Aluminum chloride hydroxide sulfate

Endpoint summary

Description of key information

-       Oral LD50 : Weight of evidence approach

LD50 = 11800 mg/kg bw (OECD 401, WOE, Rel.3, in rats)

300 < LD50 < 2000 mg/kg bw but not classified (OECD 423, WOE, rel.1; read-across on aluminium chloride, basic)

2000 < LD50 < 5000 mg/kg bw (OECD 401; WOE, rel.2; read-across on aluminium sulphate, hydrate)

Conclusion: 2000 < Oral LD50 < 5000 mg/kg bw as a worst-case

-       Dermal LD50> 2000 mg/kg bw (OECD 402, K, Rel.1, limit test in rats)

-       Inhalation LC50 > 5 mg/L air (= 5000 mg/m3) (OECD 403, K, Rel.1, limit test/nose only in rats)

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:

experimental study

Adequacy of study:

weight of evidence

Study period:

Somewhere during 1978

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other:

Remarks:

Rel. 3 because information on substance identity and composition is lacking. Batch No. is given and method is comparable to guideline 401. If substance identity and composition is known reliability can turn into 2 (limited reported study, pre-GLP).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Principles of method if other than guideline:

Principles other than guideline:
- no necropsy of survivors
- very high concentrations tested

GLP compliance:

no

Remarks:

pre-GLP

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: IFFA CREDO
- Age at study initiation: no info
- Weight at study initiation: 145 - 165 g
- Fasting period before study: approximately 19 hours
- Housing: 5 per cage
- Diet (e.g. ad libitum): IFFARAT
- Water (e.g. ad libitum): water
- Acclimation period: no info


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 50 ± 10
- Air changes (per hr): 8
- Photoperiod (hrs dark / hrs light): no info


IN-LIFE DATES: No info

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: not applicable


MAXIMUM DOSE VOLUME APPLIED: 14 ml/kg


DOSAGE PREPARATION (if unusual): not applicable


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not applicable

Doses:

Main study: 7.2 g/kg, 9.6 g/kg, 12 g/kg, 14.4 g/kg, 16.8 g/kg
Preliminary study: 6 g/kg, 12 g/kg, 24 g/kg

No. of animals per sex per dose:

Main study: 5
Preliminary study: 2

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations after 15 minutes, 1, 2 and 6 hours and every day during 14 days and autopsy of the animals which died. Weighing on Day 0, 1, 2, 4, 7 and 14.
- Necropsy of survivors performed: no
- Other examinations performed: clinical signs, body weight

Statistics:

Calculation of the LD50 by 3 methods:
- Probit method (Bliss - 1935)
- The method of Litchfield and Wilcoxon (1949)
- The method of arcsinus
1. Bliss, C.I. The comparison of dosage-mortality data - Ann. Appl. Biol. 22, 307-333, 1935
Bliss, C.I. The calculation of the dosage-mortality curve - Ann. Appl. Biol. - 22, 134-167, 1935
Bliss, C.I. The determination of the dosage-mortality curve from small numbers - Quart. J; Pham. and Pharmacol., 11 192-216,1935
2. Litchfield J.T., Wilcoxon J.R. and F.: A simplified method of evaluating dose-effect experiments. The Journal of Pharmacology and Experimental Therapeutics, 96, 2, 99-113, 1949

Preliminary study:

Mortality:
- Dose group 6 g/kg: no mortality
- Dose group 12 g/kg: 2 female animals died
- Dose group 24 g/kg: 2 male and 2 female animals died
Clinical signs:
- Dose group 24 g/kg: at 1 hour observation zero spontaneous activity, absence of reflex overturning, piloerection.
- Dose group 12 g/kg: at 1 hour observation apathy, piloerection, motor difficulties and rapid breathing. After 1 day: Prostration and piloerection among the survivors.
- Dose group 6 g/kg: Apathy and piloerection.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

11 800 mg/kg bw

95% CL:

> 10 460 - < 13 360

Remarks on result:

other: Based on Litchfield and Wilcoxon

Mortality:

