Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Acute oral toxicity: Key study. OECD 423. GLP study. The oral LD50 was determined to be between 200-500 mg/kg bw in rats.

Acute dermal toxicity: Key study. OECD 402 (Fixed Dose Procedure), GLP study. The dermal LD50 was determined to be between 2000-5000 mg/kg bw in rats.

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:
July 2, 2001 - July 16, 2001
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
Doses: 200 and 500 mg/kg bw
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan, Ibérica S.A., Sant Feliu de Codines, Barcelona
- Females (if applicable) nulliparous and non-pregnant: Yes
- Weight at study initiation: Male: 156-211 g, Females: 149-170 g
- Fasting period before study: 18-20 hours before administration of selected doses and after 2 horas, the diet was administrated again.
- Housing: Makrolon cages with a basal surface of 100 cm2, containing a chip Lignocel S 8-15 on the floor to detect possible contaminants. Every cage contained max. 5 rats of the same sex.
- Diet (e.g. ad libitum): ad libitum, Harlan, Ibérica S.A., (diet for rat - mouse maintenance RMM, lot. batch: URTG-00240010613, expiry date: 13/09/2001), that has been analyzed by the manufacturer to detect possible contaminants.
- Water (e.g. ad libitum): ad libitum. Periodically, microbiological controls were carried out on the water, supplied by the Barcelona Water Company, which is offered to animals through bottles.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: June 27, 2001 - July 16, 2001
Route of administration:
oral: unspecified
Vehicle:
sorbitan derivative
Remarks:
aqueous solution of TWEEN 80 at 1%
Details on oral exposure:
VEHICLE
- Concentration in vehicle:10 mg/mL and 25 mg/mL

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg

DOSAGE PREPARATION (if unusual): The suspensions of the product were prepared at concentrations 10 mg/mL and 25 mg/mL using as a suspending agent an aqueous solution of TWEEN 80 at 1%

CLASS METHOD (if applicable)
Rationale for the selection of the starting dose: Not available.
Doses:
200 and 500 mg/kg bw
No. of animals per sex per dose:
3 rats per sex and dose.
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Observations: Twice on the day of administration and once daily after administration.
Weighing: Before administration and on days 7 and 14 after administration.
- Necropsy of survivors performed: Yes.
After 14 days of observation after treatment, the animals were sacrificed by one injection of 1 mL of pentobarbital 0.2 g/mL (Dolethal R). Immediately the necropsy of all the animals was performed. Tissues were observed "in situ" after opening of the thoracic and abdominal capitula. The location, colour, appearance, size of the tissues, organs and any anomality were noted. The animals that died before the end of the observation period were also subject to necropsy.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
>= 200 - <= 500 mg/kg bw
Based on:
test mat.
Mortality:
200 mg/kg: No death was observed.
500 mg/kg: One male died before finalization of the first hour of observation and 3 females died before 4 hours of observation. Two males have been found dead at 24 hours of observation.
Clinical signs:
200 mg/kg: One female showed up anti-luteal posture at the time of administration. At 4 hours all the animals had signs of diarrhea and after 24 hours- mydriasis. All these signs reversed before 24 hours

500 mg/kg: Immediately after administration, all the animals observed a decrease in motor activity, post-traumatic stress and diarrhea in half of the animals. The three females also had salivation. The males that survived after the first hour after administration maintained the previous signs and also mydriasis was observed. The females presented ataxia, prostration, lacrimation, anti-luteal posture and one of them respiratory difficulty after one hour of being administered.
Body weight:
The body weight of the animals at a dose of 200 mg/kg evolved within the margins established for their strain, sex, and age.
Gross pathology:
200 mg/kg: The observation of the surviving animals administered at a dose of 200 mg/kg was normal, although there was a linear ulcer of the stomach of 1 mm and a female had a slightly congested intestine.

