1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

9 July to 5 September 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

EC Number:

855-905-0

Cas Number:

1326716-46-4

Molecular formula:

C24H40N2O6S2 (C10H24N2 • 2 C7H8O3S)

IUPAC Name:

1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

Test material form:

solid

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS B.V., Inc, Postbus 6174, 5960 AD Horst / The Netherlands
- Age (beginning of treatment): 6 - 13 weeks
- Body weight at day 0 of the study: 153.8 - 252.6 g
- Identification: The animals were distributed into the test groups at random. If possible, all animals belonging to the same experimanetal group were kept in one cage. The animals were individually marked. A colour-coded card was prepared for each project, giving details of the test type, project number, treatment start, dose level, sex and number of animals.
- Acclimatization: At least 5 days prior to the start of dosing under test conditions after health examination. Only animals without any visible signs off illness were used for the study.
- Fasting period before study: overnight
- Housing: groups of one to five rats (of the same sex and dose group)
- Cage Type: Makrolon Type IV, with wire mesh top
- Feed: 2018C Teklad Global 18% protein rodent diet (certified), ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approximately 20-27 °C
- Humidity (%): approximately 45-72%
- Photoperiod (hrs dark / hrs light): artificial light 6.00 a.m. - 6.00 p.m. (12 hrs dark / 12 hrs light)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: various
- Amount of vehicle (if gavage): various
- Justification for choice of vehicle: commonly used solvent

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: Based on available information on the toxicity of the test item, 300 mg/kg was chosen at initial starting dose, since a severe toxicity which may necessitate human euthanasia was not expected at this dose level.

Doses:

300, 2000 and 5000 mg/mL

No. of animals per sex per dose:

SIGHTING STUDY:
300 mg/kg bw: 1 female
2000 mg/kg bw: 1 female
5000 mg/kg bw: 1 female

MAIN STUDY:
2000 mg/kg bw: 4 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical observations and inspections for morbidity/mortality were performed at least three times within the first six hours after application (i.e. 30 minutes and 1 hour, 2 hours and 4 hours after dosing, thereafter at least once daily for 14 days. All surviving animals were observed for 14 days after dosing. Body weights were recorded on day 0 (prior to dosing), day 7, and 14, or (if applicable) at death (unscheduled).
- Necropsy of survivors performed: yes
- Clinical signs including body weight: Clinical observations and inspections for morbidity / mortality were performed at least three times within the first six hours after application (i.e., 30 minutes and 1 hour, 2 hours and 4 hours after dosing), thereafter at least once daily for 14 days. All surviving animals were observed for 14 days after dosing.
The nature and severity, where appropriate, of the clinical signs and the time were recorded at each observation for all individual animals. If applicable, the time of death/humane kill was recorded as precisely as possible. Observations included changes in the skin and fur, eyes and mucous membranes, and respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behavior pattern. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 2 000 - <= 5 000 mg/kg bw

Based on:

other: Clevios K Primer W8 dry

Remarks on result:

other: CL not determined

Mortality:

SIGHTING STUDY:
For each dose level one animal was treated:
at dose level 300 mg/kg bw: the animal treated with this dose level in the sighting study survived until the end of ebservation period (day 14)
at dose level 2000 mg/kg bw: the animal treated with this dose level in the sighting study survived until the end of ebservation period (day 14)
at dose level 5000 mg/kg bw: the female treated with this dose level was found dead approximately 4 hours after application

MAIN STUDY
Based on the results obtained in the sighting study, a further group of four animals was treated with a dose level of 2000 mg/kg bw.
One female had to be humanely sacrificed on day 1 after the application. Clinical signs prior to sacrifice comprised hunched posture, piloerection, sunken flanks, decreased activity, tiptoe gait, diarrhoea, abdominal tension, darkened eyes, and ataxia.

Clinical signs:

other: SIGHTING STUDY (one animal per dose level): Clinical signs at dose level 300 mg/kg bw: no clinical signs of reaction to treatment throughout this study 2000 mg/kg bw: apart from transient piloerection (2 h after application) the animal did not show any cl

Gross pathology:

Animals found dead:
The animal dosed at 5000 mg/kg bw was found dead approximately 4 hours after application.
Upon necropsy, the animal showed a wet nose, an acute liver congestion, furthermore the stomach was bloated, dark red stained and filled with liquid, the small intestine was filled with liquid, the caeucum was atrophied, and the lungs were filled with white foam (indicative of an acute toxic lung edema).
One animal dosed at 2000 mg/kg bw had to be humanly sacrified on day 1 after the application. Clinicla signs proir to sacrifice comprised hunched posture, piloerection, sunken flanks, decreased activity, tiptoe gait, diarrhoea, abdominal tension, darkened eyes, and ataxia.

Surviving animals (necropsy at day 14):
No macroscopic abnormalities in the animal dosed at 300 mg/kg bw or in surviving animals dosed at 2000 mg/kg bw and euthanatized on day 14.

Any other information on results incl. tables

Individual Body Weights and Body Weight changes

Dose Level mg/kg bw	Animal Number and Sex	Body Weight (g) at Day			Body Weight Gain (g) During Week		Body Weight Gain (%) During Week
		0	7	14	1	2	1	2
300	Female 1	171.7	193.6	201.3	21.9	7.7	12.8	4.0
2000	Female 1	153.8	184.0	207.9	30.2	23.9	19.6	13.0
	Female 2	245.0	269.9	277.0	24.9	7.1	10.2	2.6
	Female 3	219.0	242.0	244.4	23.0	2.4	10.5	1.0
	Female 4	252.6	animal was sacrified on day 1 after application
	Female 5	233.2	248.7	255.9	15.5	7.2	6.6	2.9
5000	Female 1	237.2	animal was found dead approx. 4 h after application

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

In an acute oral toxicity test on female rats, conducted to GLP and according to OECD Test Guideline 420, the acute median lethal oral dose (LD50) to rats of Clevios K Primer W8 dry was demonstrated to be between 2000 and 5000 mg/kg body weight.

Executive summary:

The acute oral toxicity of 1,10 -Decanediamine, 4 -methylbenzenesulfonate (1:2) (Trade name: Clevios K Primer W8 dry) was assessed using the fixed dose procedure. The study was performed to GLP and according to OECD Test Guideline 420.

Female WISTAR rats were treated with a single oral (gavage) dose of test material (in corn oil)

- in a Sighting Study at 300 mg/kg bw (1 animal), at 2000 mg/kg bw (1 animal) and at 5000 mg/kg bw (1 animal) and

- in a Main Study at 2000 mg/kg bw (4 animals)

before a 14 -day observation period.

Anmials were checked for clinical signs of toxicity and effects on body weight. Rats were subject to cross necropsy upon death or terminal killing.

The animal treated with 5000 mg/kg bw was found dead approximately 4 hours after application. Of the in total five animals treated with 2000 mg/kg bw, one animal was sacrified on day 1 after application.

Under the conditions of this study, the acute oral LD50 was demonstrated to be between 2000 and 5000 mg/kg bw. 1,10 -Decanediamine, 4 -methylbenzenesulfonate (1:2) is therefore classified as Category 5 for acute toxicity via the oral route under the EU CLP regulation.