- Dose group 7.2 g/kg: no mortality
- Dose group 9.6 g/kg: no mortality
- Dose group 12 g/kg: 2 males and 4 females died
- Dose group 14.4 g/kg: 5 males and 4 females died
- Dose group 16.8 g/kg: 5 males and 5 females died

Clinical signs:

- Dose group 7.2 g/kg: Slowdown in spontaneous activity (2 h); at 6 h also piloerection; after 1 day normal behaviour.
- Dose group 9.6 g/kg: Same symptoms as the previous dose.
- Dose group 12 g/kg: low spontaneous activity (2 h); low activity, depression and piloerection (6 h); low activity, depression among all the females and one male, piloerection, one female had blood in her urine, one male had bladder haemorrhagic fever (Day 1); persistence of a slight piloerection among the survivors on Day 3; the surviving animals showed normal behaviour on Day 4.
- Dose groups 14.4 g/kg and 16.8 g/kg: slowdown in spontaneous activity (1 h); also depression, piloerection and staggering gait after 6 hours; on Day 1 the remaining animal died.

Body weight:

Bodyweight gains per dose group were recorded for all surviving rats.

Gross pathology:

- Dose group 12 g/kg: the autopsy by two males and one female showed bleeding of their stomachs and a contracted jejunum.
- Dose groups 14.4 and 16.8 g/kg: no info

Other findings:

No.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The acute oral median lethal dose to rats of Chlorosulfate basique d’aluminium (WAC®) was found to be 11800 mg/kg bodyweight. Based on the assumption that approximately 20% of this test material consists of salts the acute oral median lethal dose to rats of the salts is 2360 mg/kg bodyweight. According to OECD-GHS the salts of chlorosulfate basique d’aluminium is classified as category 5.

Executive summary:

An acute oral toxicity was performed. The method of the study shows similarities to OECD Guideline 401 (Acute Oral Toxicity), and is however not performed according to GLP standards. The test material, Chlorosulfate basique d’aluminium (WAC®) was evaluated for its acute oral toxicity potential in rats when administered as gavage doses at levels of 7200, 9600, 12000, 14400, 16800 mg/kg bw to 5 males and 5 females per dose group.

At 12000 mg/kg 6 animals died, at 14400 mg/kg 9 animals died and at 16800 mg/kg 10 animals died.

Clinical signs of toxicity included pilo-erection and depression; recovery in surviving animals was complete not later than Day 4.

Gross pathologic examination for the deceased animals revealed bleeding of the stomachs.

 

Acute oral LD50(test material) = 11800 mg/kg bw.

Based on the assumption that approximately 20% of this test material consists of salts, acute oral LD50(salts) = 2360 mg/kg bw

 

Under the test conditions, the substance is not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and classified in category 5 according to the GHS.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

30 March 2010 to 22 April 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 11-12 weeks old) were selected.
- Weight at study initiation: 176-223 gram. Body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water (e.g. ad libitum): free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.
- Other details:
A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health.
Results of analysis for diet (nutrients and contaminants), sawdust, paper and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9 – 21.7ºC
- Humidity (%): 34 - 54% Temporary deviations from the minimum level of relative humidity of 40% occurred, but laboratory historical data do not indicate an effect of these deviations.
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN LIFE DATES: From: 30 March 2010 to 22 April 2010

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Remarks:

Correction was made for the purity of the test substance, which was set at 35% for calculations in consultation with the Study Monitor.

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 4.19 mL/kg for 2000 mg/kg and 0.629 mL/kg for 300 mg/kg.

DOSAGE PREPARATION (if unusual):
The test substance was dosed undiluted as delivered by the sponsor. Dose levels were corrected for the purity of the test substance, which was set at 35% for calculations in consultation with the Study Monitor. Dose volumes were calculated as: Dose level (g/kg) / density (g/mL).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Not indicated

Doses:

2000 mg/kg and 300 mg/kg. Single dosage, on Day 1.
Dose levels were corrected for the purity of the test substance, which was set at 35% for calculations.