500 mg/kg: Males that were found dead at 24 hours after administration had the following signs at necropsy: Dorsal curvature; amber intestinal content, in one of them the content of the caecum was liquid ulcerated stomach, in one of them they found hemorrhage; liver slightly darkened; loss of consistency of the spleen, stomach, intestine, seminal vesicles; lungs and pooled thymus and hardened heart. The male which died in the first hour after administration showed hemorrhagic spots and a 3 mm ulcer in the stomach. Moreover, the gastric mucosa was pale and the liver was slightly darkened. The females showed intestinal content of amber hue; presence of hemorrhagic points or small ulcers in the stomach; dark liver; congested lungs and slightly hardened heart.

Table 1: Results:

Doses (mg/kg)

Animals

In life

Dead

% Mortality

200

3 males

3

0

0

3 females

3

0

0

500

3 males

0

3

100

3 females

0

3

100

Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
The oral LD50 was determined to be between 200-500 mg/kg bw in rats.
Executive summary:

The acute oral toxicity of the test item has been tested in accordance with OECD Guideline 423 (GLP study). The test item was administered by oral route to a group of 12 Sprague-Dawley rats (3 rats per sex and dose) at 200 and 500 mg/kg bw. The rats were observed during 14 days after administration. No mortality was observed at the dose of 200 mg/kg bw. The dose produced signs of diarrhea during the first hour and mydriasis the day after administration. All of the animals recovered. The necropsy of these animals did not reveal any anomaly. All animals exposed to a dose of 500 mg/kg bw died during the first 24 hours of observation. The clinical observations were decreased motor activity, diarrhea and prostration accompanied by respiratory difficulties. Mydriasis was observed in the males that did not die during the first hour and ataxia, salivation or lacrimation in the females. The necropsy revealed mainly gastrointestinal effects, such as ulcerations and gastric hemorrhages. Based on these results, the oral LD50 in rats was determined to be between 200 and 500 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
200 mg/kg bw

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:
March 27, 2019 - April 10, 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
Oct 2017
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-house bred animals (Bioneeds India Private Ltd., Devarahosahalli Sompura Hobli, Taluk, Nelamangala, Karnataka 562111, India)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 wk
- Weight at study initiation: 218.43 - 225.54 g
- Housing: A maximum of three animals were housed in a standard polypropylene cage (size: L 430 x B 285 x H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube. For range finding study, animals were housed individually after treatment throughout the observation. For the main study, during treatment, the animals were housed individually and after patch removal they were housed together. Clean sterilized paddy husk was provided as bedding material. Paper shredding was provided as enrichment.
- Diet: Altromin maintenance diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG) was provided ad libitum.
- Water: Deep bore-well water passed through Reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes, ad libitum.
- Acclimation period: yes.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3 - 22.8°C
- Humidity (%): 42 - 65 %
- Air changes (per hr): 12 - 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 27/03/2019 To: 22/04/2019 (depending on the animal)
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: approximately 6 cm × 10 cm, on the dorso-lateral area of the trunk of the animals
- % coverage: at least 10% of the body surface area
- Type of wrap if used: The porous gauze dressing was then wrapped with non-irritating adhesive tape and finally, the application site was wrapped using crepe bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes. At the end of the contact period, the residual test item was washed with distilled water and dried with absorbent cotton.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- For solids, paste formed: yes. The required quantity of the test item per animal (based on the individual animal day 1 body weight) was weighed on an aluminium foil and moistened with 0.5 mL of distilled water to form a thin paste using glass rod.
Duration of exposure:
24 h
Doses:
- Range finding study: 200, 1000, 2000 mg/kg bw.
- Main study: 2000 mg/kg bw.
No. of animals per sex per dose:
1 female per dose in the preliminary study; 2 females per dose in the main test.
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All the animals were observed for clinical signs of toxicity and mortality at 20 to 30 min, 1 hr (±10 mins), 2 hrs (±10 mins), 4 hrs (±10 mins) and 6 hrs (±10 mins) on treatment day 1 and thereafter once daily for clinical signs of toxicity and twice daily for mortality during the 14 days observation period. The body weight was recorded on day 1 before test item application and on day 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.
Preliminary study:
No clinical signs or mortalities were observed with any animal at any dose tested (200, 1000 or 2000 mg/kg bw).
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 2 000 - <= 5 000 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 was observed throughout the study.
Clinical signs:
No clinical signs or skin reactions were noted throughout the study.
Body weight:
No treatment related changes in body weight and percent change in body weight with respect to day 1 were noted.
Gross pathology:
No treatment related gross pathological changes were noted in any of the dosed animals (range finding study and main study) during necropsy.

