Benzyl-C12-14-alkyldimethylammonium chlorides

Endpoint summary

Description of key information

Based on the results of the read across studies and in line with its biocides assessment report, the oral and dermal LD50 values for the test substance is considered to be 350 and 2848 mg/kg bw respectively.  

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

From November 07, 1987 to December 03, 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

other: Acute oral toxicity

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

other: undiluted test substance

Doses:

500, 794, 1,260 and 2,000 mg/kg bw

No. of animals per sex per dose:

10

Details on study design:

Dose selection was based upon the results of a range-finding study. Animals, 5 males and 5 females per dose group, were administered the undiluted test substance in a single oral dose by gavage. Animals were observed 1 and 4h after dosing and subsequently once daily for 14 d. Deaths and evidence of overt toxicity were recorded at each observation. Individual body weights were recorded on the d of treatment (Day 0), Days 7 and 14, and at death. All animals were subjected to gross necropsy examination for any macroscopic abnormalities.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 398 mg/kg bw

Based on:

test mat.

95% CL:

>= 298 - <= 542

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

ca. 358 mg/kg bw

Based on:

act. ingr.

95% CL:

>= 247 - <= 519

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

ca. 438 mg/kg bw

Based on:

act. ingr.

95% CL:

>= 288 - <= 665

Gross pathology:

Necropsy of decedents revealed abnormally red lungs, dark livers, haemorrhage and ulceration of the gastric mucosa and congestion of the small intestines. Major abnormalities seen at necropsy of animals killed at termination were white thickened areas of the non-glandular region of the stomach. Scattered white raised areas were also noted.

Interpretation of results:

other: Category 4 based on CLP criteria

Conclusions:

Under the study conditions, the rat LD50 of the read across substance was considered to be 358 mg a.i./kg bw in males, 438 mg a.i./kg bw in f emales and 398 img a.i./kg bw in males and females combined.

Executive summary:

A study was conducted to determine the acute oral toxicity of the read across substance, C12-16 ADBAC (50% active in water) according to OECD Guideline 401, in compliance with GLP. Based on the results of a range finding study, five male and five female rats per dose group weres administered the undiluted read across substance (50% active) by gavage at dose levels of 500, 794, 1260 and 2000 mg/kg bw (i.e., equivalent to 250, 397, 630 and 1000 mg a.i./kg bw). Animals were observed 1 and 4 h after dosing and subsequently once daily for 14 days. Mortality and evidence of overt toxicity were recorded at each observation. Individual body weights were recorded on the day of treatment (Day 0), Days 7 and 14, and at termination. All animals were subjected to gross necropsy examination for any macroscopic abnormalities. One male treated with 1000 mg a.s./kg was found dead six h after dosing; all other deaths were noted one to two days after treatment. Surviving animals made expected bodyweight gains over the study period. Major signs of toxicity observed in both decedent and surviving animals were hunched posture, pilo-erection, decreased respiratory rate, diarrhoea, lethargy and ptosis. Ataxia was noted in animals treated with 397 mg a.s./kg bw and above. There were no survivors following treatment with 630 and 1000 mg a.s./kg. All surviving animals were normal three to four days after treatment.Necropsy of decedents revealed abnormally red lungs, dark livers, haemorrhage and ulceration of the gastric mucosa and congestion of the small intestines. Major abnormalities seen at necropsy of animals killed at termination were white thickened areas of the non-glandular region of the stomach. Scattered white raised areas were also noted. Under the study conditions, the LD50 was determined to be 358 mg a.i./kg bw (95% c.i: 247-519 mg a.s/kg bw) in males, 438 mg a.i./kg bw (95% c.i.: 288 – 665 mg a.s./kg bw) in females and 398 mg a.s./kg bw (95% c.i.: 298 – 542 mg a.s./kg bw) in male and females combined (Jones, 1986). Based on the results of the read across study, similar LD50 value is expected for the test substance.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

June, 1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Basic data given, comparable to scientific principles/standards.

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Principles of method if other than guideline:

Acute oral toxicity was determined by oral administration of the test substance to nine dose groups of male and female rats and subsequent observations of clinical signs and mortality for 14 days. The LD50 was calculated from the mortality data recorded in the study.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animals:
- Age at study initiation: Young adult rats
- Weight at study initiation: 200-300 g
- Fasting period before study: 24h
- Diet: Ad libitum
- Water: Ad libitum

Route of administration:

oral: gavage

Vehicle:

other: Propylene glycol for 1 mL/kg bw and lower doses and undiluted test substance for doses 2 mL/kg bw and above dose levels.

