Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., ammonium salts

Endpoint summary

Description of key information

Acute Toxicity: Oral: LD50 = >300 - <2,000 mg/kg bw; OECD 423; Anon., 2017

Acute Toxicity: Inahaltion: Waiver

Acute Toxicity: Dermal: LD50 = > 2,000 mg/kg bw; OECD 402; A Poole., 2018

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:

25 April 2017 - 18 January 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted in accordance with international guidelines and in accordance with GLP. All guideline validity criteria were met.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity:
- Specific activity:
- Locations of the label:
- Expiration date of radiochemical substance:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
- Stability under test conditions:
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

OTHER SPECIFICS:

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Margate, UK
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 8-9 weeks
- Weight at study initiation: All within ±20 % of the mean value
- Fasting period before study: Yes, on the evening prior to dosing.
- Housing: The animals were housed in groups of up to five during the acclimatisation period in cages that conform to the 'Code of Practice for the Housing and Care of Animals Bred, Supplied or Used for Scientific Purposes’ (Home Office, London, 2014). From the day prior to dosing (Day-1), the rats were housed in groups of three in similar cages. Bedding was provided on a weekly basis to each cage by use of clean European soft wood bedding (Datesand Ltd., Manchester, UK). Each batch of bedding was analysed for specific constituents and contaminants. No contaminants were present in bedding at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Diet (e.g. ad libitum): Throughout the study the animals had access to 5LF2 EU Rodent Diet 14%, which was freely available to the animals at all times, except for a period of fasting from the evening of the day prior to dosing (Day-1) until approximately 3 hours after dosing. Each batch of diet had been analysed for specific constituents and contaminants by the manufacturer. No contaminants were present in diet at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Water (e.g. ad libitum): Mains water was provided ad libitum via cage mounted water bottles. The water was periodically analysed for specific contaminants. No contaminants were present in water at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Acclimation period: 7 to 10 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 ºC
- Humidity (%): 45-65 %
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: From: 25 April 2017 To: 01 June 2017

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30, 103 and 200 mg/mL for the 300. 1031 and 2000 mg/kg bw groups, respectively.
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: n/a
- Lot/batch no. (if required): n/a
- Purity: n/a

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw (applied from a 200 mg/mL formulation).

DOSAGE PREPARATION (if unusual): The test article was dispersed in purified water. The formulated concentrations were calculated from the selected dose level and the dose volume of 10 mL/kg. All formulations were used within two hours of preparation.
The formulations were either maintained on a magnetic stirrer or the dose containers repeatedly inverted prior to administration, to ensure homogeneity.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Maximum concentration as stipulated in OECD 423.

Doses:

2000, 1031 and 300 mg/kg bw

No. of animals per sex per dose:

6 animals receievd the 300 mg/kg bw dose, groups of 3 animals each received 1031 and 2000 mg/kg bw. Dosing of the group at 1031 mg/kg bw was conducted in error, but does not impact the validity of the study.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Treated rats were observed closely for clinical signs of reaction to treatment. Clinical signs were recorded immediately post-dose, at approximately 15 and 30 minutes post-dose, hourly between 1 and 4 hours post-dose (inclusive), twice daily on Days 2, 3 and 4 and once daily from the fifth to last day of the observation period. Individual records of clinical signs and times of death were maintained for each treated rat. Individual body weights were recorded on Day-1 (day before dosing) and on Days 1, 4, 8 and 15. Decedent carcass weights were also recorded prior to necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Three animals treated at 2000 mg/kg and one animal treated at 1031 mg/kg were found dead one day after dosing. There were no deaths amongst animals treated at 300 mg/kg.

Clinical signs:

other: No clinical signs were seen in animals treated at 300 mg/kg. Common clinical signs seen in animals treated at 1031 or 2000 mg/kg were piloerection, hunched posture and decreased activity. The clinical signs developed from 30 minutes after dosing and last

Gross pathology:

No abnormalities were noted at necropsy of animals that died or were killed at the end of the study.

Table 2      Mortality Data

Does Level (mg/kg)	Mortality Ratio	Time of Death after Dosing
300	0/6
1031	1/3	Day 2
2000	3/3	Day 2

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Based on the conditions of this study, the test article was classified as Category 4 in respect of its acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

Executive summary:

OECD 423 (2017) - In an acute oral toxicity study (OECD 423), groups of fasted, 8-9 week old, female Crl:WI(Han) rats were given a single oral dose of Benzenesulfonicacid, 4 -C10 -13 -sec-alkyl derivs., ammonium salts at doses 2000, 1031 and 300 mg/kg bw and observed for 14 days.

