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Administrative data

Description of key information

No classification for acute effects is required.

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:
09 August to 24 August 2011
Reliability:
1 (reliable without restriction)
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:
During the animal exposures, the animal room temperature was in the range 24.3 – 26.2 °C,from 16 August 2011 to 17 August 2011. This was a deviation from the range stated in the guidelines (19.0 – 25.0 °C).
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
During the animal exposures, the animal room temperature was in the range 24.3 – 26.2 °C,from 16 August 2011 to 17 August 2011. This was a deviation from the range stated in the guidelines (19.0 – 25.0 °C).
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
During the animal exposures, the animal room temperature was in the range 24.3 – 26.2 °C,from 16 August 2011 to 17 August 2011. This was a deviation from the range stated in the guidelines (19.0 – 25.0 °C).
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
EXPERIMENTAL ANIMALS

Species and strain: RjHan:WI rats
Source: Laboratoire Elevage Janvier, B.P. 4105, Route des Chênes Secs, 53940 Le Genest-St-Isle CEDEX FRANCE
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals, 3 animals/group
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, 10 weeks old
Date of receipt: 28 July 2011
Body weight at treatment: 221 – 230 g
Acclimation period: At least 12 days

Husbandry

Animal health: Only healthy animals were used for the test. The veterinarian certified the health status.
Number of animal room: 522/10
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: Lignocel Bedding for Laboratory Animals was available to animals during the study. A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 21.4 – 26.2°C
Relative humidity: 30 - 70%
Ventilation: 15-20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

The temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply

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 ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Animal Identification

Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File, for each animal allocated to the treatment groups. 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: Distilled water
Dispense Code: 0110111
Expiry Date: 31 January 2014
Dose volume: 10 mL/kg bw

Test item was freshly formulated at a concentration of 200 mg/mL in the vehicle (without correction for purity), in the Central Dispensary Unit of CiToxLAB Hungary Ltd. on the day of administration. The formulation container was stirred continuously during administration to ensure that the syringe was filled from a homogeneous suspension/solution.

Justification of the dose:

The initial dose level was selected on the basis of the information provided by the Sponsor. The LD50 value was expected to be above 2000 mg/kg bw.

Initially, three female animals were treated with 2000 mg/kg bw of Reactive Yellow F01-0555. As no mortality occurred within 24 hours after dosing, a second group of three animals received 2000 mg/kg bw of the test item approximately 24 hours after treatment of the first group. No mortality occurred in the second treatment group; hence, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris). The test was terminated on completion of the 14-day observation period.

Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Initially, three female animals were treated with 2000 mg/kg bw of Reactive Yellow F01-0555. As no mortality occurred within 24 hours after dosing, a second group of three animals received 2000 mg/kg bw of the test item approximately 24 hours after treatment of the first group.
Control animals:
no
Details on study design:
Procedure

A single oral dose was administered by gavage followed by a fourteen-day observation period. The animals were fasted for about 16 hours prior to treatment. The food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment. A constant treatment volume of 10 mL/kg bodyweight was applied.


OBSERVATIONS

Clinical Observations

Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily 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. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement

The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0), Day 3, Day 7 and Day 14.

NECROPSY

Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthasol® 40 %; details are presented in 3.1.3.). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

EVALUATION OF THE RESULTS

The method used was not intended to allow the calculation of a precise LD50 value.
The test item was ranked into categories of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423.
Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.

Statistics:
None
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Reactive Yellow F01-0555 did not cause mortality at a dose level of 2000 mg/kg bw.
Clinical signs:
other: Treatment with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw caused orange urine and faeces on Day 1 in all Group 1 animals and at 6 hours after treatment in all Group 2 animals. All animals fully recovered and were free of any clinical signs
Gross pathology:
There were no test item related macroscopic findings in animals dosed at 2000 mg/kg bw at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the test item Reactive Yellow F01-0555 was found to be above 2000 mg/kg bw in female RjHan:WI rats.
Executive summary:

A single-dose oral toxicity of Reactive Yellow F01-0555 was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris) in RjHan:WI rats. Two groups of three female RjHan Wistar rats (10 weeks old) were treated with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw (Group 1 and Group 2).