Details on oral exposure:

Vehicle:
- Amount of vehicle: Propylene glycol for 1 mL/kg bw and all lower doses. Higher doses were administered undiluted.

Doses:

0.25, 0.32, 0.40, 0.50, 1.0, 2.0, 4.0, 8.0 and 16.0 mL/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14d
- Frequency of observations: Daily
- Necropsy of survivors performed: No

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

0.43 mL/kg bw

Based on:

test mat.

95% CL:

>= 0.39 - <= 0.47

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

344 mg/kg bw

Based on:

act. ingr.

Mortality:

0/5, 0/5 and 1/5 animals died at 0.25, 0.32 and 0.40 mL/kg bw dose levels respectively. All animals in the higher dose groups (0.50 to 16.00 mL/kg bw) died during the study period.

Clinical signs:

other: All animals dosed at 0.25 to 0.50 mL/kg bw exhibited lethargy and slight to moderate diarrhea. The severity of the symptoms increased proportionately to the dose level received. Surviving animals returned to normal within 5 days of dosing. Animals dosed a

Interpretation of results:

other: Category 4 based on CLP criteria

Conclusions:

Under the conditions of the study, the LD50 of the test substance was 0.43 mL/kg bw (95% c.i.-0.39 - 0.47 mL/kg bw). After correcting for 100% active test substance, the LD50 was determined to be 344 mg a.i./kg bw.

Executive summary:

A study was conducted to determine the acute oral toxicity of the read across substance, C12-16 ADBAC (80% active), in albino rats. The test substance was administered to groups of five fasted male and female albino rats at 0.25, 0.32, 0.40, 0.50, 1.0, 2.0, 4.0, 8.0 or 16.0 mL/kg bw. Propylene glycol was used as vehicle for 1 mL/kg and all lower doses. Doses of 2 mL/kg bw and above were administered as received. Animals were observed for 14 d post-dosing. No post-mortem nor histopathological examinations were performed. All animals dosed with 0.25 to 0.50 mL/kg exhibited lethargy and slight to moderate diarrhea. The severity of the symptoms increased proportionately to the dose level received. Surviving animals returned to normal within 5 days. Animals dosed with 1.0 or 2.0 mL/kg were extremely lethargic. 0/5, 0/5 and 1/5 animals died at 0.25, 0.32 and 0.40 mL/kg bw, respectively. All animals in the higher dose levels died during the study period. Under the conditions of the study, the LD50 of the test substance is considered to be 0.43 mL/kg bw (95% c.i.:0.39 - 0.47 mL/kg bw) in males and females combined. After correcting for 100% active test substance, the LD50 is determined to be 344 mg a.i./kg bw (Wallace, 1975). Based on the results of the read across study, similar LD50 value is expected for the test substance.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

350 mg/kg bw

Quality of whole database:

The information requirement for this tonnage band is sufficiently met with the available data.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

disregarded due to major methodological deficiencies

Study period:

1989

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Principles of method if other than guideline:

Thirteen week subacute inhalation toxicity in the albino rat and golden hamster.

GLP compliance:

not specified

Species:

other: Rats and hamsters

Strain:

other: Albino and Syrian Golden

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Details on study design:

An inhalation toxicity study of an aerosolised hair conditioner containing 0.2% of a 50.0% test substance (effective concentration 0.1%)  was conducted with 12 female albino rats of the CD-strain and 12 Syrian Golden hamsters. Exposures were carried out in 500L dynamic flow inhalation chambers. The animals were exposed to the conditioner (9.9 mg/m3 of air) 5d a week (4 h/d) for 14 consecutive weeks. 

Key result

Dose descriptor:

LC0

Effect level:

> 9.9 mg/m³ air

Mortality:

There were no exposure-related deaths.

Body weight:

There were no significant differences in weight gain.

Gross pathology:

There were no exposure-related gross nor microscopic changes were attributed to test substance inhalation.

Other findings:

- Aerosol concentration in the chamber atmosphere was analysed.
- There were no significant differences in haematological values, and serum chemistry data between experimental and control groups.

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the study conditions, the rat and hamster LC0 of the read across substance was greater than 9.9 mg/m3.

