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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP lab following OECD guidelines

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 402 (Acute Dermal Toxicity)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes

Test type:

standard acute 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
. Other name : Glycerol Formal Ultra Pure Grade
Both names refer to the same test item. The name retained in the study report is Glycerol formal.
. CAS number : 4740-78-7 and 5464-28-8 (~ 50/50 ratio)
. Batch number : 1206191600R
. Supplier : LAMBIOTTE & CIE S.A.
. Description : colourless liquid
. Container : brown glass flask
. Purity : 99.9840%
. Storage condition : at room temperature
. Date of receipt : 12 July 2012
. Expiry date : 19 June 2015

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

ANIMALS
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 animals were approximately 8 weeks old. The males had a mean body weight of 376 g (range: 370 g to 387 g) and the females had a mean body weight of 228 g (range: 218 g to 242 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 7 days (females) or 10 days (males) before treatment.
Allocation to study: the day before treatment, the required number of animals was allocated by sex to the groups according to a computerized randomization procedure. The skin of each animal (previously clipped) was examined in order to use only animals with healthy intact skin.
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 (see § Study plan adherence),
. 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 and the records checked daily and filed.

Housing
The animals were housed by five from the same sex and group in polycarbonate cages with stainless steel lids (Tecniplast 2000P, 2065 cm2) containing autoclaved sawdust (SICSA, Alfortville, France). 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).

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

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Preparation of the animals
On the day before treatment, an area representing approximately 10% of the total body surface area (i.e. approximately 7 cm x 5 cm for males and 6 cm x 5 cm for females (1)) was clipped on the back of each animal using an electric clipper. Care was taken to avoid abrading the skin. Only animals with healthy intact skin were used for the study.

(1) The total body surface is calculated according to Meeh's formula: S = k.p2/3 where S = body surface (cm2), p = body weight (g), k = 7.47 for rat [Diack, S L. (1930). The determination of the surface area of the white rat. J. Nutr. 3(3): 289-296].

Duration of exposure:

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

Doses:

A limit test was carried out by application of 2000 mg/kg to five females in the first instance.
As no mortality occurs, the limit test was completed using five males. As no mortality also occurs in males, the study was considered as complete.

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

Application
The test item was applied as a film (as thin and uniform as possible) to the clipped area of skin. The application site was covered with a hydrophilic gauze pad.
The quantity of test item applied to each animal was adjusted according to the body weight recorded on the day of dose application.
The gauze pad was held in place with an aerated hypoallergic dressing for 24 hours to avoid ingestion of the test item by the animals.
No residual test item was noted after removal of the dressing.

CLINICAL EXAMINATIONS
Morbidity and mortality
Each animal was checked for mortality and morbidity, frequently during the hours following treatment, then 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.
From day 2, any local reactions at the treatment site were recorded.

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.

Statistics:

not necessary as a limit test was performed.

Results and discussion

Effect levels

Sex:

male/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 indicative of systemic toxicity were observed in any animals. A well-defined erythema was noted at the application site on day 2 and then a very slight erythema associated with a very slight dryness was observed from day 3 to day 6 in 1/

Gross pathology:

There were no macroscopic changes at necropsy.

Any other information on results incl. tables

Table 1 - body weight evolution

	CiToxLAB France historical control data (FEMALE)	Females exposed at 2000 mg/kg	CiToxLAB France historical control data (MALE)	Males exposed at 2000 mg/kg
BW (day 1)	228	228	332	376
BW (day 8)	264	246	377	418
BW (day 15)	283	265	422	419
BW change (days 1 - 8)	+36	+18	+45	+41
BW change (days 8 - 15)	+18	+19	+45	+2
BW change (days 8 - 15)	+55	+37	+90	+43

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

higher than 2000 mg/kg in rats.
Therefore, the test item should not be classified as toxic by dermal route according to the criteria of CLP
Regulation.

Executive summary:

The objective of this study was to evaluate the potential toxicity of the test item following a single dermala pplication to rats.

Methods

The test item, Glycerol formal, was applied in its original form to the skin of five female then five male Sprague-Dawley rats at the dose-level of 2000 mg/kg. The application site was covered by a semi-occlusive dressing for 24 hours.

Each animal was observed at least once a day for mortality and clinical signs for 15 days. From day 2, any local reactions at the treatment site were also noted. 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. No tissues were preserved.

Results

No unscheduled deaths occurred during the study and no clinical signs indicative of systemic toxicity were observed in any animals.

A well-defined erythema was noted at the application site in 1/5 females on day 2 then a very slight erythema associated with a very slight dryness was observed from day 3 to day 6.

No cutaneous reactions were observed in any males during the study period.

When compared to CiToxLAB France historical control data, lower mean body weight and mean body weight gain were recorded in both sexes during the study.

No macroscopic changes were observed at necropsy.

Conclusion

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