reaction products of 1 mole of Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs and 1 mole of cyclohexylamine

Endpoint summary

Description of key information

An acute oral toxicity study in rats (OECD 423) is available for Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine. The acute oral LD50 value of this substance was found to be greater than 2000 mg active ingredient/kg bw in female Crl:WIrats.

There are no data for dermal route and inhalation.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 March 2017 to 13 April 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

OECD Guidelines for Testing of Chemicals No. 423. Acute Oral Toxicity – Acute Toxic Class Method. Adopted: 17 December 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

EPA Health Effects Test Guidelines (OPPTS 870.1100), United States, EPA 712-C-02-190 (2002)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species and strain: Crl:WI Wistar rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals, 3 animals / group
Sex: Female, nulliparous and non-pregnant
Age of animals at dosing: Young healthy adult rats, 9 weeks old
Body weight at treatment: 196 – 216 g
Acclimation period: at least 13 days

Husbandry
Animal health: Only healthy animals were used for the test. The health status was certified by the staff Veterinarian.
Number of animal room: 242/3
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: LIGNOCEL 3/4-S certified wooden chips (produced by J. Rettenmaier & Söhne GmbH + CO. KG 73494, Holzmühle, Rosenberg, Germany) was available to animals during the study.
Nesting: ARBOCEL crinklets natural nest building material (produced by J. Rettenmaier & Söhne GmbH + CO. KG 73494, Holzmühle, Rosenberg, Germany) was available to animals during the study.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 19.6 – 25.0°C
Relative humidity: 30 – 58%
Ventilation: 15 – 20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

Food and Water Supply
Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany (Batch no.: 484 14771, expiry date: 30 June 2017 and Batch no.: 285 17890, expiry date: 31 August 2017), ad libitum, except for the night before treatment. Tap water from the municipal supply, as for human consumption was available from a 500 ml bottle, ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly.

Animal Identification
Animals were individually identified using numbers written on the tail with an indelible pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.'s Master File, for each animal allocated to the treatment groups. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Formulation
The test item was administered undiluted for dose level of 2000 mg active ingredient/kg bw at a constant concentration adjusting for the specific gravity of the test material. This method was chosen considering the highest permissible dose volume (20 ml/kg bw) by the guideline.

Justification of the dose:
The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. Based on the preliminary toxicological information by the Sponsor, limit dose of 2000 mg active ingredient/kg bw was selected as a starting dose.
Initially, 3 female animals were treated with Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine at dose level of 2000 mg active ingredient/kg bw. Only 1 out of 3 animals died in this group, therefore further 3 animals were treated at the dose level of 2000 mg active ingredient/kg bw. Since only one animal died in this second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).

Doses:

2000 mg active ingredient/kg bw.

No. of animals per sex per dose:

3 animals per group (6 in total)

Control animals:

no

Details on study design:

Procedure
A single oral gavage administration was followed by a 14-day observation period. On the night before treatment, the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment.
Single oral gavage was administered using a constant concentration at the tested dose level of 2000 mg active ingredient/kg bw. Dose volume was 17.7 ml/kg bw considering the 1.129 g/mL density and the correction factor 10.

OBSERVATIONS
Clinical Observations
Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter or until death. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement
The body weight was recorded on Days -1, 0 (before treatment), 7 and 14 (before necropsy) or at death.

NECROPSY
Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Release®; Lot No.: 106075, Expiry Date: 31 July 2018, Produced by: Wirtschaftsgenossenschaft deutscher Tierärzte eG; Siemensstr. 14, 30827 Garbsen, Germany). After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross pathological changes were recorded for each animal on the post mortem record sheets and the animals were discarded.

Statistics:

The method used was not intended to allow the calculation of a precise LD50 value.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

Two out of 6 animals were found dead during the study at dose level of 2000 mg active ingredient/kg bw (1 animal in Group 1 and another in Group 2, both on Day 1).

Clinical signs:

At the dose level of 2000 mg active ingredient/kg bw, the following symptoms were observed: hunched back (6/6 animals, Day 0-2), straub like tail position (6/6 animals, Day 0), piloerection (4/6 animals, Day 0-1), irritability (3/6 animals, Day 0-1), slight activity decrease (3/6 animals, Day 0-1) and reddish coloured discharge from nose (1/6 animals, Day 1).
From Day 3 all surviving animals were symptom-free.

Body weight:

There were no effects on body weights or body weight gains that could be attributed to treatment with Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine.

Gross pathology:

Yellowish liquid observed in the digestive contents of the stomach and/or duodenum/jejunum was considered to be treatment-related.
Dry red material observed in the fur of periorbital area (bilateral) and diffuse dark/red discolorations all lobes of the collapsed lungs found at necropsy, were considered to be non-treatment-related. Discoloured lungs are frequently seen in found dead rats.
There was no evidence of the macroscopic observations in animals subjected to the necropsy on Day 14.