Executive summary:

An inhalation toxicity study was conducted with an aerosolised hair conditioner containing 0.2% of a 50.0% read across substance (effective concentration 0.1%) in 12 female albino rats of the CD-strain and in 12 Syrian Golden hamsters. Exposures were carried out in 500 L dynamic flow inhalation chambers. The animals were exposed to the conditioner (9.9 mg/m3 of air) 5 d a week (4 h/d) for 14 consecutive weeks. Aerosol concentration in the chamber atmosphere was analysed. There were no significant differences in weight gain, haematological values and serum chemistry data between experimental and control groups. There were no exposure-related deaths, and neither gross nor microscopic changes were attributed to read across substance inhalation. Under the study conditions, the rat and hamster LC0 of the read across substance was > 9.9 mg/m3 (CIR, 1989). Based on the results of the read across study, similar LC0 value is expected for the test substance.

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Study period:

From February 21, 1990 to April 30, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

KL2 due to RA

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPP 81-3 (Acute inhalation toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animals:
- Source: Charles River UK Ltd., Margate, UK
- Weight at study initiation: ca. 200 g on the d of exposure
- Housing: 5/sex in polypropylene cages with detachable wire mesh tops and floors
- Diet (e.g. ad libitum): ad lib
- Water (e.g. ad libitum): ad lib
- Acclimation period: at least 5 d

Environmental conditions:
- Temperature (°C): 18-24
- Humidity (%): 35-65

In-life dates: From 21 February, 1990 to 30 April, 1990

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

Generation of test atmosphere / chamber description
- Exposure apparatus: perspex whole body chamber (square section with pyramidal top)
- Exposure chamber volume: ca. 120L
- Method of holding animals in test chamber: wire mesh partitions to provide 10 separate animal compartments
- Source and rate of air: filtered and oil-free compressed air
- Method of conditioning air: dried
- System of generating particulates/aerosols: atomiser
- Method of particle size determination: cascade impaction (Andersen mini-sampler and Marple cascade impactor (model 296)
- Treatment of exhaust air: passage through a collection filter
- Temperature, humidity, pressure in air chamber: temperature measured at 30-min intervals (22-23°C), relative humidity not measured (reason: aqueous solution). However, as dried air was used and the test substance contained only 7.7% water, relative humidity could have been measured. Pressure in air chamber: not indicated. However, as the whole body chamber was placed in a hood, this is not as important.

Test atmosphere
- Brief description of analytical method used:
Five air samples (5 L for groups 2 and 4 (0.34 and 0.24 mg/L), 10L for group 3 (0.17 mg/L) were taken from the chamber during each exposure and the collected material was analysed to determine the concentration of the test substance in the chamber air. Each air sample was withdrawn, at 4 L per min, through a weighed glass fibre filter (Whatman GF/A) mounted in an open face filter holder. The volume of the air sample was measured with a wet-type gas meter. Two further air samples were taken using an Andersen mini-sampler or a series 290 Marple cascade impactor (Model 296), and the collected material was weighed to determine the particle size distribution of the test subtance. The samples were taken at approximately 1.5 and 3.5 h from the start of exposure. The filters from the open face sampler were transferred to extraction columns and compacted with a glass road. The test substance was eluted with five 2 mL portions of methanol into a 20 mL volumetric flask and diluted to volume with methanol.
The filters from the Andersen and Marple samplers were similarly treated t o give a final volume of 5 mL. The stages of the Andersenand Marple samplers were washed off with small amounts of methanol into 5 mL volurnentric flasks. The extracts were diluted with mobile phase to obtain solutions for HPLC-analysis with expected maximum concentrations of the test substance of 150 pg/mL.

- Samples taken from breathing zone: taken from the whole body chamber.

Vehicle: not used

- Test atmosphere (if not tabulated)
Concentrations:
0.34 mg/L (± 6%); nominal: 2.21 mg/L
0.17 mg/L (± 13%); nominal: 0.52 mg/L
0.24 mg/L (± 17%); nominal: 0.88 mg/L

- MMAD (mass median aerodynamic diameter) / GSD (geometric standard deviation):
0.34 mg/L : MMAD 1.9 µm, gsd 3.5
0.17 mg/L : MMAD 1.1 µm, gsd 2.7
0.24 mg/L : MMAD 0.8 µm, gsd 0.8

Analytical verification of test atmosphere concentrations:

yes

Remarks:

HPLC

Duration of exposure:

4 h

Concentrations:

0, 0.17, 0.24 and 0.34 mg/L

No. of animals per sex per dose:

5 per sex per concentration

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 21d
- Frequency of observations and weighing: observations continuously duringe exposure and at least twice daily thereafter; BW daily
- Food and water intake: daily
- Necropsy of survivors performed: yes
- Other examinations performed: lung weight, histopathology of lungs, liver and kidneys

Statistics:

LC50 determination by log probit method of Miller LC and Tainter ML, Proc. Soc. Exp. Bio. Med., 57(2),1944:261-264.

