Methyl toluene-4-sulphonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 May 2017 to 06 June 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

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:

yes

Remarks:

See "Any other information" for details

Test guideline 2

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:

yes

Remarks:

See "Any other information" for details

Test guideline 3

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-98-190 (1998)

Deviations:

yes

Remarks:

See "Any other information" for details

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Methyl toluene-4-sulphonate

EC Number:

201-283-5

EC Name:

Methyl toluene-4-sulphonate

Cas Number:

80-48-8

Molecular formula:

C8H10O3S

IUPAC Name:

methyl 4-methylbenzene-1-sulfonate

Test material form:

solid

Details on test material:

Name: PTSM
Chemical name: Methyl toluene-4-sulphonate
CAS number: 80-48-8
Batch/Lot Number: 609271
Description: White to pale yellow solid
Expiry Date: 01 March 2018
Purity: 99.7%
Storage Conditions: Room temperature
Safety precautions: Routine safety precautions (lab coat, gloves, safety glasses, face mask) for unknown materials were applied to assure personnel health and safety.

Specific details on test material used for the study:

No further details specified in the study report.

Test animals

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: 9 animals, 3 animals/group
Sex: Female, nulliparous and non-pregnant
Age of animals at dosing: Young healthy adult rats, 8-10 weeks old
Body weight at treatment: 194 – 234 g
Acclimatisation period: At least 5 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: 245/8
Housing: 3 animals / cage
Cage type: Type II. polypropylene/polycarbonate
Bedding and nesting: “Lignocel 3/4-S Hygienic Animal Bedding” and “Arbocel crinklets natural” nesting material produced by J. Rettenmaier & Söhne GmbH + Co.KG (D-73494 Rosenberg, Germany) were available to animals during the study.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 19.1 – 25.9 °C
Relative humidity: 31 – 65 %
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.
The temperature and relative humidity were recorded twice daily during the acclimatisation period and throughout the study.

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 number: 285 17890, expiry date: 31 August 2017), ad libitum, and tap water from the municipal supply, as for human consumption 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, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A. u. 36., Hungary).

Animal Identification
Animals were individually identified using numbers written on the tail with an indelible marker 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.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

The test item was freshly formulated at a concentration of 30 or 200 mg/mL in the vehicle, in the Pharmacy of CiToxLAB Hungary Ltd. on the day of administration.
The formulation container was stirred continuously up to finishing the treatment.

Vehicle: PEG 400 (polyethylene glycol 400)
Manufacturer: Sigma-Aldrich
Batch number: BCBS1795V
Expiry date: 31 May 2018

Doses:

Justification of the dose:
A limit test was not performed as the acute oral LD50 value of the test substance was expected to be 341 mg/kg bw in rats, based on the information provided by the Sponsor. A starting dose of 300 mg/kg bw was selected by the Study Director in agreement with the Sponsor.
Initially three animals were treated at the starting dose. As only one animal died, the dose of 300 mg/kg bw was repeated with three additional animals. As no animal died in the additional group at 300 mg/kg bw, the third group received a higher dose (2000 mg/kg bw). As each animal died in this 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).

No. of animals per sex per dose:

3 animals/group

Control animals:

no

Details on study design:

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.

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, where possible. 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 the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter, where possible.

NECROPSY
Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Release, 30% pentobarbital sodium; Lot number: 106075, Expiry date: 31 July 2018, Produced by: Wirtschaftsgenossenschaft deutscher Tierärzte eG, Germany). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

Statistics:

The method used was not intended to allow the calculation of a precise LD50 value.
The test item was ranked into categories of Globally Harmonized Classification System (GHS (rev. 6) 2015). Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

PTSM caused mortality in 1 of 6 animals at a dose level of 300 mg/kg bw and in 3 of 3 animals at a dose level of 2000 mg/kg bw.

Clinical signs:

other: In the groups treated at a dose level of 300 mg/kg bw, hunched back (6/6), slight to moderate decreased activity (4/6), piloerection (3/6), slight to moderate incoordination (2/6) and wasted condition (2/6) were recorded. In the group treated at a dose le

Gross pathology:

Red or dark red diffuse discoloration of all lobes of the lung was found in the rats that died during the observation period.
No macroscopic findings were noted in the surviving animals at necropsy dosed at 300 mg/kg bw and terminated on Day 14.

