Reaction products of 2-(chloromethyl)oxirane and glycerol

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: well documented GLP-guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Remarks:

- only minor deviation (relative humidity sometimes outside of the target ranges specified in the protocol). This minor deviation was not considered to comprise the validity or integrity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Remarks:

- only minor deviation (relative humidity sometimes outside of the target ranges specified in the protocol). This minor deviation was not considered to comprise the validity or integrity of the study.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of 2-(chloromethyl)oxirane and glycerol

EC Number:

701-197-2

Molecular formula:

6 individual glycidether components (Molecular range ca. 200 - 600 g/mol)

IUPAC Name:

Reaction products of 2-(chloromethyl)oxirane and glycerol

Test material form:

other: liquid

Details on test material:

- Name of test material: 1,2,3-Propanetriol, glycidyl ethers
- Substance type: organic
- Physical state: liquid
- Storage condition of test material: at room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS - Sprague-Dawley ICO: OFA-SD (IOPS Caw)
- Source: Iffa Credo, L'Arbresle, France
- Age at study initiation: on the day of treatment, the animals were approximately 6 weeks old
- Weight at study initiation: mean body weight +/- standard deviation of 191 +/- 6 g for the males and 144 +/- 7 g for the females.
- Fasting period before study: overnight (approximately 18 hours, but had free access to water. Food was given back approximately 4 hours after administration of the test substance
- Housing: The animals were housed in polycarbonate cages (48 cm * 27 cm * 20 cm). Each cage contained four to seven animals of the same sex during the acclimatization period and five rats of the same sex during the treatment period. Each cage contained dust-free sawdust. Bacterial and chemical analyses of the sawdust, including the detection of possible contaminants (pesticides, heavy metals), are performed regularly by external laboratories.
- Diet (e.g. ad libitum): All animals had free access to A04C pelleted diet (UAR, Villemoisson-sur-Orge, France), except as noted above for fasting purposes. Each batch of food was analysed by the supplier for composition and contaminant levels.
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided at libitum. Bacteriological and chemical analyses of the water and diet, including the detection of possible contaminantes (pesticided, heavy metals and nitrosamines), are performed regularly by external laboratories.
No contaminants are known to be present in the diet, drinking water or bedding material at levels which may be expected to interfere with or prejudice the outcome of the study.
- Acclimation period: at least 5 days
- Identification. the animals were identified individually by earmarks of earmotches

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 2°C
- Humidity (%): 30 - 70 %
- Air changes (per hr): approximately 12 cycles/hours of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12 h/ 12 h

The temperature and relative humidity were under continuous control and recording. The records were checked daily and retained. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular intervals.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: unchanged for 2000 mg/kg bw, corn oil in the dose-levels 1415 mg/kg and 1000 mg/kg

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 141,5 mg/mL or 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: commonly used and well established
- Lot/batch no. (if required): 86H0059

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

Doses:

2000 (undiluted), 1415 (diluted in corn oil), and 1000 mg/kg (diluted in corn oil)

No. of animals per sex per dose:

2000mg/kg dose-level: 5 males and females
1415mg/kg dose-level: 5 females
1000mg7Kg dose-level: 5 females

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
The animals were observed frequently during the hours following administration of the test substance, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until day 15. Type, time of onset and duration of clinical signs were recorded for each animal individually. Time of death was recorded individually, in terms of the number of hours or days after dosing.
Body weight - the animals were weighed individually just before administration of the test substance on day 1 and then on days 8 to 15. Individual weights of animals found dead during the study were measured at necropsy when survival exceeded 24 hours and if no signs of "cannibalism" were present. The body weight gain of the treated animals was compared to that of CIT control animals wi9th the same initial body weight.
- Necropsy of survivors performed: yes
The animals found dead during the study were subjected to a macroscopic examination as soon as possible. On day 15, all surviving animals were killed by carbon dioxide asphyxiation and a macroscopic examination was performed.
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. In case of macroscopic lesions, organ samples were taken and preserved in 10 % buffered formalin. No microscopic examination was performed.
- Other examinations performed: clinical signs, body weight,organ weights

Statistics:

Evaluation of the toxicity of the test substance following a single oral administration in rats should include the relationship, in any, between the animals' exposure to the test substance and the incidence and severity of all abnormalities including behavioural and clinical abnormalities, macroscopic lesions, body weight changes,mortality and any other toxic effects.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

no animals died in the 1000 and 1415 mg/kg dose groups. 3 of 5 males and 2 of 5 females died in the 2000 mg/kg dose group.