 

Oral LD₅₀ Females = >300 - 2000 mg/kg bw.

 

According to the UN GHS classification, the test item can be classified as Category 4 in respect of its acute oral toxicity.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

500 mg/kg bw

Quality of whole database:

ATE cut-off value based on standard data requirements for this endpoint. The required study is of suitable key stdy quality.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
The vapour pressure of the substance was determined to be < 3.2E-02 Pa, which renders inhalation exposure to this substance negligible, negating the need for an acute toxicity study via the inhalation route. This waiver is in accordance with REACH Annex VIII, Section 8.5.2, Column 2 and ECHA Guidance on Information Requirements and Chemical Safety Assessment, Chapter R7.a.

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 July 2018 - 08 August 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study performed under GLP. All relevant validity criteria were met.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

09 October 2017

Deviations:

yes

Remarks:

The deviation was considered to have not affected the integrity or validity of the study

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

RADIOLABELLING INFORMATION (if applicable): N/A
- Radiochemical purity:
- Specific activity:
- Locations of the label:
- Expiration date of radiochemical substance:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL:
- Storage condition of test material: At room temperature in the dark
- Stability under test conditions:
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING: Tested undiluted
- Treatment of test material prior to testing:
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material): N/A

OTHER SPECIFICS: N/A

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: body weight variation did not exceed ±20% of the mean body weight at the start of treatment
- Fasting period before study: not specified
- Housing: individually housed in suspended solid floor polypropylene cages furnished with softwood flake bedding.
- Diet (e.g. ad libitum): free access to food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK)
- Water (e.g. ad libitum): free access to mains drinking water
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 ºC
- Humidity (%): 30 - 70 %
- Air changes (per hr): minimum of 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 : 12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: back and flanks of each animal were clipped free of hair.
- % coverage: Approximately 10% of total body surface
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi‑occluded with a piece of self‑adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- the treated skin and surrounding hair wiped with cotton wool moistened with dimethyl sulfoxide to remove any residual test item.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 2000 mg/kg bw (no correction made for purity)
- Constant volume or concentration used: 1.049 g/mL

Duration of exposure:

24h

Doses:

Limit test - 1000 mg/kg bw
Main 2000 mg/kg bw

No. of animals per sex per dose:

Limit test - 1
Main - 2

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations and mortality checks were conducted at approximately 0.5, 1, 2, 4, 6 hours and subsequently once daily for 14 days. Scoring and criteria were consistent with the Draize Draize. Individual bodyweight were recorded prior to application of the test item on Day -1 (before dosing) and on Days 0, 7 and 14.
- Necropsy of survivors performed: yes

Statistics:

n/a

Preliminary study:

No effect observed

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

There were no deaths

Clinical signs:

other: There were no signs of systemic toxicity.

Gross pathology:

No abnormalities were noted at necropsy.

Table  1. Dermal Reactions at Dose level 2000 mg/kg

Dose Level (mg/kg)

Animal Number and Sex

Observation

Effects Noted After Initiation of Exposure (Days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

2-0 Female

Erythema

2

2

1

0

?e

?e

?e

?e

?e

?e

?e

?e

?e

?e

Edema

1

1

1

1

?od

?od

?od

?od

?od

?od

?od

?od

?od

?od

Other

BlHd

BlHdTh

BlThSp

BlThSp

Sp

SpSk

SpSkSw

SpSkSw

SpSkSw

SpSkSw

SpSk SwSg

SpSk SwSg

SpSk SwSg

SpSkSg

3-0 Female

Erythema

1

2

2

2

?e

?e

?e

?e

?e

?e

?e

?e

?e

?e

Edema

1

1

1

1

?od

?od

?od

?od

?od

?od

?od

?od

?od

?od

Other

ThLf

ThBl

LfBlTh

LfBlTh

SpSw

SpSw

SpSw

SpSw

SpSw

SpSw

SpSw

SpSw

SpSwSk

SsSgSk

3-1 Female

Erythema

1

2

2

2

?e

?e

?e

?e

?e

?e

?e

?e

?e

?e

Edema

1

1

1

1

?od

?od

?od

?od

?od

?od

?od

?od

?od

?od

Other

ThLf

ThBlLf

ThBlLf

ThBlLf

SpSwSk

SpSwSk

SpSkSb

SdSb

SdSb

SdSb

SdSb

SbSpSk

SbSpSkSg

SkSpSg

0     = No reactions                                                        Hd  = Dermal hemorrhage                     

Th  = Thickening of the skin                                           ?e   = Dermal reactions prevented the accurate evaluation of erythema

