Reaction products of 1H-Imidazole-1-ethanol, 4,5-dihydro-, 2-(C7-C17 odd-numbered, C17-unsatd. alkyl) derivs. and sodium hydroxide and chloroacetic acid

Endpoint summary

Description of key information

The endpoint conclusion for acute oral toxicity is based on three key studies (reliability 2) and several supporting studies. The acute dermal toxicity has been investigated in rats by a limit test: LD50 dermal: > 2612 mg/kg

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study meets the EC Standards (conducted equivalent to OECD 401), although prior to GLP principles.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

lack of reported data; no body weight and no necropsy findings

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: OLAC 1976, Shaws Farm, Blackthorn, Bicester
- Age at study initiation: young adults
- Weight at study initiation: 160-250 g
- Fasting period before study: overnight before dosing
- Housing: in groups of five rats of one sex in polythene breeding cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21 degrees Celsius
- Humidity (%): 50-70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 10/14

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DOSE VOLUME APPLIED:
Pilot study: 0.85 - 7.6 mL
Main study: 6.0 - 12.0 mL

Doses:

Pilot study: 40, 20, 10 and 5 ml/kg bw
Main study: 60, 42.4 and 30 ml/kg bw

No. of animals per sex per dose:

Pilot study: 1 female and 1 male rat per dose
Main study: 5 female and 5 male rats per dose

Control animals:

other: not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: immediately after dosing, 4 hours after dosing and then daily for 14 days
- Frequency of weighing: only immediately before dosing
- Necropsy of survivors performed: no

Statistics:

The data obtained was subjected to statistical evaluation using the method of Litchfield and Wilcoxon (J. Pharm. Exp. Therap. 1949, 95, 99)

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

42 mL/kg bw

Based on:

test mat.

Remarks:

(aqueous solution)

95% CL:

>= 33 - <= 53

Mortality:

Pilot study:
20, 10 and 5 ml/kg: none of the rats died
40 ml/kg: the female rat dosed at this level died 24 hours after dosing
Main study:
60 ml/kg: one male and 2 female rats died within 4 hours of dosing. The remaining rats died within 24 hours of dosing.
42.4 ml/kg: two female rats died within 24 hours and 1 male rat died within 48 hours.
30 ml/kg: two of the male rats died within 24 hours

Clinical signs:

other: lethargy, hunched stance

Gross pathology:

no data

Other findings:

no data

Dose level; aqueous solution (mL/kg)	No. of rats	Sex (M or F)	Average Bodyweight (g); immediately before dosing	Volume dosed (mL)	Mortality
					0-4 hrs.	1-7 days	8-14 days	Total
40.0	1	M	190	7.6	0	0	0	0	1/2
	1	F	180	7.2	0	1	-	1
20.0	1	M	180	3.6	0	0	0	0	0/2
	1	F	170	3.4	0	0	0	0
10.0	1	M	170	1.7	0	0	0	0	0/2
	1	F	170	1.7	0	0	0	0
5.0	1	M	170	0.85	0	0	0	0	0/2
	1	F	175	0.88	0	0	0	0
60.0	5	M	200	12	1	4	-	5	10/10
	5	F	190	11.4	2	3	-	5
42.4	5	M	220	9.3	0	1	0	1	3/10
	5	F	200	8.5	0	2	0	2
30.0	5	M	250	7.5	0	2	0	2	2/10

Pilot study:

20, 10 and 5 ml/kg: none of the rats died or showed any overt signs of toxicity throughout the 14 -day test period.

40 ml/kg: the female rat dosed at this level became lethargic 4 hours after dosing and died 24 hours after dosing. The male rat was lethargic for 48 hours after dosing, but showed no further overt signs of toxicity

Main study:

60 ml/kg: one male and 2 female rats died within 4 hours of dosing. The remaining rats were lethargic at the 4 hour observation period and died within 24 hours of dosing.