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

0.22 mg/L air (analytical)

95% CL:

>= 0.17 - <= 0.27

Exp. duration:

4 h

Key result

Sex:

female

Dose descriptor:

LC50

Effect level:

0.28 mg/L air (analytical)

95% CL:

>= 0.21 - <= 0.35

Exp. duration:

4 h

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

0.25 mg/L air (analytical)

95% CL:

>= 0.22 - <= 0.28

Exp. duration:

4 h

Mortality:

control group: 0/10
0.17 mg/L: 1 male (1/10)
0.24 mg/L: 3 males and 1 female (4/10)
0.34 mg/L: 5 males and 4 females (9/10)

Clinical signs:

other: During exposure: The signs seen during exposure were considered to be consistent with inhalation of an irritant aerosol. Closing or partial closing of the eyes and exaggerated respiratory movement were seen in all rats exposed to the test subtance. Additi

Body weight:

There were moderate to marked decreases of body weight or reductions in the rate of body weight gain for up to 8d in male rats and for up to 14d in female rats following exposure at 0.17 mg/L or 0.24 mg/L. Subsequently weight gain for rats that survived exposure to the test substance was similar to that of the control rats.

Gross pathology:

The findings for rats that died as a result of exposure to the test substance were typified by congestion of the lungs, fluid in the trachea and gas-filled stomach. Macroscopic abnormalities in a proportion of rats that survived exposure to the test substance were a swollen appearance of the lungs and gas-filled stomachs and intestines.

Other findings:

The food and water consumption for the rat that survived exposure at 0.34 mg/L was variable and reduced. Food consumption was reduced for up to 12 d following exposure to the test substance at concentrations of 0.24 or 0.17 mg/L. The water consumption for these groups was reduced for up to 14d following exposure.

The lung weight to body weight ratio was increased, due to a high lung weight, for most rats that died as a result of exposure to ARMOBLEN 400 for a few decedents and the majority of the surviving rats was increased because of low body weight.

Histopathology
Group 2 (0.34 mg/L test substance)
Decedents
Treatment-related changes were seen in the 5 male and 4 female decedents. The distribution of these lesions was as follows:
focal alveolar wall necrosis in 3 males; diffuse congestion in 3 males and 4 females; eosinophilic material in alveoli in 5 males and 4
females; alveolitis in 2 males and 4 females; focal alveolar oedema in 1 male; perivascular oedema in 4 males.
Survivors
Treatment-related changes were seen in the single female rat surviving to the end of the observation period. These were as
follows: focal alveolitis; focal bronchiolitis; prominent bronchiolar goblet cells.

Group 3 (0.17 mg/L test substance)
Decedents
Treatment-related changes were seen in the single decedent male. These were as follows: diffuse congestion; eosinophilic materialin alveoli; alveolitis; focal alveolar oedema.
Survivors
Treatment-related changes were seen in one of the 4 males and in one of the 5 females surviving to the end of the observation
period. The distribution of these lesions was as follows: focal bronchiolitis in 1male; prominent bronchiolar goblet cells in 1 male;
foreign body giant cells in 1 male; focal alveolar haemorrhage in 1 female. A focus of emphysema was also seen in one surviving
female. The significance of this finding in a single animal is unclear, but possibility that it is related to treatment cannot be excluded.
No abnormalities were detected in 3 males and 3 females surviving to the end of the observation period.

Group 4 (0.24 mg/L test substance)
Decedents
Treatment-related changes were seen in the 3 male decedents and 1 female decedent. The distribution of these lesions was as
follows: diffuse or focal congestion in 2 males and 1 female; eosinophilic material in alveoli in 2 males and 1 female; alveolitis in 1
male and 1 female; focal alveolar oedema in 1 male; focal alveolar haemorrhage in 1 female; perivascular oedema in 1 female.
Survivors
Treatment-related change was seen in one of the two males and one of the 4 females surviving to the end of the observation
period. This lesion was: foreign body giant cells. No abnormalities were detected in 1 male and 3 females surviving to the end of theobservation period.

