JAUNE REACTIF 181

Endpoint summary

Description of key information

Repeated dose toxicity- oral:

A 5-day range finding study was performed to select the dosages for a 28-day oral toxicity study (RCC 1990). It was recommended to expose the animals to 0, 50, 200, or 1000 mg/kg bw of the test substance.

In a GLP-compliant repeated dose toxicity study, performed according to OECD guideline 407 and EU method B.7. Wistar rats were treated with the test substance (50, 200, and 1000 mg/kg bw) by repeated oral gavage, for a period of 28 days. The study was comprised of four groups, each containing five male and five female rats. For the post-exposure recovery period two satellite groups exposed to 0 and 1000 mg/kg bw for 28 days consisting of five male and five female rats each were monitored for an additional 14 days after the end of the 28 days treatment period. No treatment-related effects were observed on mortality, clinical signs, food consumption, body weight, ophthalmoscopic examinations, clinical laboratory investigations, macroscopic and microscopic findings. There was evidence that treatment of male rats with 1000 mg/kg of the test substance for 4 weeks was associated with a slight increase in liver weight which regressed during the treatment-free recovery period. There were no other differences in organ weight which could be attributed to treatment with the test substance. Based on the study results, the No Observed Adverse Effect Level of FAT 40400/A was determined to be 1000 mg/kg/bw.

 

Repeated dose toxicity- inhalation:

Currently no study to assess the repeated dose inhalation toxicity potential of Reactive Yellow 181 is available. However, the vapour pressure for the substance can be considered low owing to the high melting point (>300 °C). Hence, the substance is considered to have low volatility. Synthesis and formulation of this chemical is performed in a closed process; the final product consists of liquid formulations only. Hence, the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Further, Reactive Yellow 181 was found to be miscible in water (water solubility 332 g/L) and have low log partition coefficient (-10), hence, in the case of dust of the substance entering the respiratory tract, it will be trapped in the mucus and cleared, thereby further limiting the absorption. Further, no adverse effects were observed in a 28-day repeated dose oral toxicity study with Reactive Yellow 181, where the NOAEL was found to be 1000 mg/kg bw/day. Taking into consideration the above arguments, low toxicity potential is expected on repeat exposure of Reactive Yellow 181 via inhalation route and hence, repeated dose toxicity testing by the inhalation route was considered scientifically not necessary.

 

Repeated dose toxicity- dermal:

Currently, no study to assess the repeated dose dermal toxicity potential of Reactive Yellow 181 is available. However, the molecular weight of the chemical is 828.1 g/mol, indicating it being too large for dermal absorption. It has water solubility of 332 g/L and n-octanol/water partition coefficient (log P) of -10, indicating it being too hydrophilic to cross the lipid rich environment of the stratum corneum in the epidermis. Hence, the dermal uptake for the substance will be low. In addition, the use of this substance will not result in aerosols, particles or droplets, so exposure to humans via the dermal route will be unlikely to occur. The chemical showed low toxicity potential in the available acute oral (LD₅₀ >5000 mg/kg bw) and dermal (LD₅₀ >2000 mg/kg bw) toxicity studies. Further, no adverse effects were observed in a 28-day repeated dose oral toxicity study with Reactive Yellow 181, where the NOAEL was found to be 1000 mg/kg bw/day. Hence, safety for human health can be estimated via route to route extrapolation. Similarly, absence of systemic toxicity in eye and skin irritation as well as in skin sensitisation studies, further supports the conclusion that low toxicity is expected for the chemical via the dermal route. Taking  the above arguments into consideration, low toxicity potential is expected on repeated exposure of Reactive Yellow 181 via dermal route and hence, repeated dose toxicity testing by the dermal route was considered scientifically not necessary.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 July 1990 to 4 September 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

Test substance: FAT-40400/A
Expiry Date: 3/95
Stability of test article: Stable
Stability of test article in vehicle: stable for at least 2 hours
Storage: in the original container, at room temperature, protected from light

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wölferstrasse 4, CH-4414 Füllinsdorf
- Age at acclimation: 6 weeks
- Weight at acclimatization: males: 159 - 177 g, females: 161 - 175 g
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz)
- Diet: Pelleted standard Kliba 343, Batches 70/90, 71/90, 73/90 rat maintenance diet ("Kliba" Klingentalmuehle AG, CH-4303 Kaiseraugst), available ad libitum
- Water: Community water from Itingen, available ad libitum
- Acclimation period: Seven days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration, homogeneity and stability of the mixtures were determined during acclimatization. Further samples for analysis were taken during
acclimatization and during week 3 of the test, than kept deep frozen until later determination of analysis.

