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Administrative data

Description of key information

Key value for chemical safety assessment

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:
Delivery of Animals: December 02, 1996 - Termination: January 20, 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: Directive 92/69/EEC, B. 7. "Subacute Toxicity Oral"
Version / remarks:
July 31, 1992
Qualifier:
according to guideline
Guideline:
other: "Japanese Guidelines for Screening Toxicity Testing of Chemicals: Testing Methods for New Chemical Substances"
Version / remarks:
enacted July 13, 1974, amended December 5,1986
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Hanlbm:WIST (SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4 4414 Fuellinsdorf/Switzerland
- Age at study initiation (at delivery): 6 weeks
- Weight at study initiation: At acclimatization: Males: 127-176 grams (mean 150 grams); Females: 98-130 grams (mean 114 grams)
- Housing: Groups of five in Makrolon type-4 cages with standard softwood bedding ('Lignocel' Schill AG , 4132 Muttenz /Switzerland).
- Diet (e.g. ad libitum): Pelleted standard Kl i ba no. 343 rat maintenance di et (KLIBA MUhl en AG, 4303 Kaiseraugst/Switzerland) was available ad libitum (Batch no. 81/96) . The feed batch was analyzed for contaminants.
- Water (e.g. ad libitum): Tap water was available ad libitum in water bottles. Results of bacteriological assays, chemical and contaminant analysis of representative samples are attached to this report
- Acclimatization: 7 days under test conditions following a health examination. Only animals without any visible signs of illness were used for the study.

DETAILS OF FOOD AND WATER QUALITY: None of the contaminants analysed in the water and diet is considered to have been present at a concentration which would have affected the validity of the results.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12, (light period between 6.00 a.m. and 6.00 p.m.), music during the light period.

IN-LIFE DATES:
Allocation A (toxicity testing: termination after 28 treatment days) :
Allocation B (recovery testing: termination after an additional 14-day recovery period):
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
bi-distilled water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:

Frequency of preparation: daily, prior to each application
SCARLET RN 1165 was weighed into a glass beaker on a tared Mettler balance and the vehicle added. The mixtures were prepared using a magnetic stirrer.
Homogeneity of the test article in the vehicle was maintained during treatment using a magnetic stirrer.

VEHICLE
bi-distilled water
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentration, homogeneity and stability of the test article/vehicle mixtures were determined in samples taken during acclimatization and during week 3 of the treatment. The analyses were performed in the Analytical Laboratories of RCC Umweltchemie AG according to a method supplied by the sponsor (for results please refer to Attachment 4, pp. 219-233 of the study report).

Chemical analysis of dose preparations: at acclimatization 04-DEC-1996, at 3 weeks 24-DEC-1996
Duration of treatment / exposure:
Duration of acclimatization : 7 days
Duration of treatment : 28 days
Duration of recovery : 14 days
Frequency of treatment:
Once daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5 for Allocation A - toxicity testing: termination after 28 treatment days
5 additional only for 0 (control) and 1000 mg/kg bw/day groups for Allocation B - recovery testing: termination after an additional 14-day recovery period
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based upon the results of a non-GLP 5-day dose- range-finding study (RCC Study Number 816028) in which BLUE GS 5664.80 was administered by gavage to 2 rats per group and sex.
- Rationale for dose level selection: Based upon data from an acute toxicity study (RCC Project 639685) and a 5-day dose-range-finding study (RCC Project 639742) in which SCARLET RN 1165 was administered by gavage to 3 rats per sex and dose group.
Between test days 3 to 5 the absolute and relative food consumption was statistically significantly higher in males treated at 1000 mg/kg when compared with the controls. As the food consumption of these animals was already higher during the pretest, no toxicological relevance was attributed to this findings.
- Post-exposure recovery period in satellite groups: 14 days
Positive control:
None
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: body weights were recorded weekly during pretest, weeks 1-3 and 5. Twice weekly in weeks 4 and 6.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Time schedule for examinations: The food consumption was recorded once during the pretest period and weekly thereafter using an on-line electronic recording system consisting of a Mettler balance connected to the RCC computer
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not specified

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at pretest 06-DEC-1996, at 4 weeks 03-JAN-1997 and at 6 weeks 17-JAN-1997
- Dose groups that were examined: Ophthalmologic findings were noted in a small proportion of animals from all groups. They included persistent pupillary membranes and corneal opacity. These findings occurred at similar incidences in the control and treated groups at the end of the treatment period. Therefore, they are considered to be unrelated to treatment with test article.