No. of animals per sex per dose:

2000 mg/kg: 3 females
300 mg/kg: 6 females (2 groups of 3 animals were treated in a stepwise fashion)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (Day 1 - Day 15)
- Frequency of observations and weighing:Clinical signs at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15 (see 'Any other information on materials and methods inc. tables). Body weights on Days 1 (pre-administration), 8 and 15 and at death.
- Necropsy of survivors performed: yes (see 'Any other information on materials and methods inc. tables)
- Other examinations performed: Mortality/viability was observed twice daily. The time of death was recorded as precisely as possible.

Statistics:

No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Preliminary study:

Not applicable.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

other: Dose levels in report were corrected for the purity of the test substance, which was set at 35% for calculations

Sex:

female

Dose descriptor:

other: LD50 cut-off value

Effect level:

500 mg/kg bw

Based on:

other: Dose levels in report were corrected for the purity of the test substance, which was set at 35% for calculations

Remarks on result:

other: According to the OECD 423 test guideline

Mortality:

The incidence of mortality was as follows:
- At 2000 mg/kg: 3/3
- At 300 mg/kg: 1/3

The decedents were found dead within 24 hours post-treatment.

Clinical signs:

Clinical signs observed during the study period were as follows:
- At 2000 mg/kg: Lethargy, hunched posture, piloerection and ptosis.
- At 300 mg/kg: Lethargy, hunched posture, uncoordinated movements, piloerection and/or ptosis.

The surviving animals had fully recovered from the symptoms between Days 4 and 6.

Body weight:

The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

Animals that were found dead at 2000 mg/kg and 300 mg/kg showed a combination of the following findings at macroscopic post mortem examination: beginning or advanced autolysis, black-brown discolouration of the glandular mucosa of the stomach, gray-white, black-brown or reddish foci on the glandular mucosa of the stomach, hardened glandular mucosa of the stomach, dark red foci on the forestomach, distention of the stomach with gas, greenish and dark red foci on lungs, hardened lungs, dark red discolouration of the pancreas, enlargement of the spleen, reddish discolouration of the wall of the duodenum, dark red or black-brown foci on the thymus, yellowish foci on the uterus and yellowish discolouration of uterine adipose tissue in the abdominal cavity.

No macroscopic abnormalities were observed for animals at 300 mg/kg that survived until termination.

Other findings:

Not appplicable.

None

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions of this study, the oral LD50 value of 202029/A in Wistar rats was established to be within the range of 300-2000 mg/kg bw as pure equivalent. However, it was assumed that the mortality observed was due to corrosivity in the gastrointestinal tractus as this was severe local effects and not systemic effects. Therefore, the test substance is not classified for the acute oral toxicity according to the Regulation (EC) N° 1272/2008 (CLP) criteria and to the GHS.

Executive summary:

In an acute oral toxicity study performed according to OECD Guideline 423 and in compliance with GLP, the test substance (202029/A) was administered by oral gavage to three female Wistar rats at 2000 mg/kg bw. In a stepwise procedure additional groups of females were dosed at 300 mg/kg bw. Animals were then observed for mortality, clinical signs for 14 days and were all sacrificed for macroscopic examination. Dose levels were corrected for the purity of the test substance, which was set a 35% for calculation with the Study Monitor.

All animals were dead at 2000 mg/kg bw. One animal was found dead at 300 mg/kg bw. Lethargy, hunched posture, uncoordinated movements, piloerection and/or ptosis. The surviving animals had fully recovered from the symptoms between Days 4 and 6. The body weight gain shown by the surviving animals over the study period was considered to be normal.

Animals that were found dead at 2000 mg/kg bw and 300 mg/kg bw showed a combination of the following findings at macroscopic post mortem examination: beginning or advanced autolysis, black-brown discolouration of the glandular mucosa of the stomach, gray-white, black-brown or reddish foci on the glandular mucosa of the stomach, hardened glandular mucosa of the stomach, dark red foci on the forestomach, distention of the stomach with gas, greenish and dark red foci on lungs, hardened lungs, dark red discolouration of the pancreas, enlargement of the spleen, reddish discolouration of the wall of the duodenum, dark red or black-brown foci on the thymus, yellowish foci on the uterus and yellowish discolouration of uterine adipose tissue in the abdominal cavity. No macroscopic abnormalities were observed for animals at 300 mg/kg that survived until termination.