Table 1. Individual animal clinical signs of toxicity and mortality record.

Phase of the Experiment

 

Dose (mg/kg body weight)

Animal No.

Sex

Time of Dosing

(AM)

Clinical Signs of Toxicity and Mortality on Day 1

Clinical Signs of Toxicity and Mortality on days

20-30

mins

1 hr

(±10 mins)

2 hrs

(±10 mins)

4 hrs

(±10 mins)

6 hrs

(±10 mins)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Range Finding Study

200

Rd5843

F

11:22

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

1000

Rd5844

F

10:42

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

2000

Rd5845

F

10:24

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Main Study

2000

Rd5846

F

11:40

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd5847

F

11:41

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Table 2. Individual animal body weights (g) and % changes in body weight with respect to day 1.

Phase of the Experiment

Dose (mg/kg)

Animal No.

Sex

Body Weight (g) on Days

Percent Change in Body Weight with Respect to Day

1

8

15

1 to 8

1 to 15

Range Finding Study

200

Rd5843

F

235.77

247.52

260.15

 

4.98

10.34

1000

Rd5844

F

239.21

251.89

267.58

 

5.30

11.86

2000

Rd5845

F

247.79

261.53

273.97

 

5.55

10.57

Main Study

2000

Rd5846

F

243.90

256.19

270.93

 

5.04

11.08

Rd5847

F

241.21

258.83

272.05

7.30

12.79

Mean

 

242.56

257.51

271.49

 

6.17

11.93

±SD

 

1.90

1.87

0.79

 

1.60

1.20

n

 

2

2

2

 

2

2

Table 3. Individual animal gross pathology findings.

Phase of the Experiment

Dose

(mg/kg body weight)

Animal No.

Sex

Fate

Gross Pathology Findings

External

Internal

Range finding Study

200

Rd5843

F

TS

NAD

NAD

1000

Rd5844

F

TS

NAD

NAD

2000

Rd5845

F

TS

NAD

NAD

Main Study

2000

Rd5846

F

TS

NAD

NAD

Rd5847

F

TS

NAD

NAD

Interpretation of results:
GHS criteria not met
Remarks:
EU criteria
Conclusions:
The dermal LD50 was determined to be between 2000-5000 mg/kg bw in rats.
Executive summary:

An acute dermal toxicity study of the test item in rats was performed, according to OECD 402 (Fixed Dose Procedure), under GLP conditions. In the range-finding study, the test item was applied onto the intact skin of one female Sprague Dawley rat per dose in a stepwise procedure, at doses of 200 (starting dose), 1000 and 2000 mg/kg bw, respectively. Based on the results of the range-finding study, where no clinical signs or mortalities were observed, a main study was performed with 2 further rats at 2000 mg/kg bw test item. All the animals were observed for clinical signs of toxicity and mortality at 30 min, 1, 2, 4, and 6 hrs (±10 mins) on treatment and daily thereafter during the 14 days observation period. Body weights were recorded before test item application, and on days 8 and 15. At the end of observation period, all the animals were sacrificed under carbon dioxide asphyxiation and subjected to necropsy and detailed gross pathological examination. No mortality, clinical signs, skin reactions, treatment related body weight changes or gross pathological changes were observed at any of the dose steps. Therefore, it was concluded that the acute dermal median lethal dose of test item in rats is > 2000 mg/kg body weight, resulting in 2000 < LD50 ≤ 5000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
One key study of Klimisch 1 is available.

Additional information

Justification for classification or non-classification

Based on the available information (the value of LD50 evaluated between 200 - 500 mg/kg bw), the substance is classified for Acute (oral) toxicity Category 3 (H301), according to CLP Regulation (EC) no. 1272/2008.

Categories Display