42.4 ml/kg: all the rats became lethargic and adopted a hunched stance within 4 hours of dosing. Two female rats died within 24 hours and 1 male rat died within 48 hours.

30 ml/kg: 3 male rats became lethargic within 24 hours. Two of these 3 rats died 24 hours later.

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 oral (rat): 42 ml/kg (aqueous solution)

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study meets the EC Standards (conducted equivalent to OECD 401), although prior to GLP principles.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

lack of reported data; no body weight and no necropsy findings

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: OLAC 1976, Shaws Farm, Blackthorn, Bicester
- Age at study initiation: young adults
- Weight at study initiation: 150 - 260 grams
- Fasting period before study: the animals were fasted overnight before dosing
- Housing: the animals were housed in groups of 5 rats of one sex in polythene breeding cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21 degrees Celsius
- Humidity (%): 50 - 70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 14 hours artificial light/day

IN-LIFE DATES: No data

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

VEHICLE: not applicable

Doses:

Pilot study: 5, 10, 20 and 40 ml/kg
Main study: 30, 42.4 and 60 ml/kg

No. of animals per sex per dose:

38 rats in total (19 male and 19 female);
Pilot study: 1 male and 1 female per dose
Main study: 5 male and 5 female per dose

Control animals:

other: not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: the animals were examined for overt signs of toxicity immediately after dosing, 4 hours after dosing and then daily for 14 days
- Necropsy of survivors performed: no
- Other examinations performed: no

Statistics:

The data obtained was subjected to statistical evaluation using the method of Litchfield and Wilcoxon (J. Pharm. Exp. Therap. 1949, 95, 99)

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

44.5 mL/kg bw

Based on:

test mat.

Remarks:

(aqueous solution)

95% CL:

>= 37 - <= 53

Mortality:

40; 20; 10 and 5 ml/kg bw: 0/2
30 ml/kg bw: 1/10
42.4 ml/kg bw: 3/10
60 ml/kg bw: 10/10

Clinical signs:

other: Starting from the dose level 30 mL/kg bw, rats were lethargic. Starting from the dose level 42.4 ml/kg bw, rats adopted a hunched stance.

Gross pathology:

no data

Other findings:

no data

Dose level; aqueous solution(mL/kg)	No. of rats	Sex (M or F)	Average Bodyweight (g); immediately before dosing	Volume dosed (mL)	Mortality
					0-4 hrs.	1-7 days	8-14 days	Total
40.0	1	M	170	6.8	0	0	0	0	0/2
	1	F	165	6.6	0	0	0	0
20.0	1	M	180	3.6	0	0	0	0	0/2
	1	F	150	3.0	0	0	0	0
10.0	1	M	160	1.6	0	0	0	0	0/2
	1	F	165	1.7	0	0	0	0
5.0	1	M	175	0.88	0	0	0	0	0/2
	1	F	160	0.80	0	0	0	0
60.0	5	M	170	10.2	0	5	-	5	10/10
	5	F	180	10.8	1	4	-	5
42.4	5	M	240	10.2	0	1	0	1	3/10
	5	F	190	8.1	0	2	0	2
30.0	5	M	250	7.5	0	1	0	1	1/10
	5	F	205	6.2	0	0	0	0

Pilot study:

No deaths and no overt signs of toxicity throughout the 14 day test period.

Main study:

- 60 ml/kg dose group: One female died within 4 hours, all the others were lethargic at the 4 hour observation period and died within 24 hours of dosing;

- 42.4 ml/kg dose group: all the rats were lethargic and adopted a hunched stance for 24 hours after dosing. One female died 24 hours after dosing and a further male and female rat had died by day 3. No further overt signs of toxicity throughout the 14 day test period;

- 1/10 death in the 30 ml/kg dose group (within 24 hours). One further male and two female rats were lethargic for 24 hours following dosing. No further overt signs of toxicity throughout the 14 day test period.