According to the protocol the animals were administered the substance as such (solution at 35%). Taking into account the substance density of 1.36 g/mL, it means that doses of 5698 mg/kg bw (4.19 mL/kg bw) and 855 mg/kg bw  (i.e 0.629 mL/kg bw) of solution at 35% were administered to the animals. However, the LD50 was expressed as equivalent pure substance and it was within the range of 300 to 2000 mg/kg bw, e.g. the pure substance was classified in cat 4. If the LD50 of the solution at 35% was considered, i.e.between 855 mg/kg bw to 5698 mg/kg bw, the lowest LD50 limit would drive to a classification acute tox. cat 4 as it ranged within 300 to 2000 mg/kg bw of acute tox. cat. 4. However, the mortality observed was due to corrosivity in the gastro-intestinal tractus, as this was severe local effects and not systemic effects. The substance was already classified in cat. 1 for the eyes (see section 7.3.1) . This could be linked to the pH value of 1 for the substance which was tested for the acute oral toxicity and the irritation to eyes.

Under the test conditions of this study, the oral LD50 value of 202029/A in Wistar rats was established to be within the range of 300-2000 mg/kg bw as pure equivalent. However, it was assumed that the mortality observed was due to corrosivity in the gastrointestinal tractus as this was severe local effects and not systemic effects. Therefore, the test substance is not classified for the acute oral toxicity according to the Regulation (EC) N° 1272/2008 (CLP) criteria and to the GHS.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Only raw data were reported, details on test materials are missing. Guideline 401 is followed with deviations.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

: not always a 14 day observation period.

Principles of method if other than guideline:

Not applicable.

GLP compliance:

no

Remarks:

pre-GLP

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:

No data

Route of administration:

oral: unspecified

Vehicle:

other: water or Tween 80 with water

Details on oral exposure:

No data

Doses:

- 2000 mg/kg
- 5000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (not always a 14 day observation period.)
- Frequency of observations and weighing: observations: daily
- Necropsy of survivors performed: yes

Statistics:

No data

Preliminary study:

Not relevant

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 - < 5 000 mg/kg bw

Sex:

male

Dose descriptor:

LD50

Effect level:

< 5 000 mg/kg bw

Mortality:

- No animals died following treatment or during the observation period at the 2000 mg/kg dosage.
- All animals died during the observation period at the 5000 mg/kg dosage. Deaths occurred on the first or second day.

Clinical signs:

- All rats generally exhibited normal appearance and behaviour following treatment and during the observation period at the 2000 mg/kg dosage.
- Clinical observation data at the 5000 mg/kg dosage: The most frequently observed changes included depression, ruffled fur and nearly closed eyes or glossy eyes.

Body weight:

No data

Gross pathology:

- Animals in the 2000 mg/kg group with water as a vehicle showed no gross abnormalities.
- The observed changes of the gross necropsies performed on animals which died during the observation period included very red lung edges and swollen stomach.

Other findings:

See table below (Table 7.2.1/1)

Table 7.2.1/1: Results on deaths, clinical and necropsy observations:

Date	Sex	Dose (mg/kg)	Vehicle	# deaths	Total # animals	Clinical observations	Necropsy observations
8July1976	M	5000	Tween 80 + H2O	5	5	Day 1: - Restless - Ruffled fur - Lying in one place - Occasionally readjust positions - React to touch + sound Day 2: - Remaining rat sits in one place, wobbling slightly - Head rests on floor - Eyes sunken + glossy - Fur ruffled	Day 2: - Lung edges very red - Stomach swollen; content reddish - Yellow liquid filled mouth during necropsy
18June1976	F	2000	Tween 80 + H2O	0	5	Appear normal
13July1976	F	5000	Tween 80 + H2O	5	5	Day 1: - Mild to severe depression - Slower reacting to sound + touch - Ruffled fur - Walking uncoordinated - Eyes partially closed - Huddled in a pile - Mild lacrimation - 1 animal died Day 2: - Other animals died
03August1976	F	2000	H2O	0	5	Appear normal	No gross abnormalities

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the test conditions of this study, the acute oral median lethal dose to rats of Aluminum sulphate, hydrate was found to be greater than 2000 mg/kg body weight and less than 5000 mg/kg bw. Based on these results, the registered substance is not classified according to the Regulation (EC) N° 1272-2008 (CLP) and classified in classified in category 5 according to the GHS.

