Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1967

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:

1967

Report date:

1967

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

7 days instead of 14 days observation

Principles of method if other than guideline:

Single oral application to mice and calculation of 7 day LD50
Study performance before implementation of corresponding international guidelines, however study performance is comparable to recent guidelines to great extent.

GLP compliance:

no

Remarks:

study performance before implementation of GLP

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite

EC Number:

939-654-5

Molecular formula:

Molecular formula cannot be given as the substance is a mixture

IUPAC Name:

Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite

Test material form:

other: liquid

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Institut für industrielle und biologische Forschung, GmbH, Köln
- Age at study initiation: Not provided
- Weight at study initiation: Median weight of 23.7 g
- Fasting period before study: Approximately 17-20 hours
- Housing: Prior to the application in Plastibox cages (42x28x17 cm3) in groups of 35 animals on vermiculite. After application in Makrolon cages on wire roasting over vermiculite.
- Diet (e.g. ad libitum): Altromin R, pellet diameter 8-9mm, supplier: Altromin GmbH
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: Not provided

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 21°C
- Humidity (%): ca. 70%
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): Not provided

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.0464, 0.215, 0.464, 1.00, 1.00 and 1.00 (mL/mL) at the dose levels of 1.00, 4.64, 10.0, 21.5, 31.6 and 46.4 mL/kg bw, respectively


MAXIMUM DOSE VOLUME APPLIED: 46.4 mL/kg bw or rather 1.10 mL/mouse

Doses:

1.00 mL/kg bw; 4.64 mL/kg bw ; 10.0 mL/kg bw ; 21.5 mL/kg bw ; 31.6 mL/kg bw and 46.4 mL/kg bw

No. of animals per sex per dose:

10 for the 1.00; 4.64; and 10.0 mL/kg dose
20 for the 21.5 and 46.4 mL/kg dose
16 for the 31.6 mL/kg dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 7 days
- Frequency of observations and weighing: The mice were observed after application about 8 hours long at short intervals. Further assessments were performed several times daily until 7 days after application.
- Necropsy of survivors performed: yes
- Other examinations performed: vitality, pain reflex, cyanosis, breathing, corneal reflex, lid wide, exophtalmus, sound sensitivity, shaky walking, abdominal position, side position, running motion, clonic-tonic convulsions, ophistotonos, defecation; necropsy of deceased animals

Statistics:

The quantitative evaluation of the lethality data was performed by graphical determination of the LD50 7 days after application, in the probability grid with logarithmic shared X-axis, when using confidence limits.

Results and discussion

Effect levels

Effect levels 1

Sex:

male

Dose descriptor:

LD50

Effect level:

28.7 mL/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 as reported in study report referring to the test substance as delivered by the sponsor

Effect levels 2

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

ca. 11 000 mg/kg bw

Based on:

other: solid content

Remarks on result:

other: Recalculated LD50 referring to solid content of 39% (according to producer information) and an assumed density of roughly 1

Mortality:

Up to and including the dose level of 10.0 mL/kg bw no animal died.
In the dose group of 21.5 mL/kg bw 5% (1/20) of the animals was found death two days p.a.. In the dose groups of 31.6 and 46.4 mL/kg bw 88% (14/16) and 100% (20/20) of the animals died within 24 hours or 6 hours p.a. respectively.

Clinical signs:

other: Especially in the doses 21.5; 31.6 and 46.6 mL/kg quiet, apathetic behavior, closed eyes, diarrhea with soft faeces and decreased sound sensitivity were observed. Also in the 10.0 mL/kg dose group a number of animals showed diarrhea ( occurrence ca. 2 hou

Gross pathology:

In the stomach and small intestine of mice who died, dark reddish-brown slimy liquid was determined. The mucous membranes were dark red and translucent, and showed macroscopic signs of irritation, especially the main gastric membranes. Intubation injuries were not found.

Other findings:

- Organ weights: no data
- Histopathology: no data
- Potential target organs: no data
- Other observations: no data

Any other information on results incl. tables

Table 1. Lethality, chronological categorization (summed up, %)

Experimental Period	Dosis (mL/kg)
0 bis	1.00	4.64	10.0	21.5	31.6	46.4
20 min					6
40 min					13	35
60 min					19	60
2 h					44	70
4 h					50	90
6 h						100
24 h	0	0	0	0	88	100
2 d				5
3 d
4 d
5 d
6 d
7 d	0	0	0	5	88	100
Number of mice used	10	10	10	20	16	20

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 at 7 days was 28.7mL/kg. The LD50 for an observation period of 24 hours did practically not differ from the previous value. Recalculated LD50 referring to solid content of 39% (according to producer information) and an assumed density of roughly 1 was calculated to be ca. 11000 mg/kg bw.

Executive summary:

Acute oral toxicity (LD₅₀) of the test item was examined by gavage in male NMRI mice at doses of 1, 4.64, 10, 21.5, 31.6 and 46.4 mL/kg bw. There were no pronounced , specific toxic behavioral symptoms registered at the lower doses of 1 and 4.64 mL/kg bw. At doses ≥10 ml/kg diarrhea was seen. The died mice showed irritation of the mucous membranes of the stomach and small intestine. At the higher doses (21.5, 31.6 and 4.64 mL/kg bw/day), quiet apathetic behaviour, closed eyes and decreased sound activity were also observed.
The LD₅₀ at 7 days was 28.7 mL/kg. Recalculated LD₅₀ referring to solid content of 39% (according to producer information) and an assumed density of roughly 1 was calculated to be ca. 11000 mg/kg bw.