Registration Dossier

Administrative data

Description of key information

Testing for acute toxicity gave the following results:

Acute oral toxicity:  LD50: >2000 mg/kg

Acute dermal toxicity: LD50: >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:
08 May 2007 to 23 May 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species and strain: CRL:(WI) BR rats
Source: CHARLES RIVER (EUROPE) LABORATORIES INC. TOXI COOP Ltd. 1103 Budapest, Cserkesz u. 90.
Justification of strain: The Wistar rat as a rodent is one of the standard animal in toxicity studies.
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Number of animals: 6 animals, 3 animals/group
Sex: Female, nulliparous and non-pregnant.
Age of animals: Young healthy adult rats, less than 10 weeks old
Date of receipt: 03 May 2007
Body weight at treatment: Between range of 184 g and 188 g
Acclimatization time: 5 and 6 days (Group 1 and Group 2)
Animal health: Only healthy animals were used for the test. The veterinarian certified healthy status.
Number of animal room: 245-8
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: Laboratory bedding
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 8-12 air exchanges/hour by central air-condition system.
Food and Water Supply: The animals received ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494
Soest Germany, and tap water from municipal supply, as for human consumption from 500 ml bottle ad libitum.
The drinking water is routinely analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Animal Identification The individual identification was performed by numbers on the tail written with a marker pen. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled
Details on oral exposure:
Vehicle
Name: Distilled water
Batch No: 8991006
Expiry Date: October 2009
Supplier: Humantrade Ltd.
Storage: At room temperature

Formulation
For treatment the test item was applied in a concentration of 200 mg/ml in distilled water. Formulations were prepared just before the administration and stirred with a magnetic stirrer up to end of the treatment.
Doses:
Doses
Justification of the dose:
Starting dose was selected on the basis of the information provided by the Sponsor.
The LD50 value was expected to be above 2000 mg/kg bw. A limit test was performed at 2000 mg/kg bw dose.
Three female animals were treated with a dose level of 2000 mg/kg bw of Gelb Sulfato in the first step.
No. of animals per sex per dose:
6 animals all female, 3 animals/group
Control animals:
yes
Details on study design:
A single oral administration - followed by a fourteen-day observation period – was performed by gavage. On the day before each treatment the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before the treatment. The test item was administered by oral gavage in the morning hours. The food was given back 3 hours after the treatment. A constant treatment volume of 10 ml/kg body weight was applied.

Clinical Observations
Careful clinical observation was made 15 and 30 minutes, 1 h, 2 h, 3 h, 4 h, 6 h after the treatment and once each day for 14 days thereafter.
Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern as well. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body Weight Measurement
The body weight was recorded on day 0 (shortly before the treatment), then on days 7 and 14 with precision of 1g.

NECROPSY
Gross necropsy was performed in each experimental animal terminally. Animals were sacrificed by exsanguination under pentobarbital anaesthesia. After the examination of the external appearance the cranial, thoracic and the abdominal cavities were opened, the organs and the tissues were observed.
Abnormalities were recorded on post mortem data sheets.
Statistics:
No data
Preliminary study:
Three female animals were treated with a dose level of 2000 mg/kg bw of Gelb Sulfato in the first step. All animals survived; so three further animals were dosed at 2000 mg/kg bw dose level next day, as the second step. No mortality occurred in either steps, therefore the test was finished meeting the stopping criteria of guidelines (OECD 423, Directive 2004/73/EC B.1.tris, OPPTS 870.1100).
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Gelb Sulfato caused no mortality in female CRL:(WI) BR rats after a single oral (by gavage) administration of 2000 mg/kg bw.
Clinical signs:
Group 1 – 2000 mg/kg bw
Slight activity decrease, hunched back and piloerection were noted for one animal (1/3) two hours after the treatment. There were no test item effect on the physical condition and behaviour of the remaining animals (2/3).
Yellow coloured urine and faces were noted on the day of the treatment and on the next day.

Group 2 – 2000 mg/kg bw
There were no clinical signs on the day of the treatment and the following 14-day observation period. The physical condition and behaviour of animals were considered normal during the whole experiment (3/3).
Yellow coloured urine and faces were noted on the day of the treatment and on the next day.
Body weight:
The mean body weight and the body weight gain of animals were in the normal range during the two weeks observation period, similar to the expected values in untreated animals of the same age and strain.
Gross pathology:
No macroscopic alterations related to the toxic effect of Gelb Sulfato were found at necropsy. Hydrometra (1/3, Group 1) is a common necropsy findings in experimental rats related to the sexual cycle of animals. The pinprick-sized haemorrhages (1/3 in Group 2) observed in the lungs were due to the exsanguination.
Other findings:
None

CLINICAL OBSERVATIONS

 