Executive summary:

In an acute oral toxicity study performed with some equivalence to OECD Guideline 401 but not in compliance with GLP, groups (5/sex/dose) of Sprague Dawley rats were given Aluminum sulphate, hydrate at 2000 mg/kg in water or in Tween 80/water to females and 5000 mg/kg in Tween 80/water to males and females.

Animals were then observed for mortality, clinical signs for 14 days in most cases and were all sacrificed for macroscopic examination.

No mortality occurred in all ten females dosed at the 2000 mg/kg level. No clinical changes were observed generally at this dose level. All ten animals at 5000 mg/kg bw died with deaths occurring on the first or second day. Clinical signs of toxicity included depression and ruffled fur. Necropsy findings showed very red lung edges and/or swollen stomach.

 

The acute oral LD50 for Aluminum sulphate, hydrate was found to be greater than 2000 mg/kg bodyweight and less than 5000 mg/kg bw for females. For males the LD50 is less than 5000 mg/kg. Based on these results, the registered substance was not classified according to the Regulation (EC) N° 1272-2008 (CLP) and classified in category 5 according to the GHS.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Three studies were considered in a weight of evidence approach to assess the acute oral toxicity of the registered substance.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 March 2010 - 05 April 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

This study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: - JMAFF, 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals were selected (approximately 10 weeks old).
- Weight at study initiation: body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: No.
- Housing:
Before exposure
Group housing of five animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
After exposure
Group housing as described above, except that a paper sheet was introduced into the cage covering the bedding and cage enrichment to prevent suffocation in case of bad health condition. At the end of the Day of exposure the paper sheet was removed.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance.
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance.
- Acclimation period: At least 5 days before start of treatment under laboratory conditions.
- Health inspection: A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health

Animals 4 and 5 were replaced by spare animals, 30 minutes after start of exposure, and exposed immediately after restraining. The animals were not acclimatised for at least 15 minutes. The animals were replaced since the original animals were found dead during exposure, possibly caused by stress due to restraining. Since the exposure was ongoing, it was not possible to acclimatise the animals to restraining. It was considered that this event did not affect the exposure results for these animals.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.8 – 21.7°C
- Humidity (%): 42 - 60%
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: From: 22 March 2010 To: 05 April 2010

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Remarks:

unchanged

Mass median aerodynamic diameter (MMAD):

ca. 4 - ca. 5 mm

Geometric standard deviation (GSD):

ca. 1.7

Remark on MMAD/GSD:

The droplet size distribution was representatively characterized twice during the exposure period. The samples were drawn (2 L/min) from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber (appendix 1, figures 1 and 2). The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (SKC 225-713, Fiber glass, SKC Omega Specialty Division, Chelmsford, MA, USA) and a fiber glass back-up filter (-290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test substance collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined.
 
The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. The MMAD was 4.0 and 5.0mm and the gsd was 1.7 at both occasions.

Details on inhalation exposure:

The design of the exposure chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of three animal sections with eight animal ports each. Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained at a slight negative pressure.

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

4 h

Concentrations:

1.6 mg/L, corresponding to 6.1 mg/L of the neat test substance (correction factor 3.7 for the evaporable water content).

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

202028/A was administered as an aerosol by inhalation for 4 hours to one group of five male and five female Wistar rats. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (day 15).

Statistics:

No data.

Preliminary study:

Not applicable.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

5 mg/L air

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: Clinical signs observed among most animals after exposure included lethargy, hunched posture and/or piloerection on Days 1 and/or 2. No clinical signs were observed during exposure

Body weight:

Mean body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

None.

None

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the inhalatory LC50, 4h value of the substance 202028/A (aerosol) in rats was considered to be higher than 5 mg/L. Therefore, no classification is required according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Executive summary:

An acute toxicity study by inhalation was performed according to the OECD guideline No. 403 and in compliance with GLP. 202028/A was administered as an aerosol by inhalation for 4 hours to one group of five male and five female Wistar rats. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (day 15).

