Registration Dossier

Administrative data

Description of key information

Oral:
- NOAEL (local iritation effects) = 50 mg/kg bw/day (Wistar rat, OECD TG 422, 30 (males) - 49 (females) days, gavage)
- NOAEL (general, systemic toxicity) = 150 mg/kg bw/day (Wistar rat, OECD TG 422, 30 (males) - 49 (females) days, gavage)
Dermal:
- no data
Inhalation:
- no data

Key value for chemical safety assessment

Additional information

Oral:

Glycerin 3 EOTA was tested in a Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test according to OECD TG 422 and in compliance with GLP regulations by BASF SE (2010). The substance was administered orally via gavage to groups of 10 male and 10 female Wistar rats (F0 animals) at doses of 50, 150, and 500 mg/kg body weight/day. Control animals were dosed daily with the vehicle (1% Carboxymethylcellulose suspension in drinking water). The duration of treatment covered a 2-week pre-mating and mating period in both sexes, approximately 1 week post-mating in males, and the entire gestation period as well as 4 days of lactation in females.

 

After 2 weeks of premating treatment F0 animals were mated to produce F1 generation pups. Mating pairs were from the same test group. Mating was discontinued as soon as sperm was detected in the vaginal smear. F0 animals were examined for their reproductive performance including determination of the number of implantation sites and the calculation of postimplantation loss for all F0 females. A detailed clinical observation (DCO) was performed in all animals before test substance administration and thereafter at weekly intervals. Food consumption of the F0 parents was determined once weekly during premating. In dams food consumption was determined for gestation days 0-7, 7-14, 14-20 and lactation days 1-4. Body weights of F0 parents were determined once a week, in males throughout the study and in females during premating. During gestation and lactation period, F0 females were weighed on gestation days (GD) 0, 7, 14 and 20, on the day of parturition (postnatal day [PND] 0) and on PND 4. Pups were sexed on PND 0 and weighed one day after birth and on PND 4. Their viability was recorded twice daily on each workday or only in the morning on Saturday and Sunday. On PND 4 all pups were sacrificed under Isoflurane anesthesia with CO2and were examined macroscopically. Clinicochemical and hematological examinations as well as urinalyses were performed in 5 parental males and females per group towards the end of the administration period. At the end of the administration period a functional observational battery was performed and motor activity was measured in 5 parental males and females per group. All surviving F0 parental animals were sacrificed by decapitation, under Isoflurane anesthesia, and were assessed by gross pathology. Organ weights were recorded and a histopathological examination was performed.

 

Clinically, distinct toxicity was noted in the F0 males and F0 females at 500 mg/kg bw/d. Mortality in 4 high-dose males and 3 high-dose females as well as clinical observations such as respiratory sounds, labored respiration and piloerection were observed during major parts of treatment. These findings were also observed in individual males at 150 mg/kg bw/d, however, they were less severe and no mortality was observed there. Detailed clinical examinations in an open field and in a functional observational battery (FOB) confirmed the findings made during cage-side observations, but measurements of motor activity did not reveal any indications of test substance-induced effects in low-, mid- and high-dose rats (50, 150 and 500 mg/kg bw/d).

 

Pathology (see below) confirmed that mortality and adverse clinical observations were subsequent to local irritant effects of the test substance in the fore- and glandular stomach. Also, reduced food consumption in males and females during premating and reduced body weights/body weight gain in males are considered to be secondary to these local effects. Thus, these distinct adverse effects, in particular mortality, were not regarded to be due to systemic toxicity of Glycerin 3 EOTA.

 

All high-dose as well as most mid-dose animals of both sexes showed transient salivation for a few minutes immediately after each treatment. This was likely to be induced by the unpleasant taste of the test substance and/or by local irritation of the upper digestive tract. It was neither considered to be a sign of systemic toxicity nor as adverse.

 

Clinical chemistry revealed a slight anemia in mid- and high-dose females (150 and 500 mg/kg bw/d) indicated by reduced RBC counts, haemoglobin and hematocrit values. Maybe, anemia was due to erosions and ulcers in the forestomach and glandular stomach of rats of both sexes in the mentioned dose groups. Females seemed to be more prone to anemia due to gestation and lactation. The increased synthesis of coagulation factors (i.e., reduced prothrombin time) in males and females of test group 3 (500 mg/kg bw/d) as well as the increased platelet counts in females of test groups 2 and 3 (150 and 500 mg/kg bw/d) happened consequently to the erosions in the stomach. Most probably, higher cholesterol levels in rats of both sexes of test group 3 (500 mg/kg bw/d) as well as higher triglyceride levels in females of the test groups 2 and 3 (150 and 500 mg/kg bw/d) were due to an altered liver cell metabolism induced by the substance administration. The lower alkaline phosphatase activity in males of test group 3 (500 mg/kg bw/d) was most probably combined to the reduced body weights.

 

Pathology and histopathology revealed fore- and glandular stomach, liver lymph node, duodenum, and liver as target organs in this study.

 

In the forestomach, erosions or ulcers were noted in 7 males and one female of test group 2 (150 mg/kg bw/day) as well as in all males and females of test group 3 (500 mg/kg bw/day). The erosions or ulcers were associated with inflammation. Their occurrence is regarded to be the primary response to an irritant effect of the test substance. As proliferative response secondary to ulceration, squamous cell hyperplasia was observed in 8 males and one female of test groups 2 (150 mg/kg bw/day) as well as in all males and 9 females of test group 3 (500 mg/kg bw/day). The severity of the squamous cell hyperplasia was dose-related increased. In test group 3 (500 mg/kg bw/day), all decedents showed erosions or ulcers and squamous cell hyperplasia in the forestomach. The occurrence of erosions or ulcers and squamous cell hyperplasia was considered to be treatment-related and adverse.

 

In the glandular stomach, the test substance led to erosions or ulcers that were observed in 5 males and 2 females of test group 3 (500 mg/kg bw/day). In addition, focal hyperemia was noted in 4 males and 2 females of test group 3 (500 mg/kg bw/day). Erosions or ulcers and the focal hyperemia were considered as consequence of irritant effects of the test substance and were regarded to be adverse.

 

In the liver lymph node, a lympho-reticulocellular hyperplasia was observed in 3 males of test group 2 (150 mg/kg bw/day) as well as in 7 males and 7 females of test group 3 (500 mg/kg bw/day). Furthermore, lymphoid cysts occurred in 5 males and 5 females of test group 3 (500 mg/kg bw/day). These findings were considered to be secondary effects as consequence of the inflammatory processes in the forestomach and regarded to be adaptive and non-adverse.

 

In the duodenum, a diffuse thickening of duodenal mucosa was noted in 9 males and 7 females of test group 3 (500 mg/kg bw/day). Because the histological structures of the duodenum in the affected animals were comparable to controls and because the severity was only minimal or slight, this finding was considered to be adaptive and non-adverse.

 

The increased liver weights in females of test group 2 (150 mg/kg bw/day) as well as in males and females of test group 3 (500 mg/kg bw/day) are related to treatment. They are regarded to be adaptive and non-adverse. In males, the increased liver weights correlated with a minimal central hepatocellular hypertrophy that occurred in the liver of 5 animals in test group 3 (500 mg/kg bw/day). The hepatocellular hypertrophy was assessed to be treatment-related and adaptive.

 

In conclusion, under the conditions of the present reproduction/developmental toxicity screening test the NOAEL (no observed adverse effect level) for general, systemic toxicity of the test substance was 150 mg/kg bw/d for males and females based on clinical chemistry changes at next higher dose level. Local irritation in fore- and glandular stomach led to mortality at 500 mg/kg bw/d and triggered adverse clinical observations as well as hematological changes at 150 and 500 mg/kg bw/d. Thus, the NOAEL for local irritation effects was equivalent 50 mg/kg bw/d.

 

Dermal:

No data available.

 

Inhalation:

No data available.

Justification for classification or non-classification

Under the conditions of the present reproduction/developmental toxicity screening test the NOAEL (no observed adverse effect level) for general, systemic toxicity of the test substance was 150 mg/kg bw/d for male and female rats based on clinical chemistry changes at the next higher dose level (500 mg/kg bw/d). Adverse clinical observations as well as hematological changes seen at 150 mg/kg bw/d were assessed to be secondary effects to the local irritation observed in the fore- and glandular stomach. Local irritation effects towards mucous membranes are already covered by the substance's classification for eye and respiratory tract irritation after single exposure (R41 / Eye irritation category 1 and R37 / STOT SE category 3). Accordingly, local effects are not taken into consideration for classification concerning specific target organ toxicity after repeated exposure. Based on the LOAEL for systemic toxicity of 500 mg/kg bw/day, the substance does not have to be classified for repeated dose toxicity.

Thus, based on the available data, no classification of the substance concerning repeated dose toxicity is warranted.

  • EU classification according to Annex VI of Directive 67/548/EEC: no classification required
  • GHS classification (REGULATION (EC) No 1272/2008 (CLP)): no classification required