Duration of treatment / exposure:

up to 28 days

Frequency of treatment:

daily

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Remarks:

Control

Dose / conc.:

50 mg/kg bw/day (actual dose received)

Remarks:

Low dose

Dose / conc.:

200 mg/kg bw/day (actual dose received)

Remarks:

Middle dose

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Remarks:

High dose

No. of animals per sex per dose:

The number of rats assigned to toxicity and recovery testing per group were:
- 5 per sex for 50 and 200 mg/kg bw for the toxicity test
- 5 per sex for 0 and 1000 mg/kg bw for the recovery test

Control animals:

yes, concurrent vehicle

Details on study design:

FAT 40400/A was administered to SPF-bred Wistar rats by repeated oral gavage, for a period of 28 days. The study was comprised of four groups, each containing five male and five female rats. The test article/vehicle mixture was administered on a mg/kg bw basis by oral gavage. The dose was based upon data received from acute studies and a 5-days oral (range-finding) gavage study. The animals of the control group were treated similarly with the vehicle alone. For the post-exposure recovery period two satellite groups exposed to 0 and 1000 mg/kg bw for 28 days consisting of five male and five female rats each were monitored for an additional 14 days after the end of the 28-days treatment period.

Observations and examinations performed and frequency:

- Mortality/viability: Observations for mortality were recorded once daily.
- Clinical signs: Signs of toxicity were assessed once daily. Descriptions of all abnormalities were recorded and the subsequent progress was monitored.
- Food consumption: The food consumption was recorded once during the acclimatization period and weekly thereafter.
- Body weights: The body weight of each animal was recorded on the same days as the food consumption.
- Ophthalmoscopic examination: Ophthalmoscopic examinations were performed on all animals. A description of any abnormality was recorded. Examinations were performed at termination of treatment and a second time on the recovery individuals of groups 1 and 4 at termination of the recovery period. Ten minutes after the application of a mydriatic solution (Dispersa AG, CH-8400 Winterthur) the cornea, lens, anterior chamber, vitreous body and ocular fundus of both eyes were examined under dimmed light using a Heine Miroflex 2 Ophthalmoscope (Eisenhut Vet. AG, CH-4123 Allschwil).
- Haematology/clinical chemistry: Blood samples for hematology and clinical biochemistry were collected from all animals under light ether anesthesia at termination of treatment. The animals were fasted for approximately 18 hours before blood sampling but water was provided. Blood samples were collected from each animal between the hours of 7.00 and 9.30 a.m. to reduce biological variation caused by circadian rhythms. Blood samples were drawn from the retro-orbital plexus. Parameters being measured: erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet count, reticulocyte count, nucleated erythrocytes normoblasts, heinz bodies, methemoglobin, total leukocyte count, differential leukocyte count, red cell morphology, thromboplastin time, activated partial thromboplastin time, glucose, urea, creatinine, uric acid, bilirubin, cholesterol, triglycerides, phospholipids, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, creatine kinase, alkaline phosphatase, gamma-glutamyl-transferase, calcium, phosphorus, sodium, potassium, chloride, albumin, protein total.
- Urinalysis: Urine was collected during the 18-hour fasting period into a specimen vial using a metabolism cage, during which time the animals were deprived of food but allowed access to water ad libitum. Parameters being measured: volume (18 hour), specific gravity, pH, color, protein, glucose, ketone, bilirubin, blood, urobilinogen, urine sediment.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
- All animals were necropsied and descriptions of all macroscopic abnormalities were recorded. The following organ weights were taken from all animals necropsied at termination of treatment or recovery: adrenals, brain, heart, kidneys, liver, ovaries, pituitary gland, spleen, testes, thyroid gland.