The mean actual dry test substance concentration was 1.6 ± 0.5 mg/L, corresponding to 6.1 ± 1.8 mg/L of the neat test substance (correction factor 3.7 for the evaporable water content). The mean nominal concentration was 9.4 mg/L resulting in a generation efficiency (ratio of mean actual concentration and nominal concentration of neat test substance) of 65%.

The concentration measurements conducted equally distributed over time showed that the substance was sufficiently stable.

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. The MMAD was 4.0 and 5.0 μm and the gsd was 1.7 at both occasions.

No mortality occurred.

Clinical signs observed among most animals after exposure included lethargy, hunched posture and/or piloerection on Days 1 and/or 2. No clinical signs were observed during exposure.

The body weight gain shown by the animals over the study period was considered to be normal.

No abnormalities were found at macroscopic post mortem examination of the animals.

Agglomeration of aerosol particles at this high concentration of 5 mg/L may have resulted in one MMAD value to exceed the recommended range of 1 - 4 μm (values obtained were 4.0 and 5.0 μm). Since the values were close to or at the top of the range, it was considered that sufficient test substance deposition in the lower respiratory tract occurred during the exposure in order to give a reliable outcome.

LC50(4h, aerosol) > 5 mg/L air

Under the test conditions of this study, the inhalatory LC50, 4h value of 202028/A (aerosol) in rats was considered to exceed 5 mg/L. Based on these results, no classification is required according to Regulation (EC) No 1272/2008 (CLP) and to the GHS.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 000 mg/m³

Quality of whole database:

The key study is GLP compliant and of high quality (Klimisch score = 1)

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:

11 May 2010 - 25 May 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

This study has been performed according to OECD and EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines (2000), including the most recent revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Crl:WI (Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 10 weeks old) were selected
- Weight at study initiation: Males: 281 - 310 gram; Females: 196 - 238 gram. Body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: No.
- Housing: Individually housed in labeled Macrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom). During the acclimatization period the animals were group housed in Macrolon cages (MIV type).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: Acclimatization period was at least 5 days before start of treatment under laboratory conditions.
- Health inspection: A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from any abnormality.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6 – 21.4ºC
- Humidity (%): 40 – 62%
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: From: 11 May 2010 To: 25 May 2010

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
The test substance was applied in an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The test substance was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
After 24 hours of application, dressings were removed and the skin cleaned of residual test substance using tap water.

TEST MATERIAL
2000 mg/kg (1.682 mL/kg) body weight. The purity of active ingredient (water free) ≥ 69.1 % (Al content 4.4%).
Dose volume calculated as dose level (g/kg) / density (g/mL).

Duration of exposure:

24 hrs

Doses:

2000 mg/kg (1.682 mL/kg) body weight. The purity of active ingredient (water free) ≥ 69.1 % (Al content 4.4%).

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Observations
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
Necropsy: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

Not applicable.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: The purity of active ingredient (water free) ≥ 69.1 % (Al content 4.4%).

Mortality:

No mortality occurred.

Clinical signs:

Two females showed chromodacryorrhoea (snout) and one male showed piloerection on Day 1.

Body weight:

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, 202028/A (purity of active ingredient (water free) ≥ 69.1 %; Al content 4.4%) is not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS as the estimated dermal LD50 is higher than 2000 mg/kg bw and as no toxicological effects is expected between 2000 and 5000 mg/kg bw.

Executive summary:

In an acute dermal toxicity study (limit test) performed according to the OECD guideline No 402 and in compliance with GLP, a group of Wistar rats (5/sex) were given a single dermal application of 202028/A at 2000 mg/kg bw. The test substance 202028/A (purity of active ingredient (water free) ≥ 69.1 %; Al content 4.4%) was applied topically to the shaved skin area of the animals and covered with occlusive binding for 24h. After the exposure period, the dressing was removed and the skin was cleaned of residual test substance using tap water.

Animals were observed for toxicity, clinical signs and body weight for 15 days and then necropsied for macroscopic observations.

 

No mortality occured.

Two females showed chromodacryorrhoa (snout) and one male showed piloerection on Day 1.

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

No macroscopic abnormalities were found during the necropsy.