Comment from the authors
Treatment-related changes were found in the survivors from all groups. It is not possible to predict with certainty the progression of these lesions, but the changes were minimal and of a focal nature and it is possible that they would be resolved in the course of time leaving no permanent lesions.

For result tables, kindly refer to the attached background material section of the IUCLID.

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the study conditions, the 4h-LC50 of the mixture was calculated to be 0.25 mg/L or 250 mg/m3.

Executive summary:

A study was conducted to determine the inhalation acute inhalation toxicity of a mixture of read across substances (77.3% total active) containing cocobenzyldimethylamonium chloride (C12-16 ADBAC; 40% active) and dicocodimethylammonium chloride (di C12-16 DMAC; 37.5% active), according to OECD Guideline 403 and US EPA OPP 81 -3, in compliance with GLP. The experiment was performed in Sprague-Dawley rats. Four groups of ten rats (five males and five females) were given a single, 4 h whole body exposure at concentration levels of 0, 0.17, 0.24 and 0.34 mg/L. The animals were observed for 21 d after exposure and were then culled for gross and histopathological examination of the lungs. Bodyweight, food and water intake and lung weight were also determined. There were no deaths in the control group; one animal (male) died at 0.17 mg/L, four animals died at 0.24 mg/L (3 males, 1 female), and nine animals died at 0.34 mg/L (5 males, 4 females). Clinical signs of toxicity noted were (partial) closing of the eyes and exaggerated respiratory movement during exposure in all test groups, gasping and wetness around the mouth during exposure at 0.34 mg/L. Clinical signs were noted in survivors throughout the 21-day observation period. A decrease in body weight reduced weight gain, and reduced food and water intake were generally seen up to Day 14. Abnormalities noted at necropsy in survivors were increased relative lung weight, swollen appearance of the lungs and gas-filled stomach and intestines. Animals that died showed congestion of the lungs, fluid in the trachea, and gas-filled stomach. Histopathological lung changes in survivors generally consisted of focal alveolitis and bronchiolitis; changes in deceased animals generally consisted of focal alveolar wall necrosis, diffuse congestion, focal alveolar wall oedema and focal alveolar wall haemorrhage. Under the study conditions, the 4 h LC50 of the read across substance was calculated to be 0.25 mg/L or 250 mg/m3 (95% CI: 0.22-0.28 mg/L or 220-280 mg/m3) (Jackson, 1990). Based on the results of the read across study, similar LC50 value is expected for the test substance.

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

study not available with pure substance; study with a mixture showed adverse effect observed; LC50: 250 mg/m³ (or 129 mg a.i./m3)

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

1. In the study, 4 animals/sex/test group were used; however, the guideline recommends 5 animals of one sex in each test group. 2. Abraded and non abraded skin sites were used; however, guideline recommends unabraded skin. 3. Gross necropsy was not perfor

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animals:
- Weight at study initiation: 2.2 – 3.4 kg

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Test site
- Area of exposure: Back of animal (10% of body surface)
- Type of wrap if used: The test sites were covered with gauze and each animal was wrapped in a sleeve after application of the test substance
- Type of test site: Intact and abraded (half of the test animals in each sex, i.e., 2 animals/sex/group had their skin abraded on one side )

Removal of the test substance
- Washing (if done): After removal of the dressing, animals were washed with warm water and dried.
- Time after start of exposure: After 24h

Test material
- Amount(s) applied (volume or weight with unit): 3,4 and 5 mL/kg bw
- Concentration (if solution): Undiluted (as received)
- Constant volume or concentration used: yes

Duration of exposure:

24h

Doses:

3,4 and 5 mL/kg bw

No. of animals per sex per dose:

4

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14d
- Frequency of observations and weighing: The animals were observed for clinical signs, mortality and body weight (at the start of the experiment and at termination (Day 14)).
- Necropsy of survivors performed: No

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

3.56 mL/kg bw

Based on:

test mat.

95% CL:

>= 3.01 - <= 4.2

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 730 mg/kg bw

Based on:

act. ingr.

