Aluminum chloride hydroxide sulfate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 04 1986 to September 12 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study was carried out under GLP following OECD guidelines. However, the identity and purity of the compound is not verified by a certificate of analysis, nor is there any phys-chem data for the compound.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

nominal concentrations were prepared by direct addition to test system due to limited water solubility of the test substance

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

no sampling reported

Vehicle:

no

Details on test solutions:

Dilution water
Dilution water was reconstituted according to ISO / DIN 7346/1. The preparation was carried out in a facility, consisting of two steel tub filled with Hostalen lining and a capacity of 1700 liters each. The prepared dilution water was aerated to oxygen saturation. The pH was measured prior to each approach, and was between 8.0 and 8.1.

Preparation of test concentrations
Tanfix AL was examined in addition to the alkaline range (pH 7.9 - 8.1) of the test water used, also in the acidic range at pH 6.0. The substance was mixed with dilution water, stirred for 5 minutes with a magnetic stirrer and stirring with a glass rod. The pH was adjusted with 5N NaOH or 1N HCl. In the alkaline range, a significant precipitation was observed, while in the acidic range, an essentially clear solution existed.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Breeding conditions
Temperature: 22 +/- 1 C
Oxygen: >= 80 % air saturation value
Light duration: 12 hours daily
Stocking density: <= 1 g fish/ liter of water


Feeding: twice daily ad libitum
Food: Tetra Min, Tetra Werke, Melle

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

Control: 21.1 - 21.9 °C
Test groups: 21.1 - 21.9 °C

pH:

Control: 6.0 - 8.2
Test groups: 6.0 -

Dissolved oxygen:

Control: 6.9 - 9.2 mg/L
Test groups: 7.6 - 8.9 mg/L

Nominal and measured concentrations:

Nominal mg/L: 0; 10; 100; 500

Details on test conditions:

Test Facility
The experimental procedure was carried out in a static system. Calibrated test containers (10 liters) were made of glass (length 30 cm, width 22 cm, height 24 cm) and were immersed in a water bath with a Hostalit Z viewing window made of Plexiglas. The water bath was controlled via a thermostat at 22 ± 1°C. The basins were maintained in a day-night rhythm of 12 hours from the top. The illumination (700 lux) was directly above the basins during the entire experimental time, with no aeration.

Experimental procedure.
Following preparation of the test concentrations and subsequent measurement of water parameters, 10 fish were randomly distributed into the test and control tanks. During the entire experimental period the fish were not fed. After 2-4, 24, 48, 72 and 96 hours, the fish were checked for lethality and any visible changes in appearance and behavior were noted. Dead fish were removed from the tank. The fish were considered dead if no breathing movements were recognized, and also if the fish were unresponsive to mechanical stimulus. At the same time, the water parameters were measured and also logged.

Reference substance (positive control):

not required

Remarks:

for fish studies

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

pH 7.9 - 8.1
2-4 and 24 hours :after start of the experiment occurred in 10 / 100 and 500 mg / l - increased respiration. At 500 mg / l it was observed at the end of the experiment that some fish were swimming at the water surface. The fish of 10 and 100 mg / l group were symptom free after 48 hours from start of the experiment.

pH 6.0
500 mg / l: showed the following symptoms: tail-swimming position, prominence of the gill cover, increased breathing and lateral position on the pelvic floor. The dead fish were discolored.
100 mg / l: no behavioural effects.

Results with reference substance (positive control):

not required

Validity criteria fulfilled:

yes

Conclusions:

The strong points of this study are that 1) the study was conducted following OECD guidelines, 2) under GLP, and 3) that the substance was tested up to concentrations higher than its solubility. The weak points mainly relate to the lack of provision of information on the purity and physical-chemical properties of the compound, the lack of maintenance of the pH at 6 (in the studies where the pH was set at 6). At the limit concentration of 100 mg/L, there was no lethality observed, thus the LC50 is > 100 mg/L.

Executive summary:

96-Hour Acute Toxicity Study in Zebrafish (Brachydanio renio) with TANFLIX AL.