 

Dermal LD50 rabbit > 2000 mg/kg bw

 

Under the test conditions, 202028/A (purity of active ingredient (water free) ≥ 69.1 %; Al content 4.4%) is not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS as the estimated dermal LD50 is higher than 2000 mg/kg bw and as no toxicological effects is expected between 2000 and 5000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The key study is GLP compliant and of high quality (Klimisch score = 1)

Additional information

Acute toxicity: oral

Three studies were considered in a weight of evidence approach to assess the acute oral toxicity of the registered substance. One study was perfomed on the registered substance and the remaining studies on two analogue substances.

Study 1: Coquet, 1978

The method of the first study shows similarities to OECD Guideline 401 (Acute Oral Toxicity), and is however not performed according to GLP standards. The test material, Chlorosulfate basique d’aluminium (WAC®) was evaluated for its acute oral toxicity potential in rats when administered as gavage doses at levels of 7200, 9600, 12000, 14400, 16800 mg/kg bw to 5 males and 5 females per dose group.

At 12000 mg/kg 6 animals died, at 14400 mg/kg 9 animals died and at 16800 mg/kg 10 animals died.

Clinical signs of toxicity included pilo-erection and depression; recovery in surviving animals was complete not later than Day 4.

Gross pathologic examination for the deceased animals revealed bleeding of the stomachs.

Acute oral LD50 (test material) = 11800 mg/kg bw.

Study 2: Stitzinger, 2010

The second acute oral toxicity study was performed according to OECD Guideline 423 and in compliance with GLP. The test substance (202029/A) was administered as supplied by oral gavage to three female Wistar rats at 2000 mg/kg bw. In a stepwise procedure additional groups of females were dosed at 300 mg/kg bw. Animals were then observed for mortality, clinical signs for 14 days and were all sacrificed for macroscopic examination. Dose levels were corrected for the purity of the test substance, which was set a 35% for calculation with the Study Monitor.

All animals were dead at 2000 mg/kg bw. One animal was found dead at 300 mg/kg bw. Lethargy, hunched posture, uncoordinated movements, piloerection and/or ptosis. The body weight gain shown by the surviving animals over the study period was considered to be normal.

Under the test conditions of this study, the oral LD50 value of 202029/A in Wistar rats was established to be within the range of 300-2000 mg/kg bw as pure equivalent. However, it was assumed that the mortality observed was due to corrosivity in the gastrointestinal tractus as this was severe local effects and not systemic effects.

Study 3: Scholler, 1976

The third acute oral toxicity study was performed with some equivalence to OECD Guideline 401 but in pre-GLP. Groups (5/sex/dose) of Sprague Dawley rats were given Aluminum sulphate, hydrate at 2000 mg/kg in water or in Tween 80/water to females and 5000 mg/kg in Tween 80/water to males and females.

No mortality and nor clinical signs were observed at 2000 mg/kg bw. All ten animals at 5000 mg/kg bw died with deaths occurring on the first or second day. Clinical signs of toxicity included depression and ruffled fur. Necropsy findings showed very red lung edges and/or swollen stomach.

2000 < Acute oral LD50 < 5000 mg/kg bw

Conclusion: 2000 < Acute oral LD50 < 5000 mg/kg bw as a worst-case

Acute toxicity: dermal

A key study was identified (Stitzinger, 2010). In this acute dermal toxicity study (limit test) performed according to the OECD Guideline No 402 and in compliance qwith GLP, a group of Wistar rats (5/sex) were given a single dermal application of 202028/A at 2000 mg/kg bw. The test substance 202028/A (purity of active ingredient (water free) ≥ 69.1 %; Al content 4.4%) was applied topically to the shaved skin area of the animals and covered with occlusive binding for 24h. After the exposure period, the dressing was removed and the skin was cleaned of residual test substance using tap water.

Animals were observed for toxicity, clinical signs and body weight for 15 days and then necropsied for macroscopic observations.

 

No mortality occured.

Two females showed chromodacryorrhoa (snout) and one male showed piloerection on Day 1.

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

No macroscopic abnormalities were found during the necropsy.