HEMATOLOGY: Yes
- Time schedule for collection of blood: after 4 weeks 06-JAN-1997 and after 6 weeks 20-JAN-1997, Blood samples were collected between the hours of 06.15 and 07.50 to reduce biological variation caused by circadian rhythms
- Anaesthetic used for blood collection: Yes- Blood samples for hematology and clinical biochemistry were collected from all animals under light ether anesthesia.
- Animals fasted: Yes -The animals were fasted in metabolic cages for approximately 18 hours before blood sampling but allowed access to water ad libitum
- How many animals: all animals
- Parameters checked: Erythrocyte count; Hemoglobin; Hematocrit; Mean corpuscular volume; Mean corpuscular hemoglobin; Mean corpuscular hemoglobin concentration; Platelet count; Reticulocyte count; Reticulocyte fluorescence ratios; Nucleated erythrocytes (normoblasts); Heinz bodies; Methemoglobin; Total leukocyte count; Differential leukocyte count; Red blood cell morphology; Thromboplastin time (=prothrombin time); Activated partial thromboplastin time. Please refer to pg. 20-21-22 of the study report for all parameters examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 4 weeks 06-JAN-1997 and after 6 weeks 20-JAN-1997, Blood samples were collected between the hours of 06.15 and 07.50 to reduce biological variation caused by circadian rhythms
- Animals fasted: Yes -The animals were fasted in metabolic cages for approximately 18 hours before blood sampling but allowed access to water ad libitum
- How many animals: all animals
- Parameters checked: Glucose; Urea; Creatinine; Uric acid; Bilirubin, total; Cholesterol, total; Triglycerides; Phospholipids; Aspartate aminotransferase; Alanine aminotransferase; Lactate dehydrogenase
Creatine kinase; Alkaline phosphatase; Gamma-glutamyl transferase; Calcium; Phosphorus; Sodium; Potassium; Chloride; Albumin; Protein, total; Globulin; Albumin/Globulin ratio. Please refer to pg. 23-26 of the study report for all parameters examined.

URINALYSIS: Yes
- Time schedule for collection of urine: Urine was collected during the 18-hour fasting period into a specimen vital. Blood sampling : after 4 weeks 06-JAN-1997 and after 6 weeks 20-JAN-1997
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Yes -The animals were fasted in metabolic cages for approximately 18 hours before blood sampling but allowed access to water ad libitum
- Parameters checked: Volume (18-hour); Specific gravity; Osmolality; Color; Appearance; pH; Protein; Glucose; Ketone; Bilirubin; Blood; Nitrite; Urobilinogen; Urine sediment. Please refer to pg. 27-29 of the study report for all parameters examined.

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER:
MORTALITY/VIABILITY: twice daily
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- All animals
- Necropsy after 4 weeks 06-JAN-1997; after 6 weeks 0-JAN-1997
- Tissus and organs collected:
Adrenal glands, aorta, bone - sternum and femur; bone marrow - femoral and sternal; brain, epididymides, esophagus, eyes with optic nerve and Harderian gland, heart, joint - femorotibial, kidneys, large intestine - cecum, colon and rectum; lacrimal glands - exorbital, larynx, liver, lungs, lymph nodes - mesenteric and mandibular; mammary gland area, nasal cavity, ovaries, pancreas, pituitary gland, prostate gland, salivary glands - mandibular and sublingual; sciatic nerve, seminal vesicles, skeletal muscle, skin, small intestine - duodenum, jejunum and ileum; spinal cord - cervical, midthoracic and lumbar segments; spleen, stomach, testes, thymus, thyroid glands with parathyroid glands, tongue, trachea, urinary bladder, uterus, vagina and all gross lesions.