Clinical signs:

other: At the 2000 mg/kg dose-level, hypoactivity or sedtion and piloerection were observed in all animals from day 1 up to day 3 at the latest; coma was also observed prior to death in one female. Recovery was complete in the surviving animals on day 2. At the

Gross pathology:

macroscopic examinations of the main organs of the animlas revealed no apparent abnormalities

Any other information on results incl. tables

Table 1:Individual clinical signs and mortality
Dose	Time	Animals		Mortality	Clinical signs
(mg/kg)		Males	Females
1000	30 min		02-03	No	Hypoactivity
			01-04 -05	No	None
	1h-2h-4h		01-02-03-04-05	No	Hypoactivity
	D2 to D 15		01-02-03-04-05	No	None
1415	30 min - 1 h		01-02-03-04-05	No	None
	2h		01-02-05	No	Hypoactivity, piloerection
			03-04	No	None
	4h		01-02-03-04-05	No	None
	D 2 to D 15		01-02-03-04-05	No	None
2000	30 min - 1 h	01-02-03-04-05	01-02-03-04-05	No	None
	2h	01-02-03-04-05	01-02-03-04-05	No	Sedation, piloerection
	4h	02		Yes
			01	No	Piloerection, coma
		01-03-04-05	02-03-04-05	No	Hypoactivity, piloerection
	6h		01	No	Piloerection, coma
		01-03-04-05	02-03-04-05	No	Hypoactivity, piloerection
	D2 (morning)		01	Yes
		03-04		No	Hypoactivity, piloerection
		01 -05	02-03-04-05	No	None
	D2 (afternoon)	03-04		No	Hypoactivity, piloerection
		01-05	02-03-04-05	No	None
	D3 (morning)	03	03	Yes
		04		No	Hypoactivity
		01-05	02-04-05	No	None
	D3 (afternoon)	04		No	Hypoactivity
		01-05	02-04-05	No	None
	D4 (morning)	04		Yes
		01-05	02-04-05	No	None
	D > 4 (afternoon) to D 15	01-05	02-04-05	No	None
min:minutes
h  :hour
D  :day

Table 2: Individual and mean body weight and weekly body weight change of treated rats (g)
Dose	Volume	Sex	Animals	Days
mg/kg	ml/kg			1	(1)	8	(1)	15	At death
1000	10	Female	01	131	38	169	21	190	-
			02	136	35	171	23	194	-
			03	149	48	197	41	238	-
			04	136	42	178	43	221	-
			05	141	29	170	22	192	-
			M	139	38	177	30	207	-
			SD	7	7	12	11	21	-
1415	10	Female	01	152	36	188	22	210	-
			02	147	45	192	22	214	-
			03	149	50	199	26	225	-
			04	146	39	185	28	213	-
			05	147	39	186	24	210	-
			M	148	42	190	24	214	-
			SD	2	6	6	3	6	-
2000	1.67	Male	01	183	59	242	66	308	-
			02	188					-
			03	192					174
			04	200					-
			05	194	72	266	60	326	-
			M	191	66	254	63	317
			SD	6	9	17	4	13
2000	1.67	Female	0!	156					-
			02	149	42	191	27	218	-
			03	148					147
			04	140	35	175	21	196	-
			05	140	31	171	25	196	-
			M	347	36	179	24	203
			SD	7	6	11	3	13
(1)  -Body weight gain
M   -Mean
SD  -Standard Deviation
- = not applicable
Animals found dead during the study not mentioned

Table 3: Individual macroscopic examinations at necropsy
Dose	Time	Animals		Macroscopic abnormalities
mg/kg		Males	Females
1000	D15		01-02-03-04-05	None
1415	D15		01-02-03-04-05	None
2000	D1	02		No apparent abnormalities
	D2		01	Advanced autolysis
	D3	03	03	Advanced autolysis
	D4	04		Cannibalized animal
	D15	01-05	02-04-05	No apparent abnormalities
D: day

Applicant's summary and conclusion

Interpretation of results:

other: Category 4 based on EU GHS criteria

Remarks:

Criteria used for interpretation of results: other: EU-GHS

Conclusions:

LD50 is near 2000 mg/kg bw. C&L is required.

Executive summary:

The test substance was administered via oral gavage to groups of five male and/or female fasted Sprague-Dawley rats. In the first instance, the test substance was administered undiluted, at the dose of 2000 mg/kg (dose volume 1.67 ml/kg, taking into consideration that its specific gravity was 1.2. As 50 % mortality occurred in this limit test, the test substance was then prepared in corn oil and administered to other animals at lower doses (1415 mg/kg and 1000 mg/kg in corn oil (dose volume 10 ml/kg) to 5 females each). Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test substance. All animals were subjected to necropsy. At the 2000 mg/kg dose-level 3/5 males and 2/5 females died between days 1 and 4. Hypoactivity or sedation and piloerection were observed in all animals from day 1 up to day 3 at the latest, coma was observed prior to death in one female. Recovery was complete in the surviving animals on day 2. At the 1415 mg/kg dose-level, no mortality was observed. Hypoactivity and piloerection were noted in 3/5 females on day 1. At the 1000 mg/kg dose-level, no mortality was observed. Hypoactivity was noted in all females on day 1. The body weight gain of the surviving animals was not affected by treatment with the test substance. No apparent abnormalities were observed in all the animals at necropsy. In conclusion and under the experimental conditions, the oral LD50 of the test substance is near 2000 mg/kg. In accordance with ethic and scientific recommendations concerning the LD50, a more precise determination was not conducted. According to the classification criteria laid down in Commission Directive 93/21/EEC, concerning the potential toxicity by the oral route, the test substance should be assigned the symbol Xn, the indication of danger "Harmful" an the risk phrase R 22 " Harmful if swallowed".