 

Dermal LD50 rabbit > 2000 mg/kg bw

Acute toxicity: inhalation

A key study was identified (van Huygevoort, 2010). This acute toxicity study by inhalation was performed according to the OECD guideline No. 403 and in compliance with GLP. 202028/A was administered as an aerosol by inhalation for 4 hours to one group of five male and five female Wistar rats. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15 and macroscopic examination was performed after terminal sacrifice (day 15).

The mean nominal concentration was 9.4 mg/L resulting in a generation efficiency (ratio of mean actual concentration and nominal concentration of neat test substance) of 65%.

The concentration measurements conducted equally distributed over time showed that the substance was sufficiently stable.

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. The MMAD was 4.0 and 5.0 μm and the gsd was 1.7 at both occasions.

No mortality occurred. Clinical signs observed among most animals after exposure included lethargy, hunched posture and/or piloerection on Days 1 and/or 2. No clinical signs were observed during exposure. The body weight gain shown by the animals over the study period was considered to be normal.

No abnormalities were found at macroscopic post mortem examination of the animals.

Agglomeration of aerosol particles at this high concentration of 5 mg/L may have resulted in one MMAD value to exceed the recommended range of 1 - 4 μm (values obtained were 4.0 and 5.0 μm). Since the values were close to or at the top of the range, it was considered that sufficient test substance deposition in the lower respiratory tract occurred during the exposure in order to give a reliable outcome.

inhalation LC50(4h, aerosol) > 5 mg/L air

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 (CLP).

 

Self classification:

Acute toxicity via Oral route:

Based on the available data, the substance is:

- not classified according to the Regulation (EC) No. 1272/2008 (CLP) as the LD50 is higher than 2000 mg/kg bw

- classified in category 5 according to the GHS as the LD50 is considered to be higher than 2000 and potentially below 5000 mg/kg bw.

 

Acute toxicity via Dermal route:

Based on the available data, the substance is:

- not classified according to the Regulation (EC) No. 1272/2008 (CLP) as the LD50 is higher than 2000 mg/kg bw

- not classified according to the GHS as the LD50 is expected to be higher than 5000 mg/kg bw.

 

Acute toxicity via Inhalation route:

Based on the available data, the substance is not classified according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS as the LC50 is higher than 5 mg/L air and the GHS criteria are not met.

 

Specific target organ toxicity: single exposure (Oral):

The classification criteria according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS as specific target organ toxicant (STOT) – single exposure, oral are not met since no systemic reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (oral) for a Category 1 classification (C≤ 300 mg/kg bw) and at the guidance value (oral) for a Category 2 classification (2000 mg/kg bw≥C > 300 mg/kg bw). No classification is required.

The criteria for Transient Organ effects (STOT-SE Category 3) according to Annex VI of the Regulation (EC) No. 1272/2008 and to the GHS are not met since narcotic effects were not observed in the acute oral toxicity studies.

 

Specific target organ toxicity: single exposure (Dermal):

The classification criteria according to the Annex VI of the Regulation (EC) No 1272/2008 (CLP) and to the GHS as specific target organ toxicant (STOT) – single exposure, dermal are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (dermal) for a Category 1 classification (C≤ 1000 mg/kg bw) and at the guidance value (dermal) for a Category 2 classification (2000 mg/kg bw≥C > 1000 mg/kg bw). No classification is required.

The criteria for Transient Organ effects (STOT-SE Category 3) according to Annex VI of the Regulation (EC) No. 1272/2008 and to the GHS are not met since narcotic effects were not observed in the acute dermal toxicity studies.

 

Specific target organ toxicity: single exposure (Inhalation):

The classification criteria according to the Annex VI of the Regulation (EC) No 1272/2008 (CLP) and to the GHS as specific target organ toxicant (STOT) – single exposure, inhalation are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (inhalation/dust and mists) for a Category 1 classification (C≤ 1 mg/L) and at the guidance value (inhalation/dust and mists) for a Category 2 classification (5 mg/L ≥ C > 1 mg/L). No classification is required.

The criteria for Transient Organ effects (STOT-SE Category 3) according to Annex VI of the Regulation (EC) No. 1272/2008 and to the GHS are not met since narcotic effects were not observed in the acute inhalation toxicity study.