ABSOLUTE AND RELATIVE ORGAN WEIGHTS: Yes
The following organ weights were recorded on the scheduled dates of necropsy: Adrenals, brain, heart, kidneys, liver, ovaries, pituitary, spleen, testes, thyroid including parathyroid gland.
The organ to terminal body weight ratios as well as the organ to brain weight ratios were determined.
The determination of the terminal body weight was performed immediately prior to necropsy.

HISTOPATHOLOGY: Yes
Slides of adrenals, heart, kidneys, liver, lungs, spleen, stomach and testes collected at terminal sacrifice from the animals of group 1 (control) and group 4 (high-dose) as well as the stomach and gross lesions from rats of all groups were examined by a pathologist.
Other examinations:
DATA CALCULATION

FOOD CONSUMPTION
The food consumption was calculated per rat and per food consumption interval. It expresses the average food consumed per animal and per day over the food consumption interval.
FC = C/AD
where
FC is Food consumption in grams of food per animal and day;
C is measured food consumption in grams per cage over the consumption interval and AD is total consumption days over all animals in the cage during the consumption interval (the mortality during the consumption interval is therefore taken into account).

RELATIVE FOOD CONSUMPTION
The relative food consumption was calculated according to the following formula:
RFC = [FC/BW(i)]x1000
where
BW(i) is the most ideal body weight in grams or the body weight (of the corresponding rats) recorded on the day most close to the middle of the food consumption interval. In cases of equal "closeness" of two body weight records the later one was chosen;
RFC is relative food consumption in grams of food per kg body weight and day, and FC is food consumption in grams of food per animal and day (gram).
Unit is gram food per kg body weight and day
Statistics:
The following statistical methods were used to analyze the body weights, organ weights all ratios as well as clinical laboratory data :
When 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.
The Fisher's exact test was applied to the ophthalmoscopy data and macroscopical findings.

References :
- C.W. Dunnett: A Multiple Comparison Procedure for Comparing Several Treatments with a Control, J. Amer. Stat. Assoc. 50, 1096-1121 (1955).
- R.G. Miller: Simultaneous Statistical Inference, Springer Verlag, New York (1981).
- R.A. Fisher: Statistical Methods for Research Workers, Oliver and Boyd, Edinburgh (1950).
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
There were no clinical signs in group 2 (50 mg/kg) and in females of group 3 (200 mg/kg). Red feces were noted in males of group 3 (200 mg/kg). In all animals of group 4 (1000 mg/kg) red urine and red feces were recorded. The discoloration proved to be reversible during the recovery period, allowed to rats of group 4 (1000 mg/kg).
Mortality:
no mortality observed
Description (incidence):
All animals survived their assigned study period.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No differences from controls of statistical or biological significance were noted.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There was no effect on food consumption.
Slightly lower food consumption was recorded in males of group 3 (200 mg/kg), only. In the absence of any effect on food consumption in rats of group 4 (1000 mg/kg), this difference from the control is considered to be incidental.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Ophthalmologic findings were noted in a small proportion of animals from all groups. They included persistent pupillary membranes and corneal opacity. These findings occurred at similar incidences in the control and treated groups at the end of the treatment period. Therefore, they are considered to be unrelated to treatment with test article.
Haematological findings:
no effects observed
Description (incidence and severity):
Treatment period and Recovery period - no remarkable findings were noted.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Treatment period - Slightly decreased glucose level in females of group 4 (p<0.01), marginally increased creatinine level in males of group 4 (p<0.05), slightly increased uric acid level in both sexes of group 4 (p<0.01), slightly to markedly increased total bilirubin level in both sexes of groups 3 and 4 (p<0.05 - p<0.01), slightly increased triglyceride level in both sexes of group 4 (p<0.01), slightly decreased calcium level (p<0.01) and marginally increased chloride level (p<0.05) in females of group 4.
In addition, plasma discoloration was observed in all animals of groups 3 and 4, ranging from a light red in group 3 to red in group 4.

