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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
no data
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: well-conducted study but reported in Japanese language
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1975

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
A combination of rules from OECD Guidelines 409 and 452 were followed (by anticipation as thoses guidelines did not exist when the study was conducted).
d-limonene was administered by gavage to dogs for 6 months. This duration was chosen as usual when developing new medicinal products (d-limonene was explored in this study as a possible gallstone solubiliser).
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
no data

Test animals

Species:
dog
Strain:
other: Japanese beagle
Sex:
male/female

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
no data
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
6 months
Frequency of treatment:
once a day
Doses / concentrations
Remarks:
Doses / Concentrations:
0.4, 1.2 and 3.6 mL/kg bw/d equivalent to 340, 1000 and 3000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
3/sex/dose
Control animals:
yes

Results and discussion

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Decreased body weight and protein casts observed in the renal tubule at 3000 mg/lg bw/d.
Dose descriptor:
LOAEL
Effect level:
3 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Dose descriptor:
NOAEL
Effect level:
340
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Decreased body weight and protein casts observed in the renal tubule at 1000 mg/lg bw/d.
Dose descriptor:
LOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

- Clinical signs: frequent vomiting and nausea were caused, which appeared to depend on the dose used.

- Body weight: all males in the high dose group, all females in the intermediate group and 2/3 females in the high dose group lost weight over the 6 month-study. Not dose-related in females.

- Food consumption: not affected by treatment except in the intermediate dose group females. Not dose-related.

- Urinalysis: no treatment-related effects.

- Hematology: no treatment-related findings.

- Biochemistry: total cholesterol and glucose decrease in blood in the high dose group. The initial cholesterol level was recovered at the end of the 6-month treatment period in both sexes (when the level had increased in other control and treated groups).

- Organ weights: the relative to body weight kidney and liver weights were slightly higher in the high dose group males than in other groups.

- Histopathology: at 1.2 and 3.6 mL/kg bw/day, protein casts were observed in the renal tubule of most animals. No remarkable treatment-related change was observed in other organs.

Table 1: Protein casts observed in the renal tubule

  Male Female
  Control 0.4 1.2 3.6 Control 0.4 1.2 3.6
No. animals examined 3 3 3 3 3 2 3 3
Protein casts 1 1 2 3 1 2 3 3

Applicant's summary and conclusion

Conclusions:
The NOAEL in this study is 1.2 mL/kg bw/day (equivalent to 1000 mg/kg bw/day) in males and 0.4 mL/kg bw/day (equivalent to 340 mg/kg bw/day) in females on the basis of decreased body weight and protein casts observed in the renal tubule at the next dose level. Food consumption was also decreased in the intermediate dose females.
Executive summary:

Three dogs per sex and per dose group were administered d-limonene by gavage once per day for 6 months at the dose level of 0, 0.4, 1.2 or 3.6 mL/kg bw/day.

All 6 animals (males and females) from the high dose group except one female and all females in the intermediate dose group lost weight between the first and the last day of study. Food consumption only decreased in the intermediate dose group females. Urinalysis and hematology were not affected by treatment. The glucose and total cholesterol levels in blood decreased in the high dose group males and females when compared to the pre-treatment levels; the total cholesterol level recovered the pre-test level by the end of the 6-month treatment period. The relative kidney and liver weights were slightly higher in the high dose group males than in other groups. A dose-related increased incidence of protein casts were observed in the renal tubule: all males in the high dose group and all females in the intermediate and high dose groups showed this effect.