3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1978

Report date:

1978

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Remarks:

Study pre-dates GLP

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

EC Number:

221-326-1

EC Name:

3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

Cas Number:

3068-39-1

Molecular formula:

C27H29N2O3.Cl

IUPAC Name:

3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Albino (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Body weight prior to test material application: Males: group averages were 162 g for all groups; females: group averages were 147 g for 2 groups and 162 g for the two highest dose groups.
- Fasting period before study: Animals were deprived of food and water for 16 hours prior to test material application.
- Housing: The rats were housed in MAKROLON type III (max. 5 rats) cages consisting of a wire grid with vermiculite beneath.
- Diet: Standardised pellets for rat and mice ad libitum
- Water: ad libitum
- Acclimation period: Minimum of 3 days and maximum of 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1 °C
- Humidity: 55 ±1 0 % (relative)
- Photoperiod: 12 hour light/dark cycle. Light hours were from 07:00 to 19:00

IN-LIFE DATES: Not reported

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Bi-distilled water, 0.5 % CMC-preparation olive oil (DAB7), polyethylene glycol 400

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 10, 31.6, 100 and 316 mg/mL

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: Doses were prepared immediately before application. Emulsions and suspensions were stirred with a magnetic stirrer over the whole application period.

Doses:

100, 316, 1000 and 3160 mg/kg

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations were performed <15, 15 and 30 minutes and 1, 2, 4, 5 and 24 h after application and afterwards once daily for 14 days after application. Determination of body weight was carried out immediately before dosing as well as 7 and 14 days after application. Single weighing and the body weight related to classes of the WL24-system.
- Necropsy of survivors performed: yes

Statistics:

Probit analysis following Finney was carried out (both separately and for males and females combined).

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

449 mg/kg bw

Based on:

test mat.

95% CL:

>= 322 - <= 626

Effect levels 2

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

410 mg/kg bw

Based on:

test mat.

95% CL:

>= 265 - <= 636

Effect levels 3

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

453 mg/kg bw

Based on:

test mat.

95% CL:

>= 266 - <= 771

Mortality:

No mortalities were observed for the animals dosed at 100 mg/kg. At 316 mg/kg 2 females and 1 male died. In the 1000 mg/kg dose group 5 males and 4 females died and in the high dose group all rats died.

Clinical signs:

Clinical signs observed were apathy, diminished response to external stimulus, narrow eyelids, bristled fur, soft stools and reduced quantity of stools.

Body weight:

All surviving rats gained weight over the observation period, though at the 7 day evaluation 1 female in each of the 316 and 1000 mg/kg dose groups were considered to be highly emaciated.

Gross pathology:

The following findings were observed for rats that spontaneously died during the observation period. One rat was found with a lentil sized bleaching in the left side of the cardiac septum, 1 rat was observed with dark red focal region in the left pulmonary lobe, 1 rat had developed atelectasis across the whole lung, 2 rats had developed atelectasis in the apex of the lung and the whole gastro-intestinal tract was coloured red-violet, 3 rats had developed dark brown livers with light brown sprinklings, liver coloured yellowish, 3 rats were found to have red colourisation of all the internal tissues systemically, 1 rat was observed to be grey-red colourised and lastly 3 rats had one or double sided abdominal cryptorchism.
There were no findings from rats sacrificed at the end of the test.

Any other information on results incl. tables

Table 1: Summary of Mortalities

Dose rate (mg/kg)	Application volume (mL/kg)	Concentration (mg/mL)	n/5 Cumulative number of dead rats (timeframe)
			1 h		24 h		48 h		7 d		14 d
			M	F	M	F	M	F	M	F	M	F
100	10.0	10	0	0	0	0	0	0	0	0	0	0
316	10.0	31.6	0	0	0	1	1	1	1	1	1	2
1000	10.0	100	0	0	1	1	3	2	5	3	5	4
3160	10.0	316	0	0	3	3	3	3	5	5	5	5

Table 2: Summary of Body Weight

Dose rate (mg/kg)	Gender	Body weight of rats alive (g, geometric mean)			Mean factor for increase of body weight of surviving rats
		Pre appl.	7 d post appl.	14 d post appl.	n	0 - 7 d	n	7 - 14 d	n	0 - 14 d
100	M	162	196	237	5	1.21	5	1.21	5	1.47
	F	147	178	196	5	1.21	5	1.10	5	1.33
316	M	162	196	237	4	1.21	4	1.21	4	1.47
	F	147	162*	196	4	1.10	3	1.21	3	1.33
1000	M	162	—	—	—	—	—	—	—	—
	F	162	162*	215	2	1.00	1	1.33	1	1.33
3160	M	162	—	—	—	—	—	—	—	—
	F	162	—	—	—	—	—	—	—	—

*Rat highly emaciated

Applicant's summary and conclusion

Interpretation of results:

other: Classified as Category 4 in accordance with EU criteria

Conclusions:

Under the conditions of the study, the LD50 of the test material was found to be 449 mg/kg bw for males and females combined (95 % confidence limit 322 to 626 mg/kg bw). The LD50 for males was 410 mg/kg bw (95 % CL 265 to 636 mg/kg bw) and for females was 453 mg/kg bw (95 % CL 266 to 771 mg/kg bw).

Executive summary:

A study was conducted to assess the acute oral toxicity potential of the test material in the rat using methodology equivalent to the standardised guideline OECD 401.

Groups of 5 female and 5 male rats were exposed the test material by gavage at dose levels of 100, 316, 1000 and 3160 mg/kg using a vehicle of bi-distilled water, 0.5 % CMC-preparation olive oil (DAB7) and polyethylene glycol 400. The animals were observed for a 14 day period for mortality and clinical signs. Necropsies were performed for animals that died spontaneously and for animals that were euthanised at the end of the study.

No mortalities were observed for the animals dosed at 100 mg/kg. At 316 mg/kg 2 females and 1 male died. In the 1000 mg/kg dose group 5 males and 4 females died and in the high dose group all rats died. Clinical signs observed were apathy, diminished response to external stimulus, narrow eyelids, bristled fur, soft stools and reduced quantity of stools. All surviving rats gained weight over the observation period, though at the 7 day evaluation 1 female in each of the 316 and 1000 mg/kg dose groups were considered to be highly emaciated.

The following findings were observed for rats that spontaneously died during the observation period. One rat was found with a lentil sized bleaching in the left side of the cardiac septum, 1 rat was observed with dark red focal region in the left pulmonary lobe, 1 rat had developed atelectasis across the whole lung, 2 rats had developed atelectasis in the apex of the lung and the whole gastro-intestinal tract was coloured red-violet, 3 rats had developed dark brown livers with light brown sprinklings, liver coloured yellowish, 3 rats were found to have red colourisation of all the internal tissues systemically, 1 rat was observed to be grey-red colourised and lastly 3 rats had one or double sided abdominal cryptorchism. There were no findings from rats sacrificed at the end of the test.

Under the conditions of the study, the LD50 of the test material was found to be 449 mg/kg bw for males and females combined (95 % confidence limit 322 to 626 mg/kg bw). The LD50 for males was 410 mg/kg bw (95 % CL 265 to 636 mg/kg bw) and for females was 453 mg/kg bw (95 % CL 266 to 771 mg/kg bw).