A mixture of: 4-(2,2,3-trimethylcyclopent-3-en-1-yl)-1-methyl-2-oxabicyclo[2.2.2]octane; 1-(2,2,3-trimethylcyclopent-3-en-1-yl)-5-methyl-6-oxabicyclo[3.2.1]octane; spiro[cyclohex-3-en-1-yl-[(4,5,6,6a-tetrahydro-3,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]furan]; spiro[cyclohex-3-en-1-yl-[4,5,6,6a-tetrahydro-4,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]]furan]

Endpoint summary

Description of key information

28 -day repeated dose (oral) (OECD TG 407): The NOAEL was determined to be ≥1000 mg/kg bw/day for males and females, as no adverse effects were observed at a dose of 1000 mg/kg/day. This value will be used for the risk assessment.

Reproscreening test (OECD TG 421): Treatment with the substance by dietary administration in male and female Wistar Han rats at dose levels of 1.000, 2.500 and 8.000 ppm (circa 70, 170 and 550 mg/kg bw/day) revealed slight non-adverse female toxicity at 550 mg/kg bw/day. The NOAEL was determined to be >550 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The database consists of 2 studies which are carried out according to current guidelines which confirm the results found and therefore the quality of the database is high.

Additional information

28 -day repeated dose (oral) (OECD TG 407)

In a 28 -days repeated dose toxicity study (OECD TG 407, GLP) with a 2 -week recovery period, the substance was administered via oral gavage to Sprague Dawley rats (five males and five females) at dosages of 15, 150 and 1000 mg/kg/day. The substance was prepared as a suspension in corn oil at concentrations of 0.3, 3.0 or 20% w/v. Control animals (five males and five females) received the vehicle (corn oil) only, at the same dose volume (5mL/kg/day). All parameters from OECD TG 407 were recorded. Blood samples for clinical investigations were taken on day 27 and all animals were killed and examined macroscopically on day 29. Histopathological examination of the tissues was then initiated. The following results were found: Clinical signs: no adverse effects considered to be associated with treatment were observed for mortality, clinical signs, bodyweight, food consumption or efficiency of food utilisation. Haematology: no effects observed. Biochemical parameters: lower than control glucose levels were recorded specifically for male and to a lesser extent for female rats receiving 1000 mg/kg/day. Higher cholesterol levels for male rats receiving 1000 mg/kg/day were seen. Organ weights: higher than control liver weights (bodyweight adjusted) were recorded for male and female rats treated at 1000 mg/kg/day (36 and 47%, respectively). Kidney weights were higher than control for male rats treated at 1000 mg/kg/day (ca 10%). Macroscopically no effects were seen. Microscopic pathology: Minimal centrilobular hepatocyte enlargement (males) and minimal generalised hepatocyte enlargement (females) were seen in the liver of rats receiving 1000 mg/kg/day. A marginally increased incidence and degree of eosinophilic inclusions were seen in the proximal tubular epithelium of the kidneys of male rats receiving 1000 mg/kg/day. The changes in the liver are considered to be of an adaptive nature due to administration of the test substance which is readily absorbed and are therefore not considered for the derivation of the NOAEL. The minor kidney effects are related to alpha-hydrocarbon nephropathy and are not taken into account for deriving the NOAEL for man because this is a male rat specific phenomenon. Based on the findings, the NOAEL was determined to be ≥1000 mg/kg bw/day for males and females.

Reproscreening study (OECD TG 421)

In a dietary reproscreen study according to OECD TG 421 some repeated dose toxicity was also considered. the doses were 8000, 2500 and 1000 ppm, equivalent to 550, 175 and 70 mg/kg bw. The doses were based on the OECD TG 407 (1000 mg/kg bw as the highest dose) and the 14-day DRF 10000, 3000 and 1000 ppm (eq to ca 1000, 300 and 100 mg/kg bw. Based on clinical signs seen in the OECD TG 407 and increased liver weights >> 20% both in the OECD TG 407 (accompanied with liver hypertrophy) and this DRF the doses were slightly lowered in the main OECD TG 421 test. The fertility and developmental toxicity parameters are presented in the subsequent sections. Body weight (gain), clinical signs, food consumption and some organs were evaluated. No effects were seen on body weight (gain) and food consumption. Some piloerection was seen in the females. Relative liver weights were increased in males and females in the high dose (+20 and +30%, respectively) and minimal liver hepatocellular hypertrophy was noted at minimal degree and was considered to be an adaptive non adverse finding. Adrenal weight were decreased but no macroscopic or microscopic findings related to treatment were noted. The NOAEL for males and females for repeated dose effect is considered to be 550 mg/kg bw/day.

Justification for classification or non-classification

Based on the results of the 28 -day repeated oral gavage toxicity study (OECD TG 407) and diet reproscreening study (OECD TG 421), the substance does not have to be classified for repeated dose toxicity according to EU CLP (EC 1272/2008 and its amendments).