Other findings:

No further findings specified in the study report.

Any other information on results incl. tables

CLINICAL OBSERVATIONS

DOSE LEVEL: 300 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	8789	Symptom free	+	+	-	-	-	-	-	-	-	-	-	+	+	11/20
		Activity decreased	-	-	1	1	1	1	1	2	2	2	1	-	-	9/20
		Hunched back	-	-	+	+	+	+	+	+	+	+	+	-	-	9/20
		Incoordination	-	-	-	-	-	-	-	1	-	-	-	-	-	1/20
		Piloerection	-	-	+	+	+	+	+	+	+	+	-	-	-	8/20
		Wasted	-	-	-	-	--	-	-	-	+	+	-	-	-	2/20
	8790	Symptom free	+	+	-	-	-	-	-	+	+	+	+	+	+	15/20
		Activity decreased	-	-	1	1	1	1	-	-	-	-	-	-	-	4/20
		Hunched back	-	-	+	+	+	+	+	-	-	-	-	-	-	5/20
		Piloerection	-	-	+	+	+	+	+	-	-	-	-	-	-	5/20
	8791#	Symptom free	+	+	-	-	-	-	-	-	-	-				2/10
		Activity decreased	-	-	1	1	1	1	1	2	2	2				8/10
		Hunched back	-	-	+	+	+	+	+	+	+	+				8/10
		Incoordination	-	-	-	-	-	-	-	2	-	-				1/10
		Piloerection	-	-	+	+	+	+	+	+	+	+				8/10
		Wasted	-	-	-	-	-	-	-	-	+	+				2/10
		Found dead	-	-	-	-	-	-	-	-	-	-	+			-
2	8792	Symptom free	+	+	+	-	-	-	+	+	+	+	+	+	+	17/20
		Hunched back	-	-	-	+	+	+	-	-	-	-	-	-	-	3/20
	8793	Symptom free	+	+	+	-	-	-	+	+	+	+	+	+	+	17/20
		Hunched back	-	-	-	+	+	+	-	-	-	-	-	-	-	3/20
	8794	Symptom free	+	+	+	-	-	-	+	+	+	+	+	+	+	17/20
		Activity decreased	-	-	-	-	1	1	-	-	-	-	-	-	-	2/20
		Hunched back	-	-	-	+	+	+	-	-	-	-	-	-	-	3/20
DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0															SEX: FEMALE
5	9311#	Activity decreased	1	2												2/2
		Hunched back	+	+												2/2
		Prone position	-	+												1/2
		Incoordination	1	-												1/2
		Piloerection	-	+												1/2
		Found dead	-	-	+											-
	9312#	Activity decreased	1	1	2	2										4/4
		Hunched back	+	+	+	+										4/4
		Prone position	-	-	-	+										1/4
		Incoordination	1	1	1	-										3/4
		Piloerection	-	+	+	+										3/4
		Found dead	-	-	-	-	+									-
	9313#	Activity decreased	1	2												2/2
		Hunched back	+	+												2/2
		Incoordination	1	1												2/2
		Piloerection	-	+												1/2
		Found dead	-	-	+											-

Remarks:              + = present           - = absent

                          h = hour              ‘ = minute

                          # = Found dead

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

                           Severities: 1 = slight/small/few, 2 = moderate/medium, 3 = marked/large/many

BODY WEIGHT DATA

DOSE LEVEL: 300 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	8789	212	194	191	237	-	-18	-3	46	25
	8790	220	201	244	255	-	-19	43	11	35
	8791	215	194	-	-	5/140	-21	-	-	-
2	8792	217	198	235	248	-	-19	37	13	31
	8793	233	218	238	259	-	-15	20	21	26
	8794	222	202	248	257	-	-20	46	9	35
Mean:		219.8	201.2	231.2	251.2	-	-18.7	28.6	20.0	30.4
Standard deviation:		7.4	8.9	23.0	9.0	-	2.1	20.3	15.2	4.8
DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0									SEX: FEMALE
5	9311#	242	234	-	-	-	-8	-	-	-
	9312#	223	213	-	-	-	-10	-	-	-
	9313#	228	213	-	-	-	-15	-	-	-
Mean:		231.0	220.0	-	-	-	-11.0	-	-	-
Standard deviation:		9.8	12.1	-	-	-	3.6	-	-	-