Recovery period - all findings recorded during the treatment period were found to be reversible and generally similar to those of the controls. However, a slightly but statistically significantly increased uric acid and total bilirubin level in males (p<0.05 - p<0.01), and slightly increased chloride level in females (p<0.05) were yet to be noted. Subsequently, no further plasma discoloration was observed.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
Treatment period- Apart from urine discoloration, no remarkable findings were noted. Urine discoloration as observed in both sexes of groups 3 and 4 ranged from a deep-yellow (2 females) to light-orange (4 males) to orange (1 male) in
group 3 to orange (1 male, 8 females) to deep-orange (2 males, 1 female) to red (7 males, 1 female) in group 4. Moreover, a deep-yellow urine discoloration was also noted in 3 out of 5 males and 2 out of 5 females of group 2 ( 50 mg/kg).
Recovery period - Reversibility of the urine discoloration was observed, however, a deep-yellow discoloration was yet to be noted in all males and in two out of five females of the recovery group .
Due to the extreme test article-related discoloration of the plasma samples in groups 3 and 4, the biological relevance of the clinical biochemistry findings to the treatment remains doubtful. All other statistical differences in the results of the hematology, cltnical biochemistry or urinalysis parameters during the treatment or recovery period were considered to be incidental and unrelated to the treatment, and of normal biological variation for rats of this strain and age (for reference, refer to historical control data for untreated Wistar HanIbm rats, pp. 200-217 of the study report)
Behaviour (functional findings):
not specified
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
The absolute and relative organ weights did not distinguish treated groups from controls.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Reddish discoloration was noted mainly in the disgestive tract and kidneys from primarily rats of group 4 and 3 at the end of the main study period. This was due to the test article, a dyestuff and was not accompanied by microscopic
alterations .
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Miscroscopic findings:
In rats of both sexes from group 4 at the end of the main study period, there was a slight increase in the incidence and severity compared to the controls, of minimal to slight degrees of apoptotic bodies in the parietal cells of the gastric mucosa. This alteration was accompanied by minimal degree of vacuolation of the squamous epithelium of the limiting ridge and a slight increase in the incidence and severity of minimal to slight inflammatory cell infiltration of the glandular submucosa.
These findings were also present to a lesser degree in group 3. At the end of the recovery period these findings had largely reversed to background levels .
The remainder of microscopic findings recorded were within the normal range of background morphologic alterations which may be recorded in this strain and age of Wistar rat and occurred at similar incidences and severity in both control and treated rats at both sacrifices.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
DISCUSSION OF THE RESULTS
The oral administration of SCARLET RN 1165 to Wistar rats at doses of 50, 200 and 1000 mg/kg/day, for 28 days, resulted in no effects on mortality, food consumption, body weight, ophthalmoscopic examinations and organ weights.

Treatment with test article , a pigment, resulted in red feces in males at 200 mg/kg. Red feces and red urine was observed in all animals at 1000 mg/kg.

Test article-related changes of clinical biochemistry and urinalysis comprised the following parameters: slightly decreased glucose level in females of group 4 (p<0.01), marginally increased creatinine level in males of group 4 (p<0.05), slightly increased uric acid level in both sexes of group 4 (p<0.01), slightly to markedly increased total bil i rubin level in both sexes of groups 3 and 4 (p<0.05 - p<0.01), slightly increased triglyceride level in both sexes of group 4 (p<0.01), slightly decreased calcium level (p<0.01) and marginally increased chloride level (p<0.05) in females of group 4.

Plasma discoloration and urine discoloration was observed in all animals of groups 3 and 4. Urine discoloration was also present in some animals of group 2 (50 mg/kg).

Recovery period - all of the findings recorded during the treatment period were found to be reversible and generally similar to those of the controls. However, for clinical biochemistry data a slightly, but statistically significantly increased uric acid and total bilirubin level in males (p<0.05 - p<0.01), and slightly increased chloride level in females (p<0.05) were yet to be noted.
Subsequently, no further plasma discoloration was observed, whereas urinalysis indicated a deep-yellow urine discoloration in all males and in two out of five females.
Reddish discoloration was noted mainly in the digestive tract and kidneys from primarily rats of group 4 and 3 at the end of the main study period. This was due to the test article, a dyestuff and was not accompanied by microscopic
alterations.

