1,3-di-o-tolylguanidine

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1,3-di-o-tolylguanidine

EC Number:

202-577-6

EC Name:

1,3-di-o-tolylguanidine

Cas Number:

97-39-2

Molecular formula:

C15H17N3

IUPAC Name:

1,3-di-o-tolylguanidine

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan Co. Atsugi Breeding center
- Age at study initiation: 6 weeks old
- Weight at study initiation: males = 177.4-190.2g, females = 131.4-152.7g
- Fasting period before study: yes 18 hours
- Housing: in polycarbonate cages with screen floors
- Diet (e.g. ad libitum): autoclaved sterilized solid food (CRF-1 Oriental yeast co), ad libitum
- Water (e.g. ad libitum):well water to which sodium hypochlorite had been added (2 ppm), ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):24+/-2°C
- Humidity (%):55+/-10%
- Air changes (per hr):13-15
- Photoperiod (hrs dark / hrs light): 12/12 (lighed from 7:00am-700pm)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5 % Sodium carboxymethylcellulose solution mixed with 0.1 % Tween 80

Details on oral exposure:

The administration dose was 10 mL/kg, and the administration dose for each animal was calculated based on the weight on the administration day.
The same dose of the vehicle was administered to the control group.

Doses:

0 (Vehicle), 15, 30, 60, 120 mg/kg bw

No. of animals per sex per dose:

5 rats/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations : observations on the general conditions as well as the presence of mortalities were conducted continuously for six hours after administration on the administration day (day 0 of administration), twice a day in the morning and afternoon on days 1~13 of administration, and once a day during the morning on day 14 of administration.
- Frequency of weighing: Measurements were taken before administration on the day of administration (day 0 of administration), and then on days 1, 3, 5, 7, 10 and 14 after administration. Additionally, final weight measurements were made on animals that perished.
- Necropsy of survivors performed: yes
Using ether anesthesia, exsanguination was performed on the fatalities as soon as possible after verifying the deaths, and on all of the surviving animals on after the end of the observation period. One male from the 120 mg/kg group that perished (No. 25) exhibited visual abnormalities in the lungs, and one female in each the 60 and 120 mg/kg groups (No. 70 and 72) exhibited visual abnormalities in the stomach so these were extracted and fixed in 10% neutral buffered formalin. Additionally, paraffin embedded slices were created and hematoxylin and eosin (HE) staining conducted for observation under an optical microscope.

Statistics:

From the total number of fatalities after 14 days of administration, the LD50 value was calculated for the males using the Probit method and for the females using the Van der Waerden method. Mean values and standard deviations were determined for body weights for each group. No statistically significant differences were noted.

Results and discussion

Preliminary study:

The administration amount was set based on the results of the preliminary single administration test conducted using rats [study number P020010 (not according to GLP), administration amount: 10, 30, 100 and 300 mg/kg; number of animals: 3 of each sex per group]. In this study, all of the males and females in the 100 and 300 mg/kg groups perished, and mydriasis, reduction in spontaneous activity, bradypnea and tremors were confirmed during observation of the general conditions. In this study, a maximum dose of 120 mg/kg was set where all of the cases were expected to perish, and subsequent doses were set to 60, 30 and 15 mg/kg, using a ratio of 2.

Effect levels

Effect levels 1

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

85.3 mg/kg bw

Based on:

test mat.

95% CL:

> 44.8 - < 216.9

Effect levels 2

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

56 mg/kg bw

Based on:

test mat.

95% CL:

> 40.2 - < 78

Mortality:

There are no mortality at 15 and 30 mg/kg bw and in control group.
Deaths occurred in the males and females in the 60 mg/kg (1 male and 3 females) and 120 mg/kg groups (4 males and 5 females). At 60 mg/kg, the male and 2 females died 45 minutes after administration, and one female after 30 minutes. At 120 mg/kg, 4 males and all (5) females died after 30 minutes of administration.

Clinical signs:

other: Among the survivors, one male in the 120 mg/kg group exhibited the same symptoms 15 minutes after administration as those that perished, and was observed until 6 hours after administration, and all symptoms had disappeared by the day after administration

Gross pathology:

Visual examination of the males from the fatalities in the 120 mg/kg group revealed histological mild haemorrhaging in the lung where there was slight dark red coloration. Visual examination of the females from the fatalities in the 60 and 120 mg/kg groups revealed slight dark red spots on the proventriculus, and histologically, there was mild haemorrhaging noted in the stomach layer containing the proventriculus.

Other findings:

Necropsy : Of the mortalities, 1/4 males in the 120 mg/kg group exhibited slight dark red coloration of the lung. Additionally, 1/3 females in the 60 mg/kg group and 3/5 females in the 120 mg/kg group exhibited slight dark red spots on the proventriculus. There were no changes noted in any of the survivors.

Any other information on results incl. tables

Table 1: Mortality and lD50 values in rats after a single oral administration of DOTG

 

sex	Dose (mg/kg)	Number of animals	Time after administration						Total number of dead animals	LD50 (95% confidence interval) (mg/kg)
			Minutes				hours	Day
			0	15	30	45	1-6	1-14
male	0	5	0	0	0	0	0	0	0	85.3 (44.8-216.9)
	15	5	0	0	0	0	0	0	0
	30	5	0	0	0	0	0	0	0
	60	5	0	0	0	1	0	0	1
	120	5	0	0	4	0	0	0	4
female	0	5	0	0	0	0	0	0	0	56.0 (40.2 – 78.0)
	15	5	0	0	0	0	0	0	0
	30	5	0	0	0	0	0	0	0
	60	5	0	0	1	2	0	0	3
	120	5	0	0	5				5

 

Applicant's summary and conclusion

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The LD50 values are 85.3 mg/kg bw in males and 56 mg/kg bw in females.

Executive summary:

Single oral administration of doses of 0 (control), 15, 30, 60 and 120 mg/kg of N,N’-bis (2-methyl phenyl) guanidine was performed on five male and five female Crj:CD (SD) IGS rats in each group to study the acute toxic symptoms and the lethal dose. There are no mortality at 15 and 30 mg/kg bw and in control group.

Deaths occurred in the males and females in the 60 mg/kg (1 male and 3 females) and 120 mg/kg groups (4 males and 5 females). Treatment-related clinical signs were noted as follows: mydriasis, decrease in locomotor activity, bradypnea and tremors. Among males in the 60 mg/kg group, a trend of low values for body weights was noted on day 1 after administration and throughout the observation period in females in the 60 mg/kg group.

The LD50 values are 85.3 mg/kg bw in males and 56 mg/kg bw in females.