Mortality:

Mortality observed at each dose levels:
3 mL/kg bw: 1/8
4 mL/kg bw: 6/8
5 mL/kg bw: 7/8

For details please refer to the attachment under 'Attached background material'

Clinical signs:

other: Severe erythema and oedema was observed post dosing at the sites of application for all animals in all groups. In the 3 mL/kg bw dose group (i.e., surviving animals), the erythema was followed by thickening of the skin and eschar formation across the back

Table 1:

Dose levels (ml/kg)	Mortality	Time to mortality
5 0	7/8	1, 2, 7 & 12 days
4.0	6/8	2, 3, 4 & 6 days
3.0	1/8	9 days

Table 2:

Dose levels (ml/kg)	Animal #	Bodyweight (kg)
		Initial	Final
5	58	2.3	2.0
4	57A	2.8	2.7
	61A	2.8	2.6
3	57B	3.0	2.6
	58B	2.7	2.1
	59B	3.2	3.1
	60B	2.7	2.4
	61B	2.6	2.4
	63B	2.3	1.6
	64B	2.6	2.0

Interpretation of results:

other: CLP criteria not met

Conclusions:

Under the conditions of the study, the acute dermal LD50 of the read across substance was found to be 3.56 mL/kg bw (95% c.i.- 3.01 - 4.20 mL/kg bw). After correcting for 80%purity of the active substance, the LD50 was calculated to be 2730 mg/kg bw.

Executive summary:

A study was conducted to determine the acute dermal toxicity of the read across substance, C12-16 ADBAC (80% active in water), according to a method similar to EPA OPPTS 870.1200. The experiment was performed in rabbits. The read across substance was applied to twelve male and twelve female rabbits (4 animals/sex/test group) at dose levels of 3, 4 and 5 mL/kg bw (single application) on the abraded and intact skin of the back. The test sites were covered with gauze and each animal was wrapped in a sleeve after application for a 24 h period. After removal of the dressing, animals were washed with warm water and dried. The animals were observed for clinical signs and mortality for 14 d. Body weights were determined at the start of the experiment and at termination (Day 14). In the study, 1/8, 6/8 and 7/8 animals died at 3, 4 and 5 mL/kg bw, respectively. Decreases in bodyweight were observed in all surviving animals in all treatment groups.Severe erythema and oedema at site of application observed post dosing for all animals in all groups. In the 3 mL/kg dose group (i.e. surviving animals), the erythema was followed by thickening of the skin and eschar formation across the back. Under the conditions of the study, the acute dermal LD50 of the read across substance is considered to be 3.56 mL/kg bw (95% c.i.- 3.01 - 4.20 mL/kg bw). After correcting for 80% purity of the active substance, the LD50 is calculated to be 2730 mg a.i./kg bw (Levenstein, 1977). Based on the results of the read across study, similar LD50 value is expected for the test substance.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 848 mg/kg bw

Quality of whole database:

The information requirement for this tonnage band is sufficiently met with the available data.

Additional information

Oral

Study 1: A study was conducted to determine the acute oral toxicity of the read across substance, C12-16 ADBAC (50% active in water) according to OECD Guideline 401, in compliance with GLP. Based on the results of a range finding study, five male and five female rats per dose group weres administered the undiluted read across substance (50% active) by gavage at dose levels of 500, 794, 1260 and 2000 mg/kg bw (i.e., equivalent to 250, 397, 630 and 1000 mg a.i./kg bw). Animals were observed 1 and 4 h after dosing and subsequently once daily for 14 days. Mortality and evidence of overt toxicity were recorded at each observation. Individual body weights were recorded on the day of treatment (Day 0), Days 7 and 14, and at termination. All animals were subjected to gross necropsy examination for any macroscopic abnormalities. One male treated with 1000 mg a.s./kg was found dead six h after dosing; all other deaths were noted one to two days after treatment. Surviving animals made expected bodyweight gains over the study period. Major signs of toxicity observed in both decedent and surviving animals were hunched posture, pilo-erection, decreased respiratory rate, diarrhoea, lethargy and ptosis. Ataxia was noted in animals treated with 397 mg a.s./kg bw and above. There were no survivors following treatment with 630 and 1000 mg a.s./kg. All surviving animals were normal three to four days after treatment.Necropsy of decedents revealed abnormally red lungs, dark livers, haemorrhage and ulceration of the gastric mucosa and congestion of the small intestines. Major abnormalities seen at necropsy of animals killed at termination were white thickened areas of the non-glandular region of the stomach. Scattered white raised areas were also noted. Under the study conditions, the LD50 of the read across substance was determined to be 358 mg a.i./kg bw (95% c.i: 247-519 mg a.s/kg bw) in males, 438 mg a.i./kg bw (95% c.i.: 288 – 665 mg a.s./kg bw) in females and 398 mg a.s./kg bw (95% c.i.: 298 – 542 mg a.s./kg bw) in male and females combined (Jones, 1986).