- = No data

# = Found dead

 

NECROPSY FINDINGS

DOSE LEVEL: 300 mg/kg bw, Treatment on Day 0					SEX: FEMALE
Cage No.	Animal Number	Necropsy Date/ Necropsy Day	External Observations	Internal Observations	Organ/Tissue
1	8789	23 May 2017 Day 14	No external observations recorded	No internal observations recorded	Not applicable
	8790	23 May 2017 Day 14	No external observations recorded	No internal observations recorded	Not applicable
	8791#	14 May 2017 Day 5	No external observations recorded	Non collapsed	Lungs
				Discoloration, red, diffuse, all lobes
2	8792	30 May 2017 Day 14	No external observations recorded	No internal observations recorded	Not applicable
	8793	30 May 2017 Day 14	No external observations recorded	No internal observations recorded	Not applicable
	8794	30 May 2017 Day 14	No external observations recorded	No internal observations recorded	Not applicable
DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0					SEX: FEMALE
5	9311#	06 June 2017 Day 0	No external observations recorded	Collapsed	Lungs
				Discoloration, red, diffuse, all lobes
	9312#	06 June 2017 Day 0	No external observations recorded	Non collapsed	Lungs
				Dark discoloration, red, diffuse, all lobes
	9313#	06 June 2017 Day 0	No external observations recorded	Non collapsed	Lungs
				Discoloration, red, diffuse, all lobes

# = Found dead

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item PTSM was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.
According the GHS criteria, PTSM can be ranked as "Category 4" for acute oral exposure.

Executive summary:

The single-dose oral toxicity of PTSM 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 rats.

 

Two groups of three female Crl:WI rats were treated with the test item at a dose level of 300 mg/kg bw (Group 1 and Group 2) and one group of three female Crl:WI rats at a dose level of 2000 mg/kg bw (Group 3).

 

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 formulated in PEG 400 at a concentration of 30 or 200 mg/mL at a dose volume of 10 mL/kg bw.

 

Initially, three females (Group 1) were treated at a dose level of 300 mg/kg bw. As only one animal was found dead, a confirmatory group (Group 2) was treated at the same dose level. As no animal died in the confirmatory group, the next dose level was 2000 mg/kg bw. As all animals were found dead in the group treated at a dose level of 2000 mg/kg bw (Group 3), no further testing was required according to OECD 423 and 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, where possible. Body weight was measured on Days -1, 0 and 7 and 14 (before necropsy), where possible. All animals were subjected to a necropsy and a macroscopic examination.

 

RESULTS

Mortality

PTSM caused mortality in 1 of 6 animals at a dose level of 300 mg/kg bw and in 3 of 3 animals at a dose level of 2000 mg/kg bw.

 

Clinical Observations

In the groups treated at a dose level of 300 mg/kg bw, hunched back (6/6), slight to moderate decreased activity (4/6), piloerection (3/6), slight to moderate incoordination (2/6) and wasted condition (2/6) were recorded.

In the group treated at a dose level of 2000 mg/kg bw, slight to moderate decreased activity (3/3), hunched back (3/3), slight incoordination (3/3), piloerection (3/3) and prone position (2/3) were seen before death.

 

Body Weight and Body Weight Gain

Treatment related body weight loss was observed in one of the surviving animals following treatment, but the animal recovered the lost weight during the second week of the observation period. The body weight changes of the animals died during the study could not be evaluated. The body weight of the other treated animals during the study showed no indication of a test item-related effect.

 

Macroscopic Findings

Red or dark red diffuse discoloration of all lobes of the lungs was found in the rats that died during the observation period.

No macroscopic findings were noted in the surviving animals at necropsy dosed at 300 mg/kg bw and terminated on Day 14.

 

CONCLUSION

Under the conditions of this study, the acute oral LD50 value of the test item PTSM was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

 

According the GHS criteria, PTSM can be ranked as "Category 4" for acute oral exposure.