Ethyl 3,5-dichloro-4-hexadecyloxycarbonyloxybenzoate

Endpoint summary

Description of key information

Oral (OECD 401), rat: LD50 > 5000 mg/kg bw (limit test)
Dermal (OECD 402), rat: LD50 > 2000 mg/kg bw (limit test)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from November 28th, 1988 to December 12th, 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study performed according to OECD Guidelines for testing of chemicals (no 401)

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Wistar outbred (Bor:WISW)

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animals
Mult, about 16 weeks old, albino rats were used. They ware Wistar outbred
rats (Bor:WISW) obtained from a colony, maintained under SPF—conditions at
the Breeding Centre for Laboratory Animals, F. Winkelmann GmbH, Borchen,
F.R. Gerinany. The body weights of the males varied from 308 to 354 g, those
of the females from 182 to 202 g. The animals ware kept under the
environmental conditions of the Institute’s animal house for about 13 weeks
prior to the test.
Housing and maintenance
The rats ware housed in groups of f ive, males and females separated, in
stainless steel cages with wire—screen bottom and front, in a room
ventilated with about 10 air changes per hour and maintained at 20—24°C.
Relative humidity was regulated between 40 and 70 per cent, lighting was
artificial by fluorescent tubes, time switch—controlled at a sequence of
12 hours light, 12 hours dark. Tap water was freely available at all times
by means of an automatic watering system. The rats had free access to the
Institute’s cereal—based, open—formula diet for rats and mice,
except in the overnight period before dosing tili 4 hours after dosing,
when food was withheld. The diet is analyzed regularly for nutrients and
contaminants. Tap water is analyzed regularly for
contaminants.

Route of administration:

oral: gavage

Vehicle:

maize oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25%
- Amount of vehicle (if gavage): 20 mL/kg bw
- Justification for choice of vehicle: maize oil was selected as it was shown to be an appropriate vehicle in a pre-test study

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg bw

Doses:

one single dose of 20.0 ml per kg body weight
This dose level is equal to 5000mg test substance per kg body weight

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Preliminary study:

A preliminary study was carried out to find an appropriate vehicle and the
general level of acute toxicity of the test substance.
After these preliminary observations, the substance was given by gavage as
a 25% (w/v) suspension in maize oil, in one single dose of 20.0 ml per kg
body to 5 males and 5 females. This dose level is equal to 5000 mg test
substance per kg body weight. 1f a dose level of 5000 mg/kg or more does
not cause compound—related mortality, then a full study with various dose
levels is not required (OECD—Guidelines for Testing of Chemicals).
The rats were observed frequently for signs of intoxication, during the
first 4 hours after treatment and thereafter, at least once daily
throughout an observation period of 14 days. The individual body weights of
the rats were recorded on day 0, 3, 7 and 14. At the end of the observation
period, the rats were killed and examined grossly.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

>= 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured and all rats looked quite heathly throughout the remaining part of the 14-day observation period

Clinical signs:

other: Except signs of sluggishness and piloerection during the first few hours after treatment no signs of intoxication were observed

Gross pathology:

Macroscopic examination of the rats at the termination of the study
revealed a mottled surface of one of the kidneys in one male and one
female. No other treatment—related gross alterations were found.
At necropsy, 1/5 males and 1/5 females had a mottled surface of one kidney, which is not considered to be substance-related. No other gross abnormalities were observed.

Table 1: Mortality and clinical signs

Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death	Mortality (%)
Males
5000	0/5/5	1-24 h	-	0
Females
5000	0/5/5	1-24 h	-	0
Overall LD50 > 5000 mg/kg bw

* first number = number of dead animals                                 

 second number = number of animals with systemic clinical signs         

 third number = number of animals used                               

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

From the mortality figures, the oral LD50 of “AF—366” was found to exceed
5000 mg per kg body weight, both for male and for female rats. Therefore,
the test substance can be classified as unharmful (EC—directive 83/467/EEC,
Official Journal of the European Communities, L 257, September 16, 1983).
Not classified.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 Nov-14 Dec 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. The test substance (a solid) was applied under occlusive conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted in February 1987