CLINICAL OBSERVATIONS

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0                                                                                        SEX: FEMALE

Cage No.	Animal Number	Observations	Observation days													Frequency
			0						1	2	3	4	5	6	7-14
			30’	1h	2h	3h	4h	6h
1	7335#	Symptom Free	+	+	+	+	-	-								4/6
		Irritability	-	-	-	-	+	+								2/6
		Hunched back	-	-	-	-	+	+								2/6
		Piloerection	-	-	-	-	-	+								1/6
		Straub like tail position	-	-	-	-	+	+								2/6
		Found dead	-	-	-	-	-	-	+							-
	7336	Symptom Free	+	+	+	+	-	-	-	-	+	+	+	+	+	16/20
		Activity decreased	-	-	-	-	-	-	1	-	-	-	-	-	-	1/20
		Hunched back	-	-	-	-	+	+	+	+	-	-	-	-	-	4/20
		Piloerection	-	-	-	-	-	-	+	-	-	-	-	-	-	1/20
		Discharge, coloured (reddish, nose)	-	-	-	-	-	-	+	-	-	-	-	-	-	1/20
		Straub like tail position	-	-	-	-	+	+	-	-	-	-	-	-	-	2/20
	7337	Symptom Free	+	+	+	+	-	-	-	+	+	+	+	+	+	17/20
		Activity decreased	-	-	-	-	-	-	1	-	-	-	-	-	-	1/20
		Hunched back	-	-	-	-	+	+	+	-	-	-	-	-	-	3/20
		Piloerection	-	-	-	-	-	-	+	-	-	-	-	-	-	1/20
		Straub like tail position	-	-	-	-	+	+	-	-	-	-	-	-	-	2/20
2	7338	Symptom Free	+	+	-	-	-	-	-	+	+	+	+	+	+	15/20
		Irritability	-	-	+	+	+	+	+	-	-	-	-	-	-	5/20
		Hunched back	-	-	+	+	+	+	+	-	-	-	-	-	-	5/20
		Straub like tail position	-	-	-	+	+	+	-	-	-	-	-	-	-	3/20
	7303#	Symptom Free	+	-	-	-	-	-								1/6
		Activity decreased	-	-	-	-	-	1								1/6
		Hunched back	-	-	+	+	+	+								4/6
		Piloerection	-	+	+	+	+	+								5/6
		Straub like tail position	-	-	-	+	+	+								3/6
		Found dead	-	-	-	-	-	-	+							-
	7304	Symptom Free	+	-	-	-	-	-	-	+	+	+	+	+	+	14/20
		Irritability	-	-	+	+	+	+	-	-	-	-	-	-	-	4/20
		Hunched back	-	+	+	+	+	+	+	-	-	-	-	-	-	6/20
		Straub like tail position	-	-	-	+	+	+	-	-	-	-	-	-	-	3/20

Remarks:              + = present                                                           - = absent

                               h = hour                                                ‘ = minute

                               # = Found dead

Frequency of observations = number of occurrences of observation / total number of observations

Severities = 1 = slight/Small/Few; 2 = Moderate/Medium; 3 = Marked/Large/Many

 

BODY WEIGHT DATA

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0                                                    SEX: FEMALE

Cage No.	Animal Number	Body Weight (g)				Day/Body Weight (g) Death	Body Weight Gain (g)
		Days
		-1	0	7	14		-1-0	0-7	7-14	-1-14
1	7335#	226	211	-	-	1/205	-15	-	-	-
	7336	225	214	251	270	-	-11	37	19	45
	7337	225	211	224	242	-	-14	13	18	17
2	7338	206	196	215	223	-	-10	19	8	17
	7303#	215	206	-	-	1/204	-9	-	-	-
	7304	227	216	236	247	-	-11	20	11	20
Mean:		220.7	209.0	231.5	245.5	-	-11.7	22.3	14.0	24.8
Standard deviation:		8.4	7.2	15.6	19.3	-	2.3	10.3	5.4	13.6

- = No data

# = Found dead

 

NECROPSY FINDINGS

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0                             SEX: FEMALE

Cage No.	Animal Number	Necropsy Date/ Necropsy Day	External Observations	Internal Observations	Organ/Tissue
1	7335#	29 March 2017 Day 1	Fur: Material, dry, red, periorbital area, bilateral	Collapsed	Lungs
				Discoloration, dark red, diffuse, all lobes
				Digestive Content: Material, liquid, yellowish	Stomach and Duodenum and Jejunum
	7336	11 April 2017 Day 14	No external observations recorded	No internal observation recorded	Not applicable
	7337	11 April 2017 Day 14	No external observations recorded	No internal observation recorded	Not applicable
2	7338	13 April 2017 Day 14	No external observations recorded	No internal observation recorded	Not applicable
	7303#	31 March 2017 Day 1	No external observations recorded	Collapsed	Lungs
				Discolouration, dark red. Diffuse, all lobes
				Digestive Content: Material, liquid, yellowish	Stomach
	7304	13 April 2017 Day 14	No external observations recorded	No internal observations recorded	Not applicable

# = Found dead

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine was found to be above 2000 mg active ingredient/kg bw in female Crl:WI rats.
According the GHS criteria, Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine can be ranked as "Category 5" for acute oral exposure.