HISTOPATHOLOGY: Yes
- Slides of adrenals, heart, kidneys, liver, spleen and stomach collected at terminal sacrifice from the animals of the control and high-dose groups were examined by a pathologist. The same applied to all gross lesions and to all animals which died spontaneously or had to be killed in extremis. Upon detection of treatment-related morphologic changes in the organs of any high-dose animal, histologic evaluation of the same organs in all dose groups were performed. All abnormalities were described and included in the report.

Statistics:

The following statistical methods were used to analyze the body weights, food consumption, organ weights and clinical laboratory data: Univariate one-way analysis of variance was used to assess the significance of intergroup differences. If the variables could be assumed to follow a normal distribution, the Dunnett-test (many to one t-test) based on a pooled variance estimate was applied for the comparison between the treated groups and the control groups for each sex. The Steel-test (many-one rank test) was applied when the data could not be assumed to follow a normal distribution.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

The lower values seen in males receiving 50 mg/kg FAT 40400/A are considered not to be due to treatment because of the lack of a dose-relationship.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

The lower values seen in males receiving 50 mg/kg FAT 40'400/A are considered not to be due to treatment because of the lack of a dose-relationship.

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

However, in the urinalysis 5 out of 10 male rats of group 4 (1000 mg/kg) indicated a deep yellow urine discoloration at termination of the treatment. This observation was attributed to the high concentration and yellowish nature of the test article and is not an indication of renal insufficiency. At termination of the treatment-free recovery period this finding was no longer observed.

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

The following conclusions are based on examination of absolute organ weights, organ/body weight ratios and organ/brain weight ratios. In males, although there were no statistically significant differences in absolute organ weight between treated and control group means, there was evidence from organ/body and organ/brain weight ratios of a slight increase in liver and heart weights at 1000 mg/kg FAT 40400/A after 4 weeks of treatment. After the 14-day treatment-free recovery period there was no evidence of a treatment-related effect. In females, heart weight in rats receiving FAT 40'400/A was lower than in controls after 4 weeks of treatment but the fact that the differences were greater in the groups receiving 50 or 200 mg/kg than in the high dose group receiving 1000 mg/kg FAT 40400/A suggests that this was not due to treatment. Similarly, although liver weight in high dose females after the 14-day treatment-free recovery period was higher than in control females, the absence of any treatment-related difference at the end of the dosing period suggest that this was not due to the administration of FAT 40400/A.

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Effect level based on clinical symptoms, body weight, food consumption, ophthalmoscopic examination, clinical laboratory investigations, and pathology

Critical effects observed:

no

Conclusions:

The No Observed Adverse Effect Level (NOAEL) of FAT 40400/A was determined to be 1000 mg/kg/bw.

Executive summary:

In a GLP-compliant repeated dose toxicity study, performed according to OECD guideline 407 and EU method B.7. Wistar rats were treated with the test substance (50, 200, and 1000 mg/kg bw) by repeated oral gavage, for a period of 28 days. The study was comprised of four groups, each containing five male and five female rats. For the post-exposure recovery period two satellite groups exposed to 0 and 1000 mg/kg bw for 28 days consisting of five male and five female rats each were monitored for an additional 14 days after the end of the 28-days treatment period. No treatment-related effects were observed on mortality, clinical signs, food consumption, body weight, ophthalmoscopic examinations, clinical laboratory investigations, macroscopic and microscopic findings. The lower values of food consumption seen in males receiving 50 mg/kg FAT 40400/A are considered not to be due to treatment because of the lack of a dose-relationship. However, in the urinalysis 5 out of 10 male rats of group 4 (1000 mg/kg) indicated a deep yellow urine discoloration at termination of the treatment. This observation was attributed to the high concentration and yellowish nature of the test article and is not an indication of renal insufficiency. At termination of the treatment-free recovery period this finding was no longer observed. There was evidence that treatment of male rats with 1000 mg/kg of the test substance for 4 weeks was associated with a slight increase in liver weight which regressed during the treatment-free recovery period. There were no other differences in organ weight which could be attributed to treatment with the test substance. So, based on the study results, the No Observed Adverse Effect Level (NOAEL) of FAT 40400/A was determined to be 1000 mg/kg/bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

GLP compliant guideline study, klimisch 1

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the findings of the repeated dose toxicity study, the test substance does not need to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.