Deviations:

yes

Remarks:

exposure under occlusive conditions

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Wistar outbred (Bor:WISW)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding Centre for Laboratory Animals, F. Winkelmann GmbH, Borchen, Germany
- Age at study initiation: approximately 19 weeks
- Weight at study initiation: 310-360 g (males), 190-230 g (females)
- Housing: before dosing, the rats were housed in groups of five per sex, in stainless steel cages with a wire-screen bottom and front. After dosing, the rats were housed individually in stainless steel cages with a wire-screen bottom and front.
- Diet: CIVO cereal-based, open-formula diet for rats, ad libitum
- Water: tap water, ad libitum
- Acclimation period: approximately 14 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24
- Humidity (%): 40-60
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 30 Nov 1988 To: 14 Dec 1988

Type of coverage:

occlusive

Vehicle:

maize oil

Details on dermal exposure:

TEST SITE
- Area of exposure: the dorsal and ventral clipped area of the trunk of the animals
- Type of wrap if used: the test material was held in contact with the skin with a plastic film, which was secured by an elastic adhesive bandage (Tensoplast) around the trunk of the rat

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the residual test material was cleaned from the treated area with water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 8 mL/kg bw
- For solids, paste formed: no, a suspension was prepared

VEHICLE
- Amount(s) applied (volume or weight with unit): 8 mL/kg bw
- Concentration (if solution): 75%

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations of clinical signs were made several times on Day 0 (post-administration) until 4 hours after dosing and daily thereafter; weighing was performed on Day 0 (pre-administration), 3, 7 and 14; skin irritation effects were assessed on Day 1, 3 and 7
- Necropsy of survivors performed: yes, all gross pathological changes were recorded
- Other examinations performed: clinical signs, mortality, skin irritation

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There was no mortality during the 14-day study period (see Table 1).

Clinical signs:

other: 5/5 males and 5/5 females were moderately sluggish from 1 up to 4 hours after administration of the test substance. Piloerection was observed at the 24-hour reading time point only, in 5/5 males and 5/5 females (see Table 1).

Gross pathology:

No treatment-related effects were observed during the gross pathological examination.

Other findings:

- Other observations: there were no skin irritation effects following exposure to the test substance

Table 1: Mortality and clinical signs

Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death	Mortality (%)
Males
2000	0/5/5	1-24 h	-	0
Females
2000	0/5/5	1-24 h	-	0
Overall LD50 > 2000 mg/kg bw

* first number = number of dead animals                                 

 second number = number of animals with systemic clinical signs         

 third number = number of animals used                               

 

Table 2: Individual and mean body weight changes

Body weight (g)
	Day 0	Day 3	Day 7	Day 14
Males
1	344	333	346	354
2	310	304	320	328
3	350	342	363	375
4	358	352	354	354
5	360	360	361	372
Mean body weight, males (g)	344	338	349	357
6	228	223	223	220
7	190	180	190	180
8	230	221	204	221
9	203	200	208	201
10	196	193	193	196
Mean body weight, females (g)	209	203	204	204

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified
DSD: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC)No 1907/2006

Additional information

The acute toxicity of ethyl 3,5-dichloro-4- hexadecyloxycarbonyloxybenzoate (AF-366) was assessed in an oral toxicity study and a dermal toxicity study.

In an acute oral toxicity study performed according to OECD guideline 401, the limit dose of 5000 mg/kg of the test substance was administered to 5 rats/sex by gavage (Spanjers, 1988a). No mortality was observed, and there were no effects on body weight during the 14-day observation period. The necropsy and gross pathological examination did not reveal any treatment-related effects. Therefore, the LD50 is > 5000 mg/kg bw for male and female rats.

The acute dermal toxicity of ethyl 3,5 -dichloro-4 -hexadecyloxycarbonyloxybenzoate was assessed in a limit test that was performed according to OECD 402 (Spanjers, 1998b). A single dose of 2000 mg/kg bw of the test substance was applied as a 25% solution in maize oil to the shaved skin of rats under occlusive conditions and left for 24 hours. There was no mortality during the 14-day observation period. The mean body weight of the females was reduced from day 3 until sacrifice on day 14. The necropsy and gross pathological examination did not show any treatment-related effects. Therefore, the LD50 (dermal) is > 2000 mg/kg bw for male and female rats.

Justification for classification or non-classification

The available data on acute toxicity of the test substance does not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and is therefore conclusive but not sufficient for classification.