Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
other: Crl : CD (SD)
Sex:
male/female

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Test duration: 28 days
Frequency of treatment:
Once daily for consecutive 28 days period.
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Male: 5 animals at 0 mg/kg bw/day
Male: 5 animals at 100 mg/kg bw/day
Male: 5 animals at 300 mg/kg bw/day
Male: 5 animals at 1000 mg/kg bw/day
Female: 5 animals at 0 mg/kg bw/day
Female: 5 animals at 100 mg/kg bw/day
Female: 5 animals at 300 mg/kg bw/day
Female: 5 animals at 1000 mg/kg bw/day
Control animals:
yes

Examinations

Observations and examinations performed and frequency:
Mortality/moribundity: Twice daily at least at six hours apart during study period.
Cage side clinical observation: Once daily during study period.
Detailed clinical observation: on randomization day and once weekly during study period.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
no mortality occurred and no test article-related clinical signs were identified.
Mortality:
no mortality observed
Description (incidence):
No mortality occurred and no test article-related clinical signs were identified through out the study period. Marked swelling, erythema on right hind paw and lameness was observed in one male animal (ID: 1110411016) at 1000 mg/kg/day and were considered as accidental injury by its cage and was consistent with microscopic findings. Hair loss was observed in two females at 100 mg/kg/day and wound on right forepaw was observed in one female at 1000 mg/kg/day. These above two clinical signs were considered from over self-grooming of individual housing animals.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No treatment related body weight changes were observed.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not specified
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No definitely test article-related changes in serum biochemistry parameters were noted. Statistically decreased alkaline phosphatase was observed in males at 100 mg/kg/day and higher, and was still within the historical data range. The physiological functions of animals were normal. Higher aspartate aminotransferase and potassium were observed in females at 300 and 1000 mg/kg/day, respectively. The findings had no dose-dependence and also still within the historical data range. These above changes considered as incidental findings and were not adverse effects.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
The test artical-related changes were the orange-colored urine, positive response in bilirubin and glucose observed in male and female rats at 1000 mg/kg/day. These findings may relate to renal function change but there were no other correlated findings in serum biochemistry parameters such as blood urea nitrogen or creatinine. The findings in urine were not considered as adverse effect.
In urine sediment examination, occasional RBCs were noted in some animals. These findings without dose relationship were considered as animal individual difference.
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Abnormal contents were noted in one or more of the intestinal segments (colon, cecum or ileum) in male and female rats at 1000 mg/kg/day and considered related to the administration of the test article. It was, however, attributed to the residual unabsorbed test article within the intestinal tract instead of a genuine treatment effect. This finding was not traceable microscopiacally for correlation. A male at 1000 mg/kg/day had dark red discoloration in the dorsal aspect of the right foot. This correlated with hemorrhage/edema with hyperplasia of the epidermis and serocellular crust microscopically and was compatible with injury caused by mishandling (handling accident).

None of the microscopic changes observed were considered treatment-related. A slight increase in the incidence of minimal mononuclear cell infiltration in the liver was noted in the male and female rats at 1000 mg/kg/day. However, this was a common background finding in laboratory rats and the biological/toxicological significance of this was questionable due to its low severity. Pyogranulomatous pneumonia was noted in a single male at 1000 mg/kg/day and was attributed to aspiration of the test article.
Histopathological findings: neoplastic:
no effects observed

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
histopathology: non-neoplastic

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1.  Mortality and Clinical Signs
Items   Dose level   (mg/kg/day)   Total incidence (n/n)
Male                   
Mortality 0a 0/5
100 0/5
300 0/5
1000 0/5
Clinical signs
Swelling on hind paw, 0a 0/5
Lameness, 100 0/5
Erythema on hind paw 300 0/5
1000 1/5
Female         
Mortality 0a 0/5
100 0/5
300 0/5
1000 0/5
Clinical signs
Hair loss 0a 0/5
100 2/5
300 0/5
1000 0/5
Wound 0a 0/5
100 0/5
300 0/5
1000 1/5
         
a: WFI
n/n: Total number of abnormal or dead animals observed/Total number of animals examined

Applicant's summary and conclusion

Conclusions:
No adverse effects were observed in clinical observation, body weight, food consumption, clinical pathology, macro- and microscopic examinations up to the dose of 1000 mg/kg/day. The NOAEL of Everzol Orange ED-G Crude is determined at dose level of 1000 mg/kg/day under the condition of this study.
Executive summary:

No mortalities were observed in male and female rats dosed with Everzol Orange ED-G at 100, 300 or 1000 mg/kg/day. There were no test article treatment-related changes in clinical observation, body weight, food consumption, hematology, coagulation and serum biochemistry. The test article-related changes were limited to urinalysis which included the orange-colored urine, positive responses in bilirubin and glucose observed in male and female rats at 1000 mg/kg/day. However, there was no other correlated finding in serum biochemistry. The physiological function of animals was normal. These findings in urine were not considered as adverse effects.

At the end of study, there were no treatment-related changes in organ weight or microscopic changes were noted. Grossly, abnormal contents were noted in one or more intestinal segments (colon, cecum or ileum) of male and female rats at 1000 mg/kg/day. However, this was attributed to the residual unabsorbed test article within the intestinal tract instead of a genuine treatment effect. The physiological function of animals was still normal. The gross finding of animals was not considered as adverse effects.

In conclusion, no adverse effects were observed in clinical observation, body weight, food consumption, clinical pathology, macro- and microscopic examinations up to the dose of 1000 mg/kg/day. The NOAEL of Everzol Orange ED-G Crude is determined at dose level of 1000 mg/kg/day under the condition of this study.