Morphologic alterations following the administration of the test article were present in the stomach. These took the form of minor degrees of apoptotic bodies in parietal cells accompanied by also minor degrees of vacuolation of limiting ridge epithelium and inflammatory cell infiltration of the glandular submucosa. These findings had largely reversed following the recovery period. Both sexes of group 4 (1000 mg/kg) were clearly affected, group 3 (200 mg/kg) to a lesser degree. These findings may be regarded as indicators of a slight irritant effect of the test article on the gastric mucosa.

Based on the results of this study, 50 mg/kg SCARLET RN 1165 was established as the no-observed-adverse-effect level (NOAEL). Under the conditions of this study a no-observed-effect-level (NOEL) could not be determined.
Key result
Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: non-neoplastic
mortality
ophthalmological examination
organ weights and organ / body weight ratios
urinalysis
Key result
Critical effects observed:
no
Conclusions:
Oral administration of SCARLET RN 1165 to Wistar rats at doses of 0, 50, 200 and 1000 mg/kg/day, performed according to OECD 407/method EC B7, for 28 days resulted in the following:
Based on the results of this study, 50 mg/kg SCARLET RN 1165 was established as the no-observed-adverse-effect level (NOAEL).
Executive summary:

GENERAL

In this subacute toxicity study, SCARLET RN 1165 was administered daily by oral gavage to SPF-bred Wistar rats of both sexes at dose levels of 0, 50, 200 and 1000 mg/kg body weight/day for a period of 28 days.

The study comprised 5 animals per group and sex which were sacrificed after 28 days of treatment. Additional 5 rats per sex and group were used at 0 and 1000 mg/kg. These animals were treated for 28 days and then allowed a 14-day treatment-free recovery period after which they were sacrificed.

Clinical signs, food consumption and body weights were recorded periodically during the acclimatization, treatment and recovery period. Ophthalmoscopic examinations were performed during acclimatization, at the end of the treatment and recovery period.

At the end of the dosing period and the treatment-free period, blood samples were withdrawn for hematology and plasma chemistry analyses, and urine samples were collected for urinalysis. All animals were killed, necropsied and examined post mortem. Samples of major organs from all group 1 (0 mg/kg) and group 4 (1000 mg/kg) animals, as well as stomach and gross lesions from all animals were processed as hematoxylin and eosin stained slides and examined by light microscopy.

The results of the study are summarized as follows:

MORTALITY

All animals survived their assigned study period.

CLINICAL SIGNS

There were no clinical signs in group 2 (50 mg/kg) and in females of group 3 (200 mg/kg). Red feces were noted in males of group 3 (200 mg/kg). In all animals of group 4 (1000 mg/kg) red urine and red feces were recorded. The discoloration proved to be reversible during the recovery period, allowed to rats of group 4 (1000 mg/kg).

BODY WEIGHT

There was no effect on body weight.

FOOD CONSUMPTION/RELATIVE FOOD CONSUMPTION

There were no effects on food consumption or relative food consumption.

OPHTHALMOSCOPIC EXAMINATIONS

There were no treatment-related changes on the ophthalmological findings.

HEMATOLOGY/CLINICAL BIOCHEMISTRY/ URINALYSIS

In the assessment of hematology, clinical biochemistry and urinalysis data the following changes recorded between the control and treated rats of groups 3 (200 mg/kg) and/or 4 (1000 mg/kg) at termination of the treatment were considered to be test article related:

Hematology - no remarkable findings were noted.

Clinical Biochemistry - slightly decreased glucose level in females of group 4 (p<0.01), marginally increased creatinine level in males of group 4 (p<0.05), slightly increased uric acid level in both sexes of group 4 (p<0.01), slightly to markedly increased total bilirubin level in both sexes of groups 3 and 4 (p<0.05 - p<0.01), slightly increased triglyceride level in both sexes of group 4 (p<0.01), slightly decreased calcium level (p<0.01) and marginally increased chloride level (p<0.05) in females of group 4.

In addition, plasma discoloration was observed in all animals of groups 3 and 4, ranging from a light red in group 3 to red in group 4.

Urinalysis - except for urine discoloration, no remarkable findings were noted.