Lf   = Loss of skin flexibility                                          ?od = Dermal reactions prevented the accurate evaluation of edema

Bl   = Blanching of the skin                                            Ss   = Small superficial scattered scabs                 

Sp  = Hardened brown colored scab

Sw  = Scab undulating

Sk  = Scab cracking

Sg  = Scab lifting to revel glossy skin

Sb  = Scab lifting at edges to reveal dried blood

Sd  = Scab lifting to at edges to reveal further deeper scabbing

Table  2. Individual Body Weight Changes at Dose level 2000 mg/kg

Dose Level (mg/kg)	Animal Number and Sex	Body Weight (g) at Day			Body Weight Change (g) During Week
		0	7	14	1	2
2000	2-0 Female	222	219	231	-3	12
	3-0 Female	229	232	232	3	0
	3-1 Female	232	229	232	-3	3

Table  3. Individual Necropsy Finding at Dose level 2000 mg/kg

Dose Level (mg/kg)	Animal Number and Sex	Time of Death	Macroscopic Observations
2000	2-0 Female	Killed Day 14	No abnormalities detected
	3-0 Female	Killed Day 14	No abnormalities detected
	3-1 Female	Killed Day 14	No abnormalities detected

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in female Wistar (RccHan:WIST) strain rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Executive summary:

A study was performed according to OECD TG 402; Acute Toxicity (Dermal) and in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar (RccHan:WIST)  strain rat.  Initial test was conducted with semi-occlusive application of undiluted test item at 1000 mg/kg bw to the clipped skin of one animinal. this is then followed by application of a single dose of the test item at a dose level of 2000 mg/kg body weight for a duration of 24 hour under semi-occluded condition and sequent observation for 14 days.

There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. Two animals treated at a dose level of 2000 mg/kg showed body weight loss with the remaining animal showing an expected gain in body weight during the first week. Two animals treated at a dose level of 2000 mg/kg showed expected gains in body weight with the remaining animal showing no gain in body weight during the second week. The animal treated at a dose level of 1000 mg/kg showed an expected gain in body weight during the observation period.. Signs of dermal irritation noted included very slight to well defined erythema and very slight edema, scattered areas of blanching over the treatment site, scattered areas of dermal hemorrhage over the treatment site, thickening of the skin, hardened light brown colored scabs, scab cracking, scab undulating, scab lifting to reveal glossy skin, scab lifting edge to reveal dried blood, scab lifting at edges to reveal further deeper scabbing, small superficial scattered scabs, glossy skin, loss of skin flexibility and loss of skin elasticity.  The dermal reactions prevented the accurate evaluation of erythema and edema at several time points.

Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in female Wistar (RccHan:WIST) strain rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The endpoint is concluded based on a single key study with a Klimisch rating of 1. No effects were observed up to the limit dose of 2000 mg/kg bw/day

Additional information

Acute Toxicity: Oral

OECD 423 (2017) - In an acute oral toxicity study (OECD 423) conducted on the registered substance, groups of fasted, 8-9 week old, female Crl:WI(Han) rats were given a single oral dose of Benzenesulfonicacid, 4 -C10 -13 -sec-alkyl derivs., ammonium salts at doses 2000 and 300 mg/kg bw and observed for 14 days.  Oral LD50 Females = >300 - 2000 mg/kg bw.  According to the UN GHS classification, the test item can be classified as Category 4 in respect of its acute oral toxicity.

Acute Toxicity: Inhalation

The vapour pressure of the substance was determined to be < 3.2E-02 Pa, which renders inhalation exposure to this substance negligible, negating the need for an acute toxicity study via the inhalation route.  This waiver is in accordance with REACH Annex VIII, Section 8.5.2, Column 2 and ECHA Guidance on Information Requirements and Chemical Safety Assessment, Chapter R7.a..

Acute Toxicity: Dermal

OECD 402 (2018) - In an acute dermal toxicity study (OECD 402) conducted on the registered substance, groups of fasted, 8-9 week old, female Crl:WI(Han) rats were given a single dose of undiluted test item, Benzenesulfonicacid, 4 -C10 -13 -sec-alkyl derivs., ammonium salts at doses 2000 mg/kg bw and observed for 14 days.  Oral LD50 Females =  >2000 mg/kg bw.  According to the UN GHS classification, the test item does not meet the classified with respect to acute dermal toxicity.

Justification for classification or non-classification

Acute oral toxicity - Oral LD50 Females = >300 - 2000 mg/kg bw. According to the UN GHS classification, the test item can be classified as Category 4 in respect of its acute oral toxicity.

The substance does not meet the classification criteria for acute dermal or acute inhalation toxicity.