observation period. Due to the fact that stagnation or slight loss of body weight is commonly known for females after dermal applications, the observed stagnation or marginal loss of weight is considered to be unspecific.

 

Accordingly, the acute dermal median lethal dose (LD50) was determined to be 

LD50, dermal, rat > 2000 mg/kg bw