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. Rats were maintained without compound administration for a 2‑week observation period after the day of dosing. As no mortality was observed in this dose group within 24 hours after dosing, a confirmatory treatment was performed on 3 further females (Group 2) at the same dose level. As no mortality was observed in the second dose group, no further testing was required according to OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris.

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal (about 16 hours prior to treatment). Food was made available again 3 hours after the treatment. Reactive Yellow F01-0555 was administered formulated in distilled water at a concentration of 200 mg/mL at a dosing volume of 10 mL/kg bw. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 3 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

Reactive Yellow F01-0555 did not cause mortality at a dose level of 2000 mg/kg bw. Treatment with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw caused orange urine and faeces in all animals (6/6). All animals fully recovered and were freeof any clinical signsfrom Day 2 until the end of the observation period. Body weights or bBody weight gains showed no indication of a treatment-related effect.

There were no test item related in animals dosed at 2000 mg/kg bw at necropsy.

Under the conditions of this study, the acute oral LD50 value of the test item Reactive Yellow F01-0555 was found to be above 2000 mg/kg bw in female RjHan:WI rats.

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
GLP study conducted in accordance with OECD, EU and US guidelines therefore study categorised as Klimisch 1 study and assessment factor for the quality of whole database is 1.

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:
24 August 2011 to 07 September 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
On occasion during the study, the temperature (22 ± 3°C ) was recorded out of the target range. The actual range was at the temperature 20.7- 26.2 °C.
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
EXPERIMENTAL ANIMALS
Species and strain: CRL:(WI)BR Wistar rats
Source: CHARLES RIVER (EUROPE) LABORATORIES INC.
Hygienic level at arrival: SPF
Hygienic level during
the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies
Number of animals: 5 animals/sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at study start: Young adult rats
Body weight range
at dosing: Between 215 g and 280 g
Acclimatization time: 13 days

Husbandry
Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room-Box: 242/5
Housing: Individual caging
Cage type: Type II. polypropylene/polycarbonate
Bedding: Laboratory bedding:
Lignocel Hygienic Animal Bedding produced by J. Rettenmaier & Söhne GmbH+Co.KG (Holzmühle 1, 73494 Rosenberg, Germany);
A copy of the relevant Certificate of Analysis is maintained in CiToxLAB Hungary Ltd.'s archive.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20.7- 26.2 °C
Relative humidity: 44 - 70 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
The temperature and relative humidity was recorded twice daily during the study.

Food and Water Supply
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 ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is not considered to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
The supplier provided an analytical certificate for the batch used.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Identification
The individual identification was performed using numbers written on the tail with a marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File for each animal allocated to the treatment groups. The cages were identified by cards containing information about study code, sex, dose group, cage number and individual animal numbers.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment. The test item was applied as a single dose as supplied to the shaved skin and remained in contact with the skin for the 24- hour exposure period. For that purpose, the appropriate amount of the test item was moistened with body temperature water and distributed as uniformly as possible. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.
At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
Justification of the dose: The test item was not expected to be lethal at 2000 mg/kg bw. A limit test was therefore performed.
No. of animals per sex per dose:
5 male + 5 female
Control animals:
no
Details on study design:
Clinical Observations
Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Measurement of Body Weight
The body weights were recorded on Day 0 (before test item administration) and on Days 3, 7 and 14.

NECROPSY
All animals were anaesthetised with Euthasol®40% (details in 3.1.3.) and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.
Statistics:
None
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:
other: No clinical signs were observed after the treatment with the test item or during the 14-day observation period.
Gross pathology:
There was no evidence of test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Pelvic dilation of the right kidney was seen as an incidental finding in one male rat (7931).
Other findings:
No local dermal signs were observed during the entire study period. However, orange staining was observed on the skin in all animals after dosing from Day 1 to Day 8.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item Reactive Yellow F01-0555 was found to be higher than 2000 mg/kg bw in male and female CRL:(WI)BR Wistar rats.
Executive summary:

An acute dermal toxicity study was performed with test item Reactive Yellow F01-0555 in CRL:(WI)BR Wistar rats, in compliance with OECD Guideline No.: 402. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied, moistened with distilled water, as a single dermal 24-hour exposure followed by a 14-day observation period. Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 3, 7 and 14. Rats were euthanized and a gross macroscopic examination performed at the end of the 2-week observation period (Day 14).

The results of the study were summarized as follows:

Mortality: No mortality occurred.

Systemic clinical signs: No clinical signs were observed after the treatment with the test item or during the 14-day observation period. Local dermal signs: No local dermal signs were observed during the entire study period. However, orange staining was observed on the skin in all animals after dosing from Day 1 to Day 8.

Body weight: One female showed slight bodyweight loss during the first week of the observation period.

Necropsy: There was no evidence of test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Pelvic dilation of the right kidney was seen as an incidental finding in one male rat (7931).

The acute dermal median lethal dose (LD50) of the test item Reactive Yellow F01-0555 was found to be higher than 2000 mg/kg bw in male and female CRL:(WI)BR Wistar rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
GLP study conducted in accordance with OECD, EU and US guidelines therefore study categorised as Klimisch 1 study and assessment factor for the quality of whole database is 1.

Additional information

A single-dose oral toxicity of Reactive Yellow F01-0555 was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris) in RjHan:WI rats. Two groups of three female RjHan Wistar rats (10 weeks old) were treated with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw (Group 1 and Group 2).

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. Rats were maintained without compound administration for a 2‑week observation period after the day of dosing. As no mortality was observed in this dose group within 24 hours after dosing, a confirmatory treatment was performed on 3 further females (Group 2) at the same dose level. As no mortality was observed in the second dose group, no further testing was required according to OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris.

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal (about 16 hours prior to treatment). Food was made available again 3 hours after the treatment. Reactive Yellow F01-0555 was administered formulated in distilled water at a concentration of 200 mg/mL at a dosing volume of 10 mL/kg bw. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 3 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

Reactive Yellow F01-0555 did not cause mortality at a dose level of 2000 mg/kg bw. Treatment with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw caused orange urine and faeces in all animals (6/6). All animals fully recovered and were freeof any clinical signsfrom Day 2 until the end of the observation period. Body weights or bBody weight gains showed no indication of a treatment-related effect.

There were no test item related in animals dosed at 2000 mg/kg bw at necropsy.

Under the conditions of this study, the acute oral LD50 value of the test item Reactive Yellow F01-0555 was found to be above 2000 mg/kg bw in female RjHan:WI rats.

 

An acute dermal toxicity study was performed with test item Reactive Yellow F01-in CRL:(WI)BR Wistar rats, in compliance with OECD Guideline No.: 402. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied, moistened with distilled water, as a single dermal 24-hour exposure followed by a 14‑day observation period. Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 3, 7 and 14.Ratswere euthanized and a gross macroscopic examination performed at the end of the 2-week observation period (Day 14).

During the study, no mortality was observed. Furthermore, no clinical signs were observed after the treatment with the test item or during the 14‑day observation period. In addition, no local dermal signs were observed during the entire study period. However, orange staining was observed on the skin in all animals after dosing from Day 1 to Day 8.

One animal female showed slight bodyweight loss during the first week of the observation period.

There was no evidence of test item-related observations at a dose level of 2000 mg/kg bw at necropsy.

The acute dermal median lethal dose (LD50) of the test itemReactive Yellow F01‑0555was found to be higher than 2000 mg/kg bw in male and femaleCRL:(WI)BR Wistar rats.

Acute toxicity: Inhalation.

The test substance has a predicted very low vapour pressure and is a granular product, hence the potential for the generation of inhalable forms is low. In addition 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.