SUMMARY OF OBSERVATION

Observations

Frequency of symptoms

Group 1

2000 mg/kg bw

Group 2

2000 mg/kg bw

Normal

2/3

3/3

Decreased activity

1/3

0/3

Hunched back

1/3

0/3

Piloerection

1/3

0/3

Comment: Frequency: number of animals with observation / number of animals observed

INDIVIDUAL CLINICAL OBSERVATIONS

Group 1

Animal Number

Observations

Day 0

Day 1-14

F

15’

30’

1h

2h

3h

4h

6h

2000 mg/kg bw

5638

Normal

+

+

+

+

+

+

+

+

21/21

5640

Normal

+

+

+

+

+

+

+

+

21/21

5646

Normal

+

+

+

-

+

+

+

+

20/21

Deceased activity

-

-

-

+

-

-

-

-

1/21

Hunched back

-

-

-

+

-

-

-

-

1/21

Piloerection

-

-

-

+

-

-

-

-

1/21

 

Group 2

Animal Number

Observations

Day 0

Day 1-14

F

15’

30’

1h

2h

3h

4h

6h

2000 mg/kg bw

5644

Normal

+

+

+

+

+

+

+

+

21/21

5645

Normal

+

+

+

+

+

+

+

+

21/21

5647

Normal

+

+

+

+

+

+

+

+

21/21

Comments: F = Frequency of observation = number of occurrence of observation / total number of observations per animal

              + = Observation present; - = Observation no present;

               ‘ = Minutes

              h = hour

 

NECROPSY FINDINGS

 

TEST ITEM: GELB SULFATO

TEST SYSTEM: CRL: (WI) BR RAT

STUDY CODE: 07/425-001P

MODE OF ADMINISTRATION: ORAL

SEX: FEMALE

DURATION OF STUDY: 14 DAYS

DOSE: 2000 MG/KG BW

NUMBER OF ANIMALS: 3/3

DATE OF NECROPSY: 22 MAY 2007

NECROPSY FINDINGS PER ORGANS

ANIMAL NUMBERS:

5638

5640

5646

/

/

NECROPSY FINIDINGS

Σ

%

Uterus

Hydrometra

+

-

-

/

/

1

33

No organs with macroscopic findings

-

+

+

/

/

2

66

 

TEST ITEM: GELB SULFATO

TEST SYSTEM: CRL: (WI) BR RAT

STUDY CODE: 07/425-001P

MODE OF ADMINISTRATION: ORAL

SEX: FEMALE

DURATION OF STUDY: 14 DAYS

DOSE: 2000 MG/KG BW

NUMBER OF ANIMALS: 3/3

DATE OF NECROPSY: 23 MAY 2007

NECROPSY FINDINGS PER ORGANS

ANIMAL NUMBERS:

5644

5645

5647

/

/

NECROPSY FINDINGS

Σ

%

LUNGS

Pinprick-sized haemorrhages

+

-

-

/

/

1

33

No organs with macroscopic findings

-

+

+

/

/

2

66

COMMENT: NO ALTERATION = -

                     ALTERATION PRESENT = +

                    NO DATA = /

                    GRADE OF ALTERATION 1 = SLIGHT /SMALL / FEW

                                                   2 = MODERATE / M.SIZE / M. NUMBER

                                                   3 = MARKED / MANY / LARGE

Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral LD50 value of the test item Gelb Sulfato proved to be greater than 2000 mg/kg body weight in female CRL:(WI) BR rats.
Executive summary:

An acute oral toxicity study was performed to assess the acute toxicity of test item Gelb Sulfato in rats. The results of the study allowed the test item to be ranked according to the classification systems, currently in use.

 

The acute toxic class method according to OECD 423 and Directive 2004/73/EC B.1. tris was performed with Gelb Sulfato. The study was performed in accordance with the Principles of Good Laboratory Practice (GLP) and reported with a GLP certificate.

 

Two groups of three female CRL:(WI) BR Wistar rats were treated by single oral gavage with Gelb Sulfato at dose levels of 2000 mg/kg bw in two independent experiments (Treatment group 1 and Treatment group 2). A concentration of 200 mg/ml prepared with distilled water corresponding to a treatment volume of 10 ml/kg bw was used at both steps.

 

Clinical observations were performed for all animals 15 and 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Food was made available again 3 hours after the treatment. Body weight was measured weekly. Gross necropsy was performed on all animals at termination of examination (day 14).

 

Results

Mortality:No mortality occurred after a single oral administration of Gelb Sulfato at 2000 mg/kg bw dose level in female CRL:(WI) BR rats.

 

Treatment group:

1

2

Dose level (mg/kg bw):

2000

2000

Number of animals treated

3

3

Mortality:

0/3

0/3

 

Clinical symptoms:Slight activity decrease, hunched back and piloerection were noted for one animal (1/6) two hours after the treatment. There were no test item effect on the physical condition and behaviour of the remaining animals. Yellow coloured urine and faces were noted on the day of the treatment and on the next day.

 

Body weight and body weight gain:The mean body weight and the body weight gain of animals were in the normal range during the two weeks observation period, similar to the expected values in untreated animals of the same age and strain.

 

Necropsy

No macroscopic alterations related to the toxic effect of Gelb Sulfato were found

 

Conclusion:

The acute oral LD50value of the test item Gelb Sulfato was greater than 2000 mg/kg body weight in female CRL:(WI) BR rats.

Gelb Sulfato does not meet the requirements of EU labelling regulations.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

An acute oral toxicity study was performed to assess the acute toxicity of test item Gelb Sulfato in rats. The acute toxic class method according to OECD 423 and Directive 2004/73/EC B.1. tris was performed with Gelb Sulfato. Two groups of three female CRL:(WI) BR Wistar rats were treated by single oral gavage with Gelb Sulfato at dose levels of 2000 mg/kg bw in two independent experiments (Treatment group 1 and Treatment group 2). A concentration of 200 mg/ml prepared with distilled water corresponding to a treatment volume of 10 ml/kg bw was used at both steps. Clinical observations were performed for all animals 15 and 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Food was made available again 3 hours after the treatment. Body weight was measured weekly. Gross necropsy was performed on all animals at termination of examination (day 14).

No mortality occurred after a single oral administration of Gelb Sulfato at 2000 mg/kg bw dose level in female CRL:(WI) BR rats. Slight activity decrease, hunched back and piloerection were noted for one animal (1/6) two hours after the treatment. There were no test item effect on the physical condition and behaviour of the remaining animals. Yellow coloured urine and faces were noted on the day of the treatment and on the next day. The mean body weight and the body weight gain of animals were in the normal range during the two weeks observation period, similar to the expected values in untreated animals of the same age and strain.

No macroscopic alterations related to the toxic effect of Gelb Sulfato were found

In conclusion, the acute oral LD50 value of the test item Gelb Sulfato was greater than 2000 mg/kg body weight in female CRL:(WI) BR rats.

Gelb Sulfato does not meet the requirements of EU labelling regulations.

The acute dermal toxicity of test item Gelb Sulfato was assessed in rats to provide information on health hazards likely to arise from 24-hour exposure by the dermal route. A limit test according to OECD 402 and B.3. 92/69/EEC was performed with Gelb Sulfato in CRL: (WI) BR rats. Five male and five female animals were treated with Gelb Sulfato for a single 24-hour dermal exposure in its original form at a dose level of 2000 mg/kg bw. Clinical examinations were made on the day of treatment 1 h and 5 h after the application of the test item, and once each day for 14 days thereafter. The body weights of animals were recorded on day 0 and weekly thereafter. A gross necropsy was performed in all animals at the end of observation period.

No mortality occurred during the entire study period. Gelb Sulfato caused yellow – red discolouration of the treated skin for 2 - 9 days after the patch removal.

There were no behavioural changes or general toxic signs. Behaviour and general state of animals were considered to be normal. The body weight development was not influenced by the single dermal treatment with the test item Gelb Sulfato. Test item related specific macroscopic alterations were not found.

Under the conditions of the present study, single 24-hour dermal administration of the test item, Gelb Sulfato did not cause mortality and toxic dermal alterations in male and female CRL:(WI)BR rats. The acute dermal LD50 value of the test item Gelb Sulfato was greater than 2000 mg/kg bw in male and female CRL:(WI) BR rats.

Gelb Sulfato does not meet the criteria for classification according to EU labelling regulations for classification and labelling of dangerous substances.

The test substance has a presumed very low vapour pressure and is a granular or well dedusted product, hence the potential for the generation of inhalable forms is low. In addition, production and use is done in a closed process without isolation of reaction products. The isolated product are dust free granules or well dedusted powder (non-dusty solid) which may be formulated into a liquid preparation of low volatility and the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur. Dermal exposure is considered to be the appropriate route of exposure and has been assessed accordingly. No acute inhalation test was performed.

Justification for classification or non-classification

The above studies have all been ranked reliability 1 according to the Klimisch et al system. This ranking was deemed appropriate because the studies were conducted to GLP and in compliance with agreed protocols. Sufficient dose ranges and numbers are detailed; hence it is appropriate for use based on reliability and animal welfare grounds.

The above results triggered no classification under the Dangerous Substance Directive (67/548/EEC) and the CLP Regulation (EC No 1272/2008). No classification for acute effects is therefore required.