 

The study procedures described in this report were based on the EEC directive 84/449, Part C.1.'Acute toxicity for fish’; and the OECD guideline No. 203: "Fish Acute Toxicity Test’’, Adopted 04 April 1984.

 

The test was conducted at pH 7.9 – 8.1 (concentrations 10, 100 and 500 mg/L) and at pH 6 (100 and 500 mg/L). At pH 6, the solubility of Tanflix AL was better than at pH 7.9-8.1.  The experiment was performed in 4 parts, with 10 fish per test group. The following concentrations were tested:
Test 1:  0, 500 mg/L                 Begin date: 04.08.86

Test 2:  0, 500 mg/L                 Begin date: 18.08.86

Test 3:  0, 0, 10, 100 mg/L      Begin date: 25.08.86

Test 4: 0, 100 mg/L                  Begin date: 08.09.86

 

Only at pH 6 and at 500 mg/L were all the 10 fish dead, with no mortality recorded at 1, 10, and 100 mg/L. At pH 6, and at pH 7.8-9.1 the LC50 is greater than 100 mg/L. 

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07-06-2004 to 12-08-2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study is complete

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Sampling from the test vessels was performed at middle depth of the water column at
test start and after 24 h, 48 h, 72 h and 96 h. At 24 h, 48 h, and 72 h, the medium was
exchanged and both, new and old media were sampled. Samples were taken from
the test vessels using a plastic syringe and transferred into disposable 20 mL-polyethylene
vials. One subsample of 20 mL was acidified with 0.2 mL 14 M nitric acid,
thoroughly mixed and analysed without further treatment. The added amount of nitric
acid was sufficient to reduce the pH below 2. This fraction is designated as “unfiltered
sample” and is assumed to contain the “total aluminium”. A second subsample was
passed through a Sartorius 0.45 μm-membrane filter (type Minisart NML, diameter
26 mm). In order to rinse the filter, approx. 2 - 3 mL of the sample water was processed
at first. The filtrate from this rinsing was discarded. Then, further 20 mL of the
sample water was processed. This filtrate was acidified as described above (20 mL
sample plus 0.2 mL 14 M nitric acid; resulting pH below 2). In aquatic chemistry, 0.45
μm-membrane filtered samples generally are assumed to contain the “dissolved
metal” fractions.

Vehicle:

no

Details on test solutions:

Before test start and at each renewal, adequate amounts of the test item were given
in the new test vessels, filled up to the end volume with reconstituted water and continuously
stirred by magnetic stirrers. After one hour, the pH was adjusted to 7.5 in
each vessel by using NaOH. The fish were introduced and a sample for chemical
analysis was taken directly thereafter. The nominal test concentrations were 62.5,
125, 250, 500, and 1000 mg/L. Reconstituted water was tested as control.
The solutions were slightly aerated via a glass capillary. 10 fish were used for each
test concentration and for the control. The light/dark cycle was adjusted to 14/10
hours. The control group was held under the same conditions. No feeding occurred
during the test as well as 24 h before test start.
PH-value, oxygen concentration, and temperature were measured in the new vessels
directly before adding the fish and in the old vessels after transfer.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Danio rerio (Hamilton-Buchanan 1822) (Teleostei, Cyprinidae)
Length: 2 cm + 1 cm
Source: Laboratory bred
The strain of zebra fish was obtained from:
West Aquarium GmbH
PB 146
D-37431 Bad Lauterberg
The test fish were taken from cultures of the testing facility.
Holding: Test fish had been reared under study conditions (light, temperature)
in purified drinking water. They had been fed daily with Tetra-
Min® Hauptfutter (Tetra Werke, Melle, Germany).
Mortality: The criteria of the test guideline (ref 1) were followed. Only healthy
fish without diseases and abnormalities were used in the study.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Calcium chloride solution
Dissolve 11.76 g CaCl2*2H2O in deionised water;
make up to 1 litre with deionised water
Magnesium sulphate solution
Dissolve 4.93 g MgSO4*7H2O in deionised water;
make up to 1 litre with deionised water
Sodium bicarbonate solution
Dissolve 2.59 g NaHCO3 in deionised water; make up to 1 litre with deionised water
Potassium chloride solution
Dissolve 0.23 g KCl in deionised water; make up to 1 litre with deionised water
All chemicals must be of analytical grade.
The conductivity of the distilled or deionised water should not exceed 10 μScm-1.
25 mL each of solutions (a) to (d) are mixed and the total volume made up to 1 litre
with deionised water. The sum of the calcium and magnesium ions in this solution is
2.5 mmol/L. The proportion Ca:Mg ions is 4:1 and Na:K ions 10:1. The acid capacity
KS4.3 of this solution is 0.8 mmol/L.

