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Description of key information

Macrolex Rot EG was administered by gavage in doses of 0, 100, 300, 1000 mg/kg bw/day suspended in polyethylene glycol 400 to male and female Wistar rats for a period for 4 weeks according to OECD TG 407 and GLP. Under the conditions described the NOAEL for oral administration of Macrolex Rot EG to male and female rats is 1000 mg/kg bw/day 

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:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: guideline study and GLP
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMAL
- Age at study initiation: 5 weeks
- Weight at study initiation: male 173.6 g; female: 145.3 g
- Housing: in groups of 2 or 3
- Diet ad libitum
- Water ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 55
- Air changes (per hr): >10 per hour
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
The test item was administered by gavage to male and female Wistar rats in daily doses of 100, 300 and 1000 mg/kg bw/day suspended in polyethylene glycol 400. The administration volume was 5ml/kg bw.
Details on analytical verification of doses or concentrations:
The formulation was prepared as needed taking into account the analytically determined stability of 8 days
Duration of treatment / exposure:
28 day
Frequency of treatment:
once daily
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw/day
Basis:

No. of animals per sex per dose:
5 per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
The test item was administered once daily by gavage to male and female Wistar rats in daily doses of 100, 300 and 1000 mg/kg bw/day suspended in polyethylene glycol 400. The administration volume was 5 ml/kg bw. the animals were observed for mortality and clinical signs, determination of hematology and clinical chemistry data. After termination of treatment animals were subjected to gross and histopathological examination
Positive control:
no
Observations and examinations performed and frequency:
Inspection of Animals for Morbidity and Mortality twice daily
General Clinical Observations (in cage) daily
Detailed Clinical Observations including
Open Field Observation (OFO) weekly
Determination of:
Body Weight(s) daily
Food Consumption weekly
Water Consumption weekly
Clinical Pathology:
---Hematology; day 29, 30
Differential blood count, erythrocyte morphology, erythrocyte count, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean corpuscular volume, hemoglobin concentration, hematocrit, leukocyte count, reticulocyte count, thrombocyte count, thromboplastin time (Hepato-Quick).
----Clinical Chemistry; day 29, 30
Alanine aminotransferase, alkaline phosphatase, albumin, total bilirubin, cholesterol, creatinine, total protein, urea, glucose, potassium, sodium, gall acids.
Functional Observation Battery: day 23, 24
Motor/Locomotor Activity: day 23, 24
Sacrifice and pathology:
Necropsy: day 29, 30
determination of organ weight and calculation of relative organ weight:
Brain, heart, liver, spleen, kidneys (both), thymus, adrenal glands (both), epididymides (both), testes (both), prostate, seminal vesicles (with coagulation glands), ovaries/oviducts (both) and uterus/cervix.
gross and histopathological examination
all weighed organs and additionally
caecum, colon, duodenumeyes, femur with bone marrow, joint, ileum, jejunum, larynx, lungs, lymph nodes mandibular mesemteric iliacale, optic nerves, pancreas, Peyer's patches, rectum, siatic nerve
Statistics:
Dunnett, U-test, Het-Dunn ( Dunnett exact test heterogeneous test)
Details on results:
---Mortality
Survival was not affected in all dose groups
---Clinical findings
No findings in detailed clinical observations were observed
In-cage observations revealed changes in feces color (reddish particles) in males and females of the 300 mg/kg bw. dose group starting on day 3 and in the 1000 mg/kg bw dose group starting on day 2 up to termination.
---Body weight development
Body weights were not affected by the treatment with Macrolex Rot EG at doses up to 1000 mg/kg bw
---Hematology
Red and white blood parameters including blood coagulation were not toxicologically relevantly changed up to 1000 mg/kg bw..
---Clinical Chemistry
Clinical chemistry revealed a decrease of alkaline phosphatase (APh) in dosed females starting at 100 mg/kg bw. and being statistically significant at 300 and 1000 mg/kg bw. A decrease in APh is not considered as toxicologically relevant. All other parameters including electrolytes were in the normal range
---Organ weights
No toxicological relevant changes in absolute and relative organ weights could be observed.
---Gross and histopathological evaluation
At necropsy, there was no evidence of any gross finding that had to be attributed to dosing with the test compound.
Histopathology revealed minimally or slightly reduced secretion in the seminal vesicles in 3 out of 5 males at 1000 mg/kg bw. which was not associated with degeneration. Changes in other male genital organs (testes, epididymides, prostate or coagulation glands) could not be found up to and including 1000 mg/kg bw. Thus, reduced secretion is regarded as unspecific and possibly stress-related. An adverse effect is not assumed.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: No relevant adverse effects could be observed
Critical effects observed:
not specified
Executive summary:

Macrolex Rot EG was administered by gavage to 5 male and 5 female Wistar rats per dose group indayily doses of 0, 100, 300, 1000 mg/kg bw/day suspended in polyethylene glycole 400 (vehicle) for a period of 28 days. The study was concucted in compliance with OECD TG 407 under GLP conditions.