Executive summary:

The single-dose oral toxicity of Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris) in Crl:WI Wistar rats.

 

Two groups of 3 female Crl:WI rats were treated with the test item at a dose level of 2000 mg active ingredient/kg body weight (bw) (Group 1 and Group 2).

 

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was administered undiluted for dose level of 2000 mg active ingredient/kg bw at a constant concentration adjusting for the specific gravity of the test material.

 

Initially, three females (Group 1) were treated at a dose level of 2000 mg active ingredient/kg bw. Only 1 out of 3 animals died in this group, therefore further 3 animals (Group 2) were treated at the dose level of 2000 mg active ingredient/kg bw.

Since only one animal died in this second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).

 

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter or until death. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy) or at death. All animals were subjected to a necropsy and a macroscopic examination.

 

The results of the study were summarised as follows:

Mortality

Two out of 6 animals were found dead during the study at dose level of 2000 mg active ingredient/kg bw (1 animal in Group 1 and another in Group 2, both on Day 1).

 

Clinical Observations

At the dose level of 2000 mg active ingredient/kg bw, the following symptoms were observed: hunched back (6/6 animals, Day 0-2), straub like tail position (6/6 animals, Day 0), piloerection (4/6 animals, Day 0-1), irritability (3/6 animals, Day 0-1), slight activity decrease (3/6 animals, Day 0-1) and reddish coloured discharge from nose (1/6 animals, Day 1).

From Day 3 all surviving animals were symptom-free.

 

Body Weight and Body Weight Gain

There were no effects on body weights or body weight gains that could be attributed to treatment with Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. With cyclohexylamine.

 

Necropsy

Yellowish liquid observed in the digestive contents of the stomach and/or duodenum/jejunum was considered to be treatment-related.

Dry red material observed in the fur of periorbital area (bilateral) and diffuse dark/red discolorations all lobes of the collapsed lungs found at necropsy, were considered to be non-treatment-related. Discoloured lungs are frequently seen in found dead rats.

There was no evidence of the macroscopic observations in animals subjected to the necropsy on Day 14.

 

Conclusion:

Under the conditions of this study, the acute oral LD50 value of the test item Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine was found to be greater than 2000 mg active ingredient/kg bw in female Crl:WI rats.

 

According to the GHS criteria, Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine can be ranked as "Category 5" for acute oral exposure.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Good. The study statisfies to the OECD guideline and BPLs requirements.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The single-dose oral toxicity of Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris) in Crl:WI Wistar rats.

Two groups of 3 female Crl:WI rats were treated with the test item at a dose level of 2000 mg active ingredient/kg body weight (bw). A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again three hours after the treatment. The test item was administered undiluted for dose level of 2000 mg active ingredient/kg bw at a constant concentration adjusting for the specific gravity of the test material. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter or until death. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy) or at death. All animals were subjected to a necropsy and a macroscopic examination.

Two out of 6 animals were found dead during the study at dose level of 2000 mg active ingredient/kg bw (1 animal in Group 1 and another in Group 2, both on Day 1). Hunched back (6/6 animals, Day 0-2), straub like tail position (6/6 animals, Day 0), piloerection (4/6 animals, Day 0-1), irritability (3/6 animals, Day 0-1), slight activity decrease (3/6 animals, Day 0-1) and reddish coloured discharge from nose (1/6 animals, Day 1) were noted. From Day 3, all surviving animals were symptom-free.

There were no effects on body weights or body weight gains that could be attributed to treatment with Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine.

At necropsy of the animals found dead, yellowish liquid was observed in the digestive contents of the stomach and/or duodenum/jejunum and considered to be treatment-related.

Dry red material observed in the fur of periorbital area (bilateral) and diffuse dark/red discolorations in all lobes of the collapsed lungs were considered to be non-treatment-related since discoloured lungs are frequently seen in found dead rats.

There were no recordings of macroscopic observations in animals subjected to the necropsy on Day 14.

 

In conclusion, under the conditions of this study, the acute oral LD50value of the test item Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with cyclohexylamine was found to be greater than 2000 mg active ingredient/kg bw in female Crl:WI rats.

Justification for classification or non-classification

According to the criteria laid down in EU regulation (EC) n° 1272/2008/EC (CLP) and EU directive67/548/EEC, Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.,compds. with cyclohexylamine is not classified for acute toxicity.