Test temperature:

21.7-23.0 C

pH:

7.5-8.2

Dissolved oxygen:

86-96 % of saturation

Nominal and measured concentrations:

nominal test substance: 62.5, 125, 250, 500, 1000 mg/L
nominal Al: 5.67, 11.3, 22.7, 45.4, 90.1 mg/L
measured total Al (mean): 5.25, 10.6, 20.8, 43.2, 87.5 mg/L
measured dissolved Al (mean): 0.082, 0.104, 0.106, 0.127, 0.156 mg/L

Details on test conditions:

Prior to exposure all fish were measured for lengths and weights (Table 1). Fish
lengths were 2.2 to 2.7 cm and thus were within the range of sizes recommended by
the OECD guideline (ref 1) of 2 ± 1 cm. The mean weight resulted in a loading of
0.22 g/L test liquid.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks:

and other signs of intoxication

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.156 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Remarks:

Aluminium

Basis for effect:

mortality (fish)

Remarks:

and other signs of intoxiation

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 0.156 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Remarks:

Alumnium

Basis for effect:

mortality (fish)

Details on results:

Since in the controls a measurable unstructured spectroscopic background was detected
by ICP-OES measurements, all data were corrected by subtraction of this
background. The background corresponded to approx 0.2 mg/L Al and resulted
probably from the test medium (re-constituted water prepared by addition of salts to
de-ionised water, adjustment of the pH value after test item addition). The background
corresponded to a maximum value of 4 % of the lowest concentration analysed
in the samples. Blank solutions prepared from purified water showed no such
background For all concentrations all measured total aluminium concentrations ranged from 86 %
to 104 % of the nominal values.
The 0.45 μm-filtered fraction, defined as dissolved aluminium, was significantly
lower than the total aluminium fraction. The range of dissolved Alconcentrations
was < 0.1 – 0.4 mg/L Al for all Al-concentration levels. The highest
concentration seems to correspond to the Al-dissolution capacity of the test medium
(under the conditions applied, i.e. that the test media was buffered after addition of
the test item to prevent a change of the pH). At any time the dissolved Al concentration
was below the EC10 of 0.58 mg dissolved Al/L calculated for Gilufloc 40 H (Polyaluminumchloride)
exposure under static conditions without pH adjustment (study CEF-002/4-11)

Reported statistics and error estimates:

The analyses were performed in order to determine the total and the dissolved aluminium
concentration in water column samples. From the Al concentration the respective
Gilufloc 40 H (Polyaluminumchloride)-concentration was calculated.
The determination of the no effect concentration (NOEC) was based on nominal test
item concentrations, since the mean test item concentrations, calculated from the
measured total aluminium concentrations, deviated by < 20 % from the nominal concentrations.

Sublethal observations / clinical signs:

No signs of intoxication were observed

Validity criteria fulfilled:

yes

Conclusions:

The NOEC, based on the test substance, was determined to be >= 1000 mg/L nominal. Based on Expert judgement the NOEC for measured dissolved Al was >= 0.357 mg/L, The EC50 was > 0.357 mg/L. All criteria are met, thus the test is valid.