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 oral (rat): 44.5 ml/kg (aqueous solution)

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study meets the EC Standards (conducted equivalent to OECD 401), although prior to GLP principles.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

lack of reported data; no body weight and no necropsy findings

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: OLAC 1976, Shaws Farm, Blackthorn, Bicester, UK
- Age at study initiation: young adults
- Weight at study initiation: 160-250 g
- Fasting period before study: overnight before dosing
- Housing: in groups of five rats of one sex in polythene breeding cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21 degrees Celsius
- Humidity (%): 50-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 10/14

IN-LIFE DATES: no data

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

VEHICLE: Not applicable
DOSE VOLUME APPLIED: 0.85 - 10.8

Doses:

The dose levels used in the pilot study were 40, 20, 10 and 5 ml/kg.
The main study groups were dosed at 60, 42.4 and 30 ml/kg.

No. of animals per sex per dose:

Pilot study: 1 male and 1 female per dose level
Main study: 5 male and 5 female per dose level

Control animals:

other: not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: immediately after dosing, 4 hours after dosing and then daily for 14 days
- Frequency of weighing: not applicable
- Necropsy of survivors performed: no
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: no

Statistics:

The data obtained was subjected to statistical evaluation using the method of Litchfield and Wilcoxon (J. Pharm. Exp. Therap. 1949, 95, 99)

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

34 mL/kg bw

Based on:

test mat.

Remarks:

(aqueous solution)

95% CL:

>= 28 - <= 42

Mortality:

20, 10 and 5 ml/kg: none of the rats dosed at these levels died
40 ml/kg: the male rat dosed at this level died within 24 hours of dosing.
60 ml/kg: all rats died within 24 hours of dosing.
42.4 ml/kg: all five female rats died within 24 hours of dosing. One male rat died 24 hours after dosing and a further rat died 48 hours after dosing.
30 ml/kg: one male and one female rat died within 24 hours of dosing and a further male and female rat died 48 hours after dosing.

Clinical signs:

other: Starting from the dose level 30 mL/kg, the rats adopted a hunched stance and showed signs of lethargy

Gross pathology:

no data

Other findings:

no data

Dose level; aqueous solution(mL/kg)	No. of rats	Sex (M or F)	Average Bodyweight (g); immediately before dosing	Volume dosed (mL)	Mortality
					0-4 hrs.	1-7 days	8-14 days	Total
40.0	1	M	190	7.6	0	1	-	1	1/2
	1	F	165	6.6	0	0	0	0
20.0	1	M	175	3.5	0	0	0	0	0/2
	1	F	175	3.5	0	0	0	0
10.0	1	M	180	1.8	0	0	0	0	0/2
	1	F	165	1.8	0	0	0	0
5.0	1	M	175	0.88	0	0	0	0	0/2
	1	F	170	0.85	0	0	0	0
60.0	5	M	170	10.2	0	5	-	5	10/10
	5	F	180	10.8	0	5	-	5
42.4	5	M	245	10.4	0	2	0	2	7/10
	5	F	200	8.5	0	5	-	5
30.0	5	M	210	6.3	0	2	0	2	4/10
	5	F	200	6.0	0	2	0	2

Pilot study:

20, 10 and 5 ml/kg:

None of the rats dosed at these levels died or showed overt signs of toxicity throughout the 14 day test period

40 ml/kg:

The male rat dosed at this level became lethargic within 4 hours of dosing and died within 24 hours of dosing. The female rat showed no overt signs of toxicity.

Main study:

60 ml/kg: all rats became lethargic within 4 hours of dosing and died within 24 hours of dosing.

42.4 ml/kg: all five female rats died within 24 hours of dosing. One male rat died 24 hours after dosing and a further rat died 48 hours after dosing. The surviving rats became lethargic and adopted a hunched stance within 4 hours of dosing until the 48 hour observation period.

30 ml/kg: one male and one female rat died within 24 hours of dosing and a further male and female rat died 48 hours after dosing. At the 24 hour observation period the surviving rats has adopted a hunched stance and showed signs of lethargy.