Study 2: A study was conducted to determine the acute oral toxicity of the read across substance, C12-16 ADBAC (80% active), in albino rats. The read across substance was administered to groups of five fasted male and female albino rats at 0.25, 0.32, 0.40, 0.50, 1.0, 2.0, 4.0, 8.0 or 16.0 mL/kg bw. Propylene glycol was used as vehicle for 1 mL/kg and all lower doses. Doses of 2 mL/kg bw and above were administered as received. Animals were observed for 14 d post-dosing. No post-mortem nor histopathological examinations were performed. All animals dosed with 0.25 to 0.50 mL/kg exhibited lethargy and slight to moderate diarrhea. The severity of the symptoms increased proportionately to the dose level received. Surviving animals returned to normal within 5 days. Animals dosed with 1.0 or 2.0 mL/kg were extremely lethargic. 0/5, 0/5 and 1/5 animals died at 0.25, 0.32 and 0.40 mL/kg bw, respectively. All animals in the higher dose levels died during the study period. Under the conditions of the study, the LD50 of the read across substance is considered to be 0.43 mL/kg bw (95% c.i.:0.39 - 0.47 mL/kg bw) in males and females combined. After correcting for 100% active read across substance, the LD50 is determined to be 344 mg a.i./kg bw (Wallace, 1975).

Based on the above studies, same effect levels and toxicity potential were concluded in the biocide assessment report available on C12-16 ADBAC by RMS Italy (ECHA biocides assessment report, 2015). The RMS further made a remark about the suitability of the test substance used in the Jones, 1986 study:“Although the test item is different, this result can be considered valid for C12-16-BKC, based on the similar mechanism for oral toxicity shown by QUATS with this alkyl chain length”. Lastly, a mean value of ca. 350 mg/kg bw based on both the studies was taken forward as the key conclusion for the product authorization.

Therefore, in line with the biocides assessment report (ECHA biocides assessment report, 2015) and given that the read across to C12-16 ADBAC can be justified for the test substance based on a category approach, the mean oral LD50 value of 350 mg/kg bw has been considered further for hazard/risk assessment. Overall, the main toxicity following acute oral exposure to the test substance relates to irritation and corrosivity rather than systemic effects.  

Inhalation

Study 1. A study was conducted to determine the inhalation acute inhalation toxicity of a mixture of read across substances (77.3% total active) containing cocobenzyldimethylamonium chloride (C12-16 ADBAC; 40% active) and dicocodimethylammonium chloride (di C12-16 DMAC; 37.5% active), according to OECD Guideline 403 and US EPA OPP 81 -3, in compliance with GLP. The experiment was performed in Sprague-Dawley rats. Four groups of ten rats (five males and five females) were given a single, 4 h whole body exposure at concentration levels of 0, 0.17, 0.24 and 0.34 mg/L. The animals were observed for 21 d after exposure and were then culled for gross and histopathological examination of the lungs. Bodyweight, food and water intake and lung weight were also determined. There were no deaths in the control group; one animal (male) died at 0.17 mg/L, four animals died at 0.24 mg/L (3 males, 1 female), and nine animals died at 0.34 mg/L (5 males, 4 females). Clinical signs of toxicity noted were (partial) closing of the eyes and exaggerated respiratory movement during exposure in all test groups, gasping and wetness around the mouth during exposure at 0.34 mg/L. Clinical signs were noted in survivors throughout the 21-day observation period. A decrease in body weight reduced weight gain, and reduced food and water intake were generally seen up to Day 14. Abnormalities noted at necropsy in survivors were increased relative lung weight, swollen appearance of the lungs and gas-filled stomach and intestines. Animals that died showed congestion of the lungs, fluid in the trachea, and gas-filled stomach. Histopathological lung changes in survivors generally consisted of focal alveolitis and bronchiolitis; changes in deceased animals generally consisted of focal alveolar wall necrosis, diffuse congestion, focal alveolar wall oedema and focal alveolar wall haemorrhage. Under the study conditions, the 4 h LC50 of the read across substance was calculated to be 0.25 mg/L or 250 mg/m3 (95% CI: 0.22-0.28 mg/L or 220-280 mg/m3) (Jackson, 1990).