Executive summary:

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of Gilufloc 40 H (Polyaluminumchloride) to zebrafish (Danio rerio) according to OECD TG 203. Gilufloc 40 H (Polyaluminumchloride) was tested at nominal concentrations of 62.5, 125, 250, 500, and 1000 mg/L under semi-static conditions (daily renewal of the test medium) for 96 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted at 7.5 at each renewal. The water column was stirred during the test. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. Up to the highest test concentration, no mortality could be observed after 96 h. Thus, the NOEC of Gilufloc 40 H (Polyaluminumchloride) under the described study conditions was determined as follows: NOEC ≥ 1000 mg Gilufloc 40 H / L. The NOEC for dissolved Al was added by the reviewer to be >= 0.357 mg/L, the EC50 was > 0.357 mg/L.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Study period:

06-05-2003 to 08-08-2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: complete

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Sampling from the test vessels was performed at middle depth of the water column at
test start and after 24 h, 48 h, 72 h and 96 h. At 24 h, 48 h, and 72 h, the medium
was exchanged and both, new and old media were sampled. Samples were taken
from the test vessels using a plastic syringe and transferred into disposable 20 mLpolyethylene
vials. One subsample of 20 mL was acidified with 0.2 mL 14 M nitric
acid, thoroughly mixed and analyzed without further treatment. The added amount of
nitric acid was sufficient to reduce the pH below 2. This fraction is designated as “unfiltered
sample” and is assumed to contain the “total aluminium”. A second subsample
was passed through a Sartorius 0.45 μm-membrane filter (type Minisart NML, diameter
26 mm). In order to rinse the filter, approx. 2 - 3 mL of the sample water was
processed at first. The filtrate from this rinsing was discarded. Then, further 20 mL of
the sample water was processed. This filtrate was acidified as described above (20
mL sample plus 0.2 mL 14 M nitric acid; resulting pH below 2). In aquatic chemistry,
0.45 μm-membrane filtered samples generally are assumed to contain the “dissolved
metal” fractions.

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Danio rerio (Hamilton-Buchanan 1822) (Teleostei, Cyprinidae)
Length: 2 cm + 1 cm
Source: Laboratory bred
The strain of zebra fish was obtained from:
West Aquarium GmbH
PB 146
D-37431 Bad Lauterberg
The test fish were taken from cultures of the testing facility.
Holding: Test fish had been reared under study conditions (light, temperature)
in purified drinking water. They had been fed daily with Tetra-
Min® Hauptfutter (Tetra Werke, Melle, Germany).
Mortality: The criteria of the test guideline (ref 1) were followed. Only healthy
fish without diseases and abnormalities were used in the study.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

22.3 +- 0.3 C

pH:

7.4-8.0

Dissolved oxygen:

86-94% of saturation

Nominal and measured concentrations:

nominal: 62.5, 125, 250, 500, 1000 mg/L
measured (Al total):4.87, 10.4, 21.6, 43.4, 85.9 mg/L
measured (Al dissolved): 0.247, 0.205, 0.166, 0.131, 0.105 mg/L

Details on test conditions:

The test fish were exposed to the test item under semi-static conditions for a period
of 96 hours. Each day, the fish were transferred to freshly prepared test liquids in
new test vessels. The temperature during the test was 22.3 ± 0.3 °C. Mortality and
abnormal behaviour were recorded after 24 h, 48 h, 72 h and 96 h. 10 L glass
aquaria were used as test vessels. The loading of fish was 0.2 g/L and met the criteria
of the OECD-guideline.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Results indicate that the toxicity endpoint exceeds the water solubility of the test substance at the tested pH

Validity criteria fulfilled:

yes

Conclusions:

The NOEC for Aluminium sulphate -14-hydrate on Zebrafish >= 1000 mg/L nominal. It exceed the water solubility at pH 7.4-8 used in the test

Executive summary:

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of aluminium sulphate-14-hydrate to zebrafish (Danio rerio) according to OECD TG 203. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 62.5, 125, 250, 500, and 1000 mg/L under semi-static conditions (daily renewal of the test medium) for 96 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted at 7.5 at each renewal. The water column was stirred during the test. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. Up to the highest test concentration, no mortality could be observed after 96 h. Thus, the NOEC of aluminium sulphate-14-hydrate under the described study conditions was determined as follows: NOEC ≥ 1000 mg aluminium sulphate-14-hydrate / L, and > the water solubility at the tested pH range

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Study period:

06-05-2003 to 08-08-2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: methodology is clear and complete

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Sampling from the test vessels was performed at middle depth of the water column at
test start and after 96 h. Samples were taken from the test vessels using a plastic syringe
and transferred into disposable 20 mL-polyethylene vials. One subsample of 20
mL was acidified with 0.2 mL 14 M nitric acid, thoroughly mixed and analyzed without
further treatment. The added amount of nitric acid was sufficient to reduce the pH below
2. This fraction is designated as “unfiltered sample” and is assumed to contain the
“total aluminium”. A second subsample was passed through a Sartorius 0.45 μmmembrane
filter (type Minisart NML, diameter 26 mm). In order to rinse the filter, approx.
2 - 3 mL of the sample water was processed at first. The filtrate from this rinsing
was discarded. Then, further 20 mL of the sample water was processed. This filtrate
was acidified as described above (20 mL sample plus 0.2 mL 14 M nitric acid; resulting
pH below 2). In aquatic chemistry, 0.45 μm-membrane filtered samples generally
are assumed to contain the “dissolved metal” fractions.

Vehicle:

no

Details on test solutions:

Adequate amounts of the test item were given in the test vessels, filled up to the end
volume with reconstituted water and mixed thoroughly. The fish were introduced and
a sample for chemical analysis was taken directly thereafter.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Danio rerio (Hamilton-Buchanan 1822) (Teleostei, Cyprinidae)
Length: 2 cm + 1 cm
Source: Laboratory bred
The strain of zebra fish was obtained from:
West Aquarium GmbH
PB 146
D-37431 Bad Lauterberg
The test fish were taken from cultures of the testing facility.
Holding: Test fish had been reared under study conditions (light, temperature)
in purified drinking water. They had been fed daily with Tetra-
Min® Hauptfutter (Tetra Werke, Melle, Germany).
Mortality: The criteria of the test guideline were followed. Only healthy
fish without diseases and abnormalities were used in the study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Calcium chloride solution
Dissolve 11.76 g CaCl2*2H2O in deionised water;
make up to 1 litre with deionised water
Magnesium sulphate solution
Dissolve 4.93 g MgSO4*7H2O in deionised water;
make up to 1 litre with deionised water
Sodium bicarbonate solution
Dissolve 2.59 g NaHCO3 in deionised water; make up to 1 litre with deionised water
Potassium chloride solution
Dissolve 0.23 g KCl in deionised water; make up to 1 litre with deionised water
All chemicals must be of analytical grade.
The conductivity of the distilled or deionised water should not exceed 10 μScm-1.
25 mL each of solutions (a) to (d) are mixed and the total volume made up to 1 litre
with deionised water. The sum of the calcium and magnesium ions in this solution is
2.5 mmol/L. The proportion Ca:Mg ions is 4:1 and Na:K ions 10:1. The acid capacity
KS4.3 of this solution is 0.8 mmol/L.

Test temperature:

21.9-22.7 C

pH:

ph was dependent of test substance concentration: Control; 7.8-8.2
Test concentrations 7.7-4.2 (increasing with increase with test substance concentration

Dissolved oxygen:

89-99% of saturation

Nominal and measured concentrations:

Nominal (testsubstance): 62.5, 80, 100, 125, 250, 500, 1000 mg/L
aluminium nominal: 5.63, 7.20, 9.00, 11.3, 22.5, 45.0, 90.0 mg/L
mean measured aluminium in test solution: <0.10, 0.104, 0.795, 2.71, 14.1, 35.4, 82.7 mg/L

Details on test conditions:

Since in the controls a measurable unstructured spectroscopic background was detected
by ICP-OES measurements, all data were corrected by subtraction of this
background. The background corresponded to approx 0.2 mg/L Al and resulted
probably from the test medium (re-constituted water prepared by addition of salts to
de-ionized water, no adjustment of pH after test item addition). The background corresponded
to a maximum value of 5 % of the lowest concentration analysed in the
samples. Blank solutions prepared from purified water showed no such background.
Furthermore, no background was detected in the membrane-filtered control samples
analysed by ICP-MS for determination of the dissolved aluminium fraction.
For all concentrations only slight deviations of the measured total aluminium concentrations
from the nominal values were observed. In all freshly prepared test media at study
start, concentrations were between 96 and 101 % of the nominal values. At study termination
for the respective vessel, concentrations were still between 79 and 98 % of the
nominal total aluminium concentrations, most probably due to the water movement by
aeration and swimming fish. The mean measured total aluminium concentrations in the
test media (90 to 98 % of nominal) deviated by less than 20 % from nominal values.
Thus, nominal concentrations were taken as the basis for the determination of effect
concentrations. Differences between study start and end were only low,
especially for the concentration levels above 100 mg test item/L.
The reason for the high amounts of aluminium dissolved is that the pH-values in the
test media were not controlled. Since the dissolution of the test item aluminium sulphate-
14-hydrate lowers the pH of the solution, the solubility of Al is increased.
This effect is highest at the highest concentration level tested.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

98 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

104 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 93-117 mg/L

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

8.8 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

other: total aluminium

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

9.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

other: total aluminium

Basis for effect:

mortality (fish)

Remarks on result:

other: 8.4-11

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

0.71 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

dissolved

Remarks:

aluminium

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

dissolved

Remarks:

aluminium

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.45-2.2 mg/L

Reported statistics and error estimates:

The evaluation of the effect concentrations was based on nominal test item concentrations,
since the mean test item concentrations, calculated from the mean measured
total aluminium concentrations, deviated by < 20 % from the nominal concentrations.
The evaluation of the concentration-effect-relationships and the calculations of effect
concentrations of dissolved aluminium were based on the mean measured concentrations.
LC10 and LC50 values were determined together with 95% confidence intervals using
Probit-analysis assuming log-normal distribution of the values by using the
ToxRat programme.

Sublethal observations / clinical signs:

cumulative mortaility during the test:

Test part	Nominalconcentration mg/L	Test duration
		6h	24h	48h	72h	96h
A	Control	0	0	0	0	0
B	Control	0	0	0	0	0
A	62.5	0	0	0	0	0
B	80	0	0	0	0	0
B	100	0	0	2	2	2
A	125	0	2	9	10	10
A	250	2	10	10	10	10
A	500	1	10	10	10	10
A	1000	2	10	10	10	10
B	pH 5.0	0	0	0	0	0
B	pH 4.8	0	0	0	0	0
B	pH 4.5	0	0	0	0	0
B	pH 4.3	0	0	0	0	0

 

 

Validity criteria fulfilled:

yes

Conclusions:

Thus, the LC50 of the test item was determined to be 104 mg test item /L (95 % confidence limits: 93 – 117 mg/L). 104 mg/L of aluminium sulphate-14-hydrate reduced the pH of reconstituted water (OECD 203) to a level which enabled 1.0 mg Al/L to be dissolved, causing 50 %
mortality after 96 h.

Executive summary:

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of aluminium sulphate-14-hydrate to zebrafish (Danio rerio) relevant for an accidental exposure. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 62.5, 80, 100, 125, 250, 500, and 1000 mg/L under static conditions for 96 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. To clarify whether mortalities at concentrations of 100 to 500 mg/L are due to the low pH or the influence of aluminium, the initial pH at the respective concentrations was adjusted to additional test vessels by using sulfuric acid but without adding aluminium sulphate. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. The mean measured dissolved aluminium concentrations increased with increasing test item concentration from below 2 % to 94 % of the total Al-concentration, caused by the decrease of pH by the test item, increasing the solubility of aluminium. The LC50 of the test item was determined to be - 104 mg test item /L (95 % confidence limits: 93 – 117 mg/L) The very steep concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration: 104 mg/L of aluminium sulphate-14-hydrate reduced the pH of reconstituted water (OECD 203) to a level which enabled 1.0 mg Al/L to be dissolved, causing 50 % mortality after 96 h.

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1998

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Test according to OECD guideline with very limited reporting on methodology. Klimisch rating may improve if complete report becomes available.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

Principles of method if other than guideline:

Not relevant

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

not specified

Test type:

not specified

Water media type