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 oral (rat): 34 ml/kg (aqueous solution)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

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:

November 2009 - March 8, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Species: Rat, Wistar strain, Crl:WI (Han) (outbred, SPF-Quality)
- Source: Charles River Deutschland, Sulzfeld, Germany
- Number of animals: 5 males and 5 females (females were nulliparous and non-pregnant)
- Age and body weight: Young adult animals (approx. 11 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean
- Health inspection: A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from any abnormality
- Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0ºC (actual range: 18.4 – 21.7ºC), a relative humidity of 40-70% (actual range: 31 - 88%) and 12 hours artificial fluorescent light and 12 hours darkness per day
- Accommodation: Individually housed in labeled Macrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)
- Acclimatization period was at least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Macrolon cages (MIV type)
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water: Free access to tap water

IN-LIFE DATES: From: 15 December 2009 To: 29 December 2009

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure and % coverage: The test substance was applied in an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females.
- Type of wrap if used: The test substance was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D)*, successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes
- Time after start of exposure: 24 hours (using tap water)

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg (4.47 mL/kg) body weight,
expressed as surfactant content and 2612 mg/kg, expressed as solid content
Dose volume calculated as: Dose level corrected for content (g/kg) / specific gravity (g/mL).
- Concentration (if solution): N/A; the aqueous solution is tested as such

VEHICLE
Not applicable, the aqueous solution is tested as such

Duration of exposure:

24 hours

Doses:

2000 mg/kg (expressed as surfactant content);
2612 mg/kg (expressed as solid content);
5236 mg/kg (expressed as test material, the aqueous solution)

No. of animals per sex per dose:

Single dosage, on Day 1: 5 males and 5 females (females were nulliparous and non-pregnant)

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:

Mortality/Viability: Twice daily. The time of death was recorded as precisely as possible.
Body weights: Days 1 (pre-administration), 8 and 15 and at death.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
Necropsy: The animals surviving to the end of the observation period were sacrificed by an oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.

Statistics:

Not applicable

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 236 mg/kg bw

Based on:

test mat.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 612 mg/kg bw

Based on:

other: expressed as solid content

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

other: expressed as surfactant content

Mortality:

One female was found dead on Day 2. No further mortality occurred.

Clinical signs:

other: The majority of surviving animals showed one or more of the following clinical signs between Days 1 and 2: Chromodacryorrhoea, laboured respiration, rales, gasping, salivation, piloerection, hypothermia. Scales (grade 1) were seen in the treated skin-area

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the surviving animals.
Beginning autolysis was observed for the female that was found dead on Day 2.

Other findings:

Not applicable

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 dermal ( rat): > 2612 mg/kg bw (expressed as solid content)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 612 mg/kg bw

Additional information

Acute toxicity (oral)

Several studies are available, in mice and rats. They were all conducted prior to 1988, but usually followed the same general principles with some limitations in the study design and reporting. They all gave consistent results. The studies are conducted on the aqueous solution. The following table shows the results of the 3 key studies: 

Method	Reliability	Result (mL/kg) with a 20% aqueous solution	Result expressed as solid content (mg/kg bw)	Reference
Standard (equivalent to OECD 401)	2	42	42*1.17g/ml*1000*0.2= 9828	Gill, CRB ( 1977a)
Standard (equivalent to OECD 401)	2	44.5	10413	Gill, CRB ( 1977b)
Standard (equivalent to OECD 401)	2	34	7956	Gill, CRB ( 1977c)

Conclusion: >5000 mg/kg bw (in rats and mice)

Acute toxicity (dermal)

In a dermal acute toxicity study with 10 rats (limit test), a LD50 dermal of > 2612 mg/kg bw was determined.

Justification for classification or non-classification

The substance is not classified for acute toxicity in accordance with the EU Regulations as:

LD50 oral: > 5000 mg/kg

LD50 dermal: > 2612 mg/kg