Study 2. An inhalation toxicity study was conducted with an aerosolised hair conditioner containing 0.2% of a 50.0% read across substance (effective concentration 0.1%) in 12 female albino rats of the CD-strain and in 12 Syrian Golden hamsters. Exposures were carried out in 500 L dynamic flow inhalation chambers. The animals were exposed to the conditioner (9.9 mg/m3 of air) 5 d a week (4 h/d) for 14 consecutive weeks. Aerosol concentration in the chamber atmosphere was analysed. There were no significant differences in weight gain, haematological values and serum chemistry data between experimental and control groups. There were no exposure-related deaths, and neither gross nor microscopic changes were attributed to read across substance inhalation. Under the study conditions, the rat and hamster LC0 of the read across substance was > 9.9 mg/m3 (CIR, 1989).

Except for the above studies with a mixture of quaternary ammonium substances and a formulation with the read across substance, no well conducted with the pure test substance could be identified. However, in accordance with Annex VII, Section 8.5, Column 2, of the REACH regulation, the study does not need to be conducted because the substance is classified as corrosive to the skin. Further, the substance has a low vapour pressure (VP = 0.0058 Pa at 25 °C, based on read across), which is below the cut-off of 0.01 Pa set for defining low volatility substances, as per the ECHA Guidance R.7a (2017). Therefore, due to its solid physical state and low VP, it is unlikely that it will form inhalable dust, mist or fumes when handled and used in solid form. In case inhalable forms of the substance (either pure or in aqueous solutions) are created under particular conditions (e.g., spraying, elevated temperature/pressure), appropriate risk management measures such as closed systems, exhaust ventilation or wearing of respirators are implemented to control exposure. Under such conditions, the risk to humans following inhalation exposure can be considered minimal and further testing involving vertebrate animals may be omitted, in accordance with Annex XI (1.2) of the REACH regulation. 

Dermal

A study was conducted to determine the acute dermal toxicity of the read across substance, C12-16 ADBAC (80% active in water), according to a method similar to EPA OPPTS 870.1200. The experiment was performed in rabbits. The read across substance was applied to twelve male and twelve female rabbits (4 animals/sex/test group) at dose levels of 3, 4 and 5 mL/kg bw (single application) on the abraded and intact skin of the back. The test sites were covered with gauze and each animal was wrapped in a sleeve after application for a 24 h period. After removal of the dressing, animals were washed with warm water and dried. The animals were observed for clinical signs and mortality for 14 d. Body weights were determined at the start of the experiment and at termination (Day 14). In the study, 1/8, 6/8 and 7/8 animals died at 3, 4 and 5 mL/kg bw, respectively. Decreases in bodyweight were observed in all surviving animals in all treatment groups.Severe erythema and oedema at site of application observed post dosing for all animals in all groups. In the 3 mL/kg dose group (i.e. surviving animals), the erythema was followed by thickening of the skin and eschar formation across the back. Under the conditions of the study, the acute dermal LD50 of the read across substance is considered to be 3.56 mL/kg bw (95% c.i.- 3.01 - 4.20 mL/kg bw). After correcting for 80% purity of the active substance, the LD50 is calculated to be 2730 mg a.i./kg bw (Levenstein, 1977).

The biocides assessment report available for C12-16 ADBAC (ECHA biocides assessment report, 2015), had reported the active ingredient corrected LD50 value as 2848 mg a.i./kg bw. Therefore, in line with the biocides assessment report and given that the read across to C12-16 ADBAC can be justified for the test substance based on a category approach, the dermal LD50 value of 2,848 mg/kg bw has been considered further for hazard/risk assessment. Overall, the toxicity is considered secondary to the local tissue damage, rather than the result of percutaneously absorbed material. 

Justification for classification or non-classification

Based on the oral and dermal LD50 values from the read across studies, the test substance C12-14 ADBAC warrants an ‘Acute Tox. 4; H302: harmful if swallowed’ classification for the oral route and 'no classification' for the dermal route according to EU CLP criteria (Regulation EC 1272/2008).

In addition, for the inhalation route, although the test substance is classified to be corrosive (see section 5.3) and this drives its mechanism of action of toxicity, its inherent low vapour pressure prohibits the occurrence of respiratory irritation or corrosion by vapour. Therefore, it does not warrant additional labelling as: EUH071 — ‘Corrosive to the respiratory tract’ according to EU CLP criteria (Regulation EC 1272/2008).  

 

Further, the available data does not show an indication that classification for STOT-SE cat 1 or 2 is indicated. For STOT-SE Cat 3: the test substance, C12-14 ADBAC or other QAS substances are not narcotic.