Morpholinium, 4-[2-[2-[(2-hydroxyphenyl)methylene]hydrazinyl]-2-oxoethyl]-4-methyl-, chloride (1:1)

Endpoint summary

Description of key information

Based on the results of a combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test of TINOCAT ES 96000 in rats by oral gavage, the changes noted for females at the highest dose level were considered to be treatment related but not adverse. Therefore, a NOAEL of 800 mg/kg bw/day was derived.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

A GLP compliant Combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test was performed according to guideline OECD 422 (WIL Research Europe B.V. 2013). After acclimatization, four groups of ten male and ten female Wistar Han rats were exposed daily by oral gavage to the test substance,Tinocat ES 96000, at 0, 80, 250 and 800 mg/kg bw/day (dose level selection based on the results of a 14-day dose range finding study). Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 43- 54 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation. Formulation analysis showed that the formulations were prepared accurately, were homogenous, and were stable for at least 5 hours at room temperature. Females at 800 mg/kg bw/day had higher absolute and relative spleen weights. Microscopic alterations associated with treatment were present in the spleen and sternal bone marrow of females. These were characterized by an increased severity of primarily erythroid hemopoeitic foci in the spleen of treated animals and a parallel increase in the incidence of minimal erythroid hyperplasia in the bone marrow. However, this alteration may have reflected a slightly increased erythrocyte turnover in treated animals, which remained within physiological limits. Taken together, the changes noted for females at 800 mg/kg bw/day were considered to be treatment related but not adverse.In conclusion, treatment with Tinocat ES 96000 by oral gavage in male and female Wistar Han rats at dose levels of 80, 250 and 800 mg/kg bw/day revealed no toxicity up to 800 mg/kg bw/day. Based on these results, a No Observed Adverse Effect Level (NOAEL) of 800 mg/kg bw/day was derived.

Range-finder

A 14-Day dose range finding study for a combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test of Tinocat ES 96000 was conducted in rats by oral gavage.

Guidelines

No guidelines are applicable as this pilot study was used to select dose levels for the Combined 28-day Repeated Dose Toxicity Study with the Developmental/Repro Screening Test of Tinocat ES 96000 (Project 501296; BASF Project 85R0013/11X545).

Rationale for dose levels

After single oral administration of Tinocat ES 96000 three out of 6 rats died at 2000 mg/kg bw/day. No animals died at 300 mg/kg bw/day. The dose levels for this 14-day toxicity study were selected to be 0, 300 and 800 mg/kg bw/day.

Study outline

After acclimatization, three groups of four male and four female Wistar Han rats were exposed by oral gavage to the test substance at 300 (Group 2) or 800 mg/kg bw/day (Group 3) for 14 days. Rats of the control group (Group 1) received the vehicle, 1% Aqueous carboxymethyl cellulose, alone.

Evaluated parameters

The following parameters were evaluated: mortality / viability, clinical signs, body weights, food consumption, clinical laboratory investigations (haematology, clinical biochemistry), macroscopy and organ weights. Chemical analyses of formulations were conducted once during the study to assess accuracy, homogeneity and stability.

Results/discussion

Formulation analysis showed that the formulations were prepared accurately and homogenously, and were stable when stored at room temperature under normal laboratory light conditions for at least 5 hours.

Results:

Effects related to treatment with Tinocat ES 96000 included lower body weight gain with decreased food intake at 800 mg/kg bw/day, and to a lesser extent also at 300 mg/kg bw/day. A slight increase in aspartate aminotransferase was seen in males at 800 mg/kg bw/day, as well as slightly lower liver weights. No changes were noted in clinical appearance, haematology investigations and macroscopic examination up to a dose of 800 mg/kg bw/day. Based on the results of this dose range finding study, dose levels of 0, 80, 250 and 800 mg/kg bw/day were selected for the main study (Project 501296; BASF Project 85R0013/11X545), taking into account the longer administration period and inclusion of pregnant animals. Due to the absence of clinical signs after dosing in the present study, clinical observations and functional observational tests were conducted directly after dosing in the main study.

Justification for classification or non-classification

Based on the available data, the test substance is not classified with regard to repeated dose toxicity according to Directive 67/548/EEC (DSD) and Regulation (EC) No 1272/2008 (CLP), respectively.