Urine discoloration as observed in both sexes of groups 3 and 4 ranged from a deep-yellow (2 females) to light-orange (4 males) to orange (1 male) in group 3 to orange (1 male, 8 females) to deep-orange (2 males, 1 female) to red (7 males, 1 female) in group 4. Moreover, a deep-yellow urine discoloration was also noted in 3 out of 5 males and 2 out of 5 females of group 2 ( 50 mg/kg).

Recovery period - all of the findings recorded during the treatment period were found to be reversible and generally similar to those of the controls. However, for clinical biochemistry data a slightly, but statistically significantly increased uric acid and total bilirubin level in males (p<0.05 - p<0.01), and slightly increased chloride level in females (p<0.05) were yet to be noted.

Subsequently, no further plasma discoloration was observed, whereas urinalysis indicated a deep-yellow urine discoloration in all males and in two out of five females.

ORGAN WEIGHTS AND ORGAN WEIGHT RATIO

There were no treatment-related changes in organ weights and ratios.

MACROSCOPIC AND MICROSCOPIC FINDINGS

At the end of the dosage period, minor morphological alterations indicative of a slight irritant effect of the test article, were present in the gastric mucosa of rats of group 4 and to a lesser degree in group 3. These alterations took the form of minimal to slight degrees of apoptotic bodies in parietal cells accompanied by vacuolation of limiting ridge epithelium and inflammatory cell infiltration of the glandular submucosa. These findings had largely reversed following the recovery period.

DISCUSSION

The oral administration of SCARLET RN 1165 to Wistar rats at doses of 50, 200 and 1000 mg/kg/day, for 28 days, resulted in no effects on mortality, food consumption, body weight, ophthalmoscopic examinations and organ weights.

Treatment with test article , a pigment, resulted in red feces in males at 200 mg/kg. Red feces and red urine was observed in all animals at 1000 mg/kg.

Test article-related changes of clinical biochemistry and urinalysis comprised the following parameters: slightly decreased glucose level in females of group 4 (p<0.01), marginally increased creatinine level in males of group 4 (p<0.05), slightly increased uric acid level in both sexes of group 4 (p<0.01), slightly to markedly increased total bilirubin level in both sexes of groups 3 and 4 (p<0.05 - p<0.01), slightly increased triglyceride level in both sexes of group 4 (p<0.01), slightly decreased calcium level (p<0.01) and marginally increased chloride level (p<0.05) in females of group 4.

Plasma discoloration and urine discoloration was observed in all animals of groups 3 and 4. Urine discoloration was also present in some animals of group 2 (50 mg/kg)

Recovery period - all of the findings recorded during the treatment period were found to be reversible and generally similar to those of the controls. However, for clinical biochemistry data a slightly, but statistically significantly increased uric acid and total bilirubin level in males (p<0.05 - p<0.01), and slightly increased chloride level in females (p<0.05) were yet to be noted.

Subsequently, no further plasma discoloration was observed, whereas urinalysis indicated a deep-yellow urine discoloration in all males and in two out of five females.

Reddish discoloration was noted mainly in the digestive tract and kidneys from primarily rats of group 4 and 3 at the end of the main study period. This was due to the test article, a dyestuff and was not accompanied by microscopic alterations.

Morphologic alterations following the administration of the test article were present in the stomach.

These took the form of minor degrees of apoptotic bodies in parietal cells accompanied by also minor degrees of vacuolation of limiting ridge epithelium and inflammatory cell infiltration of the glandular submucosa. These findings had largely reversed following the recovery period. Both sexes of group 4 (1000 mg/kg) were clearly affected, group 3 (200 mg/kg) to a lesser degree. These findings may be regarded as indicators of a slight irritant effect of the test article on the gastric mucosa.

CONCLUSION

Oral administration of SCARLET RN 1165 to Wistar rats at doses of 0, 50, 200 and 1000 mg/kg/day, performed according to OECD 407/method EC B7, for 28 days resulted in the following:

Based on the results of this study, 50 mg/kg SCARLET RN 1165 was established as the no-observed-adverse-effect level (NOAEL).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
50 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Klimish code 1 (OECD 407 & GLP)
System:
gastrointestinal tract
Organ:
other: gastric mucosa

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification