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

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP lab following OECD guideline

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

glycerol formal

IUPAC Name:

glycerol formal

Test material form:

other: liquid

Details on test material:

. Name : Glycerol formal
. Synonym : Glycerol Formal Ultra Pure Grade
Both denominations correspond to the same test item.
. CAS number : 4740-78-7 + 5464-28-8 (ratio ~ 50/50)
. Batch number : 1206191600R
. Supplier : LAMBIOTTE & CIE S.A
. Description : colorless liquid
. Container : brown glass flask
. Purity/Composition : 99.9840%
. Storage condition : at room temperature
. Date of receipt : 27 July 2015
. Expiry date : 19 June 2015

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

Animals
Number: six nulliparous and non-pregnant female rats were received at CiToxLAB France on 09 and 14 August 2012.
Strain and Sanitary status: Sprague-Dawley rat, Rj Han: SD, Indemn of Organism Pathogen Specific Han (IOPS Han).
Breeder: Janvier, Le Genest-Saint-Isle, France.
Age/Weight: on the day of treatment, the females were approximately 8 weeks old and had a mean body weight of 215 g (range: 208 g to 220 g).
Receipt: on arrival, the animals were given a clinical examination to ensure that they were in good condition.
Acclimation: the animals were acclimated to the study conditions for a period of 5 days before treatment.
Allocation to groups: the day before treatment, the required number of animals (three females per treatment step) were allocated to the groups according to a computerized randomization procedure.
Identification: the day before treatment, the animals were individually identified by ear notches (unique CiToxLAB France identity number).

Environmental conditions
From arrival at CiToxLAB France, the animals were housed in a barriered rodent unit.
The animal room conditions were set as follows:
. temperature : 22 ± 2°C,
. relative humidity : 50 ± 20%,
. light/dark cycle : 12h/12h,
. ventilation : approximately 12 cycles/hour of filtered, non-recycled air.
The corresponding instrumentation and equipment are checked and calibrated at regular intervals. The temperature and relative humidity are recorded continuously (recording devices equipped with alarm systems).

Housing
The animals were housed by three from the same group in polycarbonate cages with stainless steel lids (Tecniplast 2154, 940 cm²) containing autoclaved sawdust (SICSA, Alfortville, France). Rat Nylabone was given as enrichment.

Food and water
All animals had free access to SSNIFF R/M-H pelleted maintenance diet, batch No. 2626975 (SSNIFF Spezialdiäten GmbH, Soest, Germany), and tap water (filtered with a 0.22 μm filter).
During periods of fasting, food was removed, but the animals were not deprived of water.

Contaminant analyses
The batches of diet and sawdust were analyzed by the suppliers for composition and contaminant levels. Bacterial and chemical analyses of water are performed regularly by external laboratories. These analyses include the detection of possible contaminants (pesticides and heavy metals). No contaminants that could have interfered with or prejudiced the outcome of the study were found in the diet, drinking water or sawdust.

Administration / exposure

Route of administration:

oral: drinking water

Vehicle:

water

Remarks:

The vehicle was drinking water treated by reverse osmosis using ELIX 5 (Millipore SA, Saint-Quentin en Yvelines, France).

Details on oral exposure:

Vehicle
The vehicle used in this study was selected from the results of solubility assays performed by the CiToxLAB France Pharmacy.
In the absence of recommendation from the Sponsor, the solubility assay first started at the concentration of 200 mg/mL, and the first choice vehicle was drinking water treated by reverse osmosis. A solution was obtained at the concentration of 200 mg/mL in this vehicle.

Dose formulation preparation
The test item was administered as a solution in the vehicle. The test item was mixed with the required quantity of vehicle.
Dose formulations preparations were prepared by the CiToxLAB France Pharmacy extemporaneously on the day of each administration.
The dose formulations were stored at room temperature and delivered to the study room in brown flasks.

Administration
The dose formulations were administered once by gavage, using a plastic syringe fitted with a plastic gavage tube.
The quantity of dose formulation administered to each animal was adjusted according to the body weight recorded on the day of dose administration. A constant dosage-volume of 10 mL/kg was used. The dose formulations were kept at room temperature and were stirred continuously throughout the dosing procedure.

Doses:

Rationale for dose-level selection
The starting dose-level was selected in agreement with the Sponsor, based on the following rationale:
. based on available test item toxicity data, no morbidity or mortality was expected to occur at the dose-level of 2000 mg/kg. This was therefore chosen as the starting dose-level.
After treatment at the starting dose-level, the next dose-levels were administered in a sequential manner, under the same conditions to different animals, based on the dose-levels indicated in the flow charts of OECD Guideline No. 423, 17th December 2001, which are equivalent to those of Commission Regulation (EC) No. 440/2008, B.1tris (30 May 2008).

No. of animals per sex per dose:

Three female animals were used at each dose-level. The time intervals between the treatment of each group were 7 days and were determined by the onset, duration and severity of any signs of toxicity. The next group was treated according to the results obtained for the previously treated group.

Control animals:

no

Details on study design:

Duration
The dose formulations were administered once followed by an observation period of 14 days. For each animal, the day of dose administration was recorded as day 1 of its observation period.

CLINICAL EXAMINATIONS

Morbidity and mortality
Each animal was checked for mortality and morbidity, frequently during the hours following administration, then at least once a day until the end of the observation period, including weekends and public holidays.

Clinical signs
Each animal was observed after treatment as follows:
. at least once during the first 30 minutes,
. periodically during the first 4 hours,
. then once a day, at approximately same time, for the recording of clinical signs.
Any clinical signs observed were recorded individually for each animal, along with the times of onset and recovery.

Body weight
The body weight of each animal was recorded the day of group allocation then on the day of treatment and on days 8 and 15.
The body weight gain of the test item-treated animals was compared to that of CiToxLAB France historical control data generated from animals of the same strain and age treated with drinking water treated by reverse osmosis under similar experimental conditions.

PATHOLOGY

Euthanasia
On completion of the observation period, animals were deeply anesthetized by an intraperitoneal injection of pentobarbital sodium and euthanized by exsanguination.

Macroscopic post-mortem examination
A macroscopic post-mortem examination was performed on all animals. After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed. All gross observations were recorded individually for each animal.

Preservation of tissues
For all animals, the macroscopic lesions were preserved in 10% buffered formalin. Histological specimens (tissues in fixative) were destroyed at the finalization of the study report.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No unscheduled deaths occurred during the study.

Clinical signs:

other: No clinical signs were observed in any animals.

Gross pathology:

There were no macroscopic post-mortem findings related to test item treatment.
The macroscopic findings observed correlated with common histological findings. Thus they were
considered to be incidental.

Any other information on results incl. tables

Table 1: Body weight (BW) changes in g

	CiToxLAB France historical control data	Group 1 (2000 mg/kg/bw)	Group 2  (2000 mg/kg/bw)
BW (Day 1)	219	213	216
BW (Day 8)	264	244	255
BW (Day 15)	284	263	271
BW change (Days 1 -8)	+44	+31	+39
BW change (Days 8 -15)	+20	+19	+15
BW change (Days 1 -15)	+64	+50	+54

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions of this study, the oral LD50 of the test item, Glycerol formal, was higher than 2000 mg/kg in rats.
Therefore, the test item is not classified as toxic by oral route according to the criteria of CLP Regulation.

Executive summary:

The objective of this study was to evaluate the potential acute toxicity of the test item, Glycerol formal, following a single oral administration (gavage) to rats.

Methods

The test item, Glycerol formal (batch No. 1206191600R), was administered once by oral route (gavage) to two groups of three fasted female Sprague-Dawley rats under a dosage-volume of 10 mL/kg. The test item was prepared in drinking water treated by reverse osmosis. Based on available test item toxicity data, the starting dose-level of 2000 mg/kg was chosen. After the first assay, as no toxicity was observed, the results were confirmed in other females. Each animal was observed at least once a day for mortality and clinical signs for 15 days. Body weight was recorded on day 1 and then on days 8 and 15. On completion of the observation period, the animals were sacrificed and then submitted for a macroscopic post-mortem examination. Macroscopic lesions were preserved in buffered formalin then destroyed at the finalization of the study report as no microscopic examination was performed.

Results

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals. When compared to CiToxLAB France historical control data, a trend to a slightly lower mean body weight gain was observed in 3/6 females during the first week of study and 2/6 females during the last week of the study. There were no macroscopic post-mortem findings related to test item treatment.

Conclusion

Under the experimental conditions of this study, the oral LD50 of the test item, Glycerol formal, was higher than 2000 mg/kg in rats. Therefore, the test item is not classified as toxic by oral route according to the criteria of CLP Regulation.