Survival was not affected by treatment with Macrolex Rot EG and no clinical signs were observed. The observed decreased alkaline phosphatase in females was not considered relevant because neither all other clinical chemistry data nor hematology gave evidence for treatment related effects up to 1000 mg/kg bw. Starting at the mid dose of 300 mg/kg bw,. all animals of both sexes showed changes in feces color (reddish particles), which is a consequence of the strong red color of the test item and thus is not considered as adverse effect. Histopathology revealed minimally to slightly reduced secretion in the seminal vesicles in 3 out of 5 males at 1000 mg/kg bw which was not associated with degeneration of seminal vesicles. Changes in other male genital organs (testes, epididymides, prostate or coagulation glands) could not be found up to and including 1000 mg/kg bw.. Therefore, reduced secretion is regarded as unspecific and possibly stress-related. An adverse effect is not assumed. Female reproductive organs were not affected up to 1000 mg/kg bw/day.

In conclusion, under the conditions described above, the no-adverse-effect-level (NOAEL) for male and female rats after 4 -week daily oral treatment by gavage with Macrolex Rot EG is 1000 mg/kg bw/day.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
A reliable study which was performed according to OECD TG 407 and GLP with Klimisch score 1

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

Additional information

ORAL APPLICATION

Macrolex Rot EG was administered by gavage to 5 male and 5 female Wistar rats per dose group in daily doses of 0, 100, 300, 1000 mg/kg bw/day suspemnded in polyethylene glycol 400 (vehicle) for a period of 28 days. The study was conducted in compliance with OECD TG 407 under GLP conditions.

Survival was not affected by treatment with Macrolex Rot EG and no clinical signs were observed. The observed decreased alkaline phosphatase in females was not considered relevant because neither all other clinical chemistry data nor hematology gave evidence for treatment related effects up to 1000 mg/kg bw. Starting at the mid dose of 300 mg/kg bw,. all animals of both sexes showed changes in feces color (reddish particles), which is a consequence of the strong red color of the test item and thus is not considered as adverse effect. Histopathology revealed minimally to slightly reduced secretion in the seminal vesicles in 3 out of 5 males at 1000 mg/kg bw which was not associated with degeneration of seminal vesicles. Changes in other male genital organs (testes, epididymides, prostate or coagulation glands) could not be found up to and including 1000 mg/kg bw.. Therefore, reduced secretion is regarded as unspecific and possibly stress-related. An adverse effect is not assumed. Female reproductive organs were not affected up to 1000 mg/kg bw/day.

In conclusion, under the conditions described above, the no-adverse-effect-level (NOAEL) for male and female rats after 4 -week daily oral treatment by gavage with Macrolex Rot EG is 1000 mg/kg bw/day.

There is no sub-chronic study available as required by Regulation (EC) No.1907/2006 (REACH) ANNEX IX, section 8.6. According to ANNEX IX, Section 8.6 Column 2 of Regulation EC (REACH) No 1907 (2006) this sub-chronic study needs not to be conducted because the substance is unreactive, insoluble and not inhalable and there is no evidence of absorption and no evidence of toxicity in a 28 -day "limit test". In the available 28-day study (OECD TG 407 and GLP) male and female rats were given daily doses up to and including 1000 mg/kg bw/day (limit dose). Based on the data generated from this study no relevant adverse effects of the substance were found up to and including 1000 mg/kg bw/day. Therefore the NOAEL (No Observed Adverse Effect Level) of Macrolex Rot EG is considered to be 1000 mg/kg bw/day. The NOAEL -28days allows the extrapolation towards a NOAEL-90 days for the same route of exposure by application of an appropriate uncertainty factor. Furthermore, as no evidence of relevant adverse effects up to the limit dose of 1000 mg/kg bw/day are reported, and it can be assumed that prolongation of treatment time does not provide additional relevant information on toxic effects.

Overall, there is a fully valid study available in which rats tolerate doses up to the limit dose of 1000 mg/kg bw over a period of four weeks without mortality and without relevant adverse effects, thus providing evidence that prolongation of treatment period do not lead to additional relevant adverse effects in rats. Consequently, the NOAEL of 1000 mg/kg/day is taken as a starting point for DNEL calculation and a conservative assessment factor for time extrapolation from sub-acute to chronic exposure (AF 6) is applied.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
A reliable study which was performed according to OECD TG 407 and GLP with Klimisch score 1

Justification for classification or non-classification

Based on the available data no classification or labelling is required.