Reaction mass of 1,3-dioxan-5-ol and 1,3-dioxolan-4-ylmethanol

Repeated dose toxicity: other routes

Administrative data

Endpoint:

short-term repeated dose toxicity: other route

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The route of administation is of low relevancy for a REACH dossier. Raw data of body weight changes are not available. Statistical study was performed but the method is not presented. 28-day repeated toxicity tests (like OECD TG 407) uses 3 dose levels (and control) and at least 5 males and 5 females per group. In this test, 2 dose levels (and control) and 5 animals per group were used.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Toxicity tests after intravenous (i.v.) application of glycerol formal was performed with rabbits for a period of 4 weeks.
The test item was applied daily as 30% dilution with 0.9% NaCl-solution at dose of 0.024 ml/kg (dose I) and 0.24 ml/kg (dose II) into the ear vein of 5 animals each.
Five rabbits served as control and received 0.24 ml/kg of a 0.9% NaCl-solution instead of GF.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

glycerol formal

IUPAC Name:

glycerol formal

Details on test material:

- Name of test material (as cited in study report): ST 2060
- Impurities (identity and concentrations): GF is stabilised with 0.02% ethylenediamine tetraacetic acid-sodium salt; 0.02% propylgallate and 0.01% thiodipropionic acid
- Mixture in constant ration of 5-hydroxy-1,3-dioxane (60%) and 4-hydoxymethyl-1,3-dioxolane (40%)

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Own breeding
- Age at study initiation: not specified
- Weight at study initiation: not specified
- Fasting period before study: not specified
- Housing: separate containers with a wire floor
- Diet (e.g. ad libitum): Altromin-K-pellets ad libitum
- Water (e.g. ad libitum): water ad libitum
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
not specified

Administration / exposure

Route of administration:

intravenous

Vehicle:

other: NaCl solution

Details on exposure:

GF was applied daily during 4 weeks as 30% dilution with 0.9% NaCl-solution

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

28 days

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:
Dose I group: 0.024 ml/kg
Dose II group: 0.24 ml/kg

No. of animals per sex per dose:

Control group: 4 female and 1 male
Dose I group (0.024 ml/kg): 3 female and 2 males
Dose II group (0.24 ml/kg): 3 female and 2 males

Control animals:

yes, concurrent vehicle

Details on study design:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: daily

HAEMATOLOGY: Yes
- Time schedule for collection of blood: before begin of treatment and afetr 1-, 2- and 4-weeks application of GF
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5 per group
- Parameters examined: erythrocyte, retyculocyte and leucocyte count, colorimetric determination of hemoglobin, determination of the hematocrit and differentiation of the white blood cells

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 1-and/or 4-weeks
- Animals fasted: No data
- How many animals: 5 per group
- Parameters examined: Sulfhydryl groups in erythrocytes (after 1- and 4-weeks); bilirubin in plasma (after 4-weeks); hemoglobin in plasma (afetr 1- and 4-weeks)

Sacrifice and pathology:

Animals were anesthetised with 5 ml Nembutal (intravenous route) and were sacrified by exsanguination after termination of the study.
Following organs were taken from animals and fixed in 10% formol after determination of their weight: liver, spleen, kidney, hearth, aorta, lung, adrenal glands and thyroid of all animals; ovary and uterus of females; testes and epididymes of males.

Statistics:

Statisics were performed with indication of statistical significances as against control

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not examined

Details on results:

HAEMATOLOGY and CLINICAL CHEMISTRY
Significant differences in coparison with control have been observed in:
- 0.24 ml/kg dose group after 4 weeks application for erythrocytes count (95%)
- 0.24 ml/kg dose group after 2 and 4 weeks application for hemoglobin % (95%)
- 0.24 ml/kg dose group after 4 weeks application for hematocrit % (99.9%)
- 0.024 ml/kg dose group after 2 weeks application for Mean Corpuscular Volume (MCV) (95%)
- 0.024 ml/kg dose group after 2 weeks application for Mean Corpuscular Hemoglobin (MCH) (95%)
- 0.24 ml/kg dose group after 4 weeks application for retyculocytes count (95%)
- 0.024 ml/kg dose group after 2 weeks application for leucocyte count (95%)
- 0.24 ml/kg dose group after 1 week application for leucocyte count (99%)

From these hematological data, a decrease of the erythrocytes, the hemoglobin and the hematocrit was recorded after intravenous application of GF at 0.24 ml/kg. This increase is to be attributed to the hemolytical potency of GF. A reduction of the non-protein bound SH-groups in the erythrocytes was observed without appearance of hemoglobin in the plasma.

GROSS PATHOLOGY and HISTOPATHOLOGY
Observations in control group:
The studies revealed the following findings deviating from the perfect structure:
- Liver: single round cell nodules in 1 animal
- Kidney: round cell nodules in 1 animal
- Aorta: calcareous degenerations in the media of 4 animals
- Uterus: inflammatory changes in 1 animal

Observations in dose I group (0.024 ml/kg):
The studies revealed the following findings deviating from the perfect structure:
- Liver: single round cell nodules in 2 animals
- Kidney: single round cell nodules in 1 animal and 1 edema of the cortica in another animal
- Aorta: calcereous degenerations of the media in 3 animals

Observations in dose II group (0.24 ml/kg):
The studies revealed the following findings deviating from the perfect structure:
- Liver: slight to moderate changes in all animals
- Kidney: sporadically round cell filtrates in 2 animals
- Aorta: changes in the wall in 2 animals
- Testis: reduced spermiogenesis in 1 animal

Differences in comparison to the organs of the control animals appeared only in the liver in dose II group. The changes are not grave.

Effect levels

Dose descriptor:

NOAEL

Remarks:

: The author stated that differences in comparison to the organs of the control animals appeared only in the liver in dose II group. The changes are not grave.

Remarks on result:

not determinable

Remarks:

no NOAEL identified

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

It can be concluded that sufficient safety is given for the application of stabilised GF as solubilizer for pharmaceutical forms of administration.

The author stated that differences in comparison to the organs of the control animals appeared only in the liver in dose II group. The changes are not grave.

Executive summary:

Toxicity tests after intravenous (i.v.) application of glycerol formal was performed with rabbits for a period of 4 weeks.

The test item was applied daily as 30% dilution with 0.9% NaCl-solution at dose of 0.024 ml/kg (dose I) and 0.24 ml/kg (dose II) into the ear vein of 5 animals each.

Five rabbits served as control and received 0.24 ml/kg of a 0.9% NaCl-solution instead of GF.

The following controls ans examinations were made during the test:

- Daily observation of the general behaviour and control of weight.

- Hematological studies (erythrocyte-, leucocyte- and reticulocyte count, hemoglobin, hematocrit, differential blood picture) before begin of the application and after 1, 2 and 4 weeks of application of GF.

- Determination of hemoglobin content in plasma and sulfhydryl groups in erythrocytes after 1 and 4 weeks of GF application. - - Determination of bilirubin in the plasma after 4 weeks of GF application

- Histo-pathological tests after 4 weeks of GF treatment.

No differences in comparison with the controls were observed for:

- the general behaviour and course of weight

- the bilirubin values

- the appearence and weight of the organs during the post-mortem examination

- the relative organ weight

The 0.24 ml/kg dose treatment caused a decrease of eruthrocytes, hemoglobin and hematocrit, yet no hemoglobin was detectable in the plasma. The erythrocytes showed a decrease of the SH-groups after 1 and after 4 weeks of treatment.

The histological examinations of liver revealed increased, enlarged cells charged with fat, whereas the testes showed symptoms of a reduced spermiogenesis in the dose II group. The other organs showed no deviating findings.

The 4 weeks intravenous application of the human maximum dose (Dose I: 0.024 ml/kg) adjusted to the body weight (70 kg) for stabilised glycerol formal caused in rabbits no findings deviating from those of the controls. The 10 times higher dosage (Dose II: 0.24 ml/kg) decreased the erythrocytes number, Hb-content and erythocytes content of SH-groups and showed in histological section signs of alteration of the tissue in liver and testes.

It can be concluded that sufficient safety is given for the application of stabilised GF as solubilizer for pharmaceutical forms of administration.

The author stated that differences in comparison to the organs of the control animals appeared only in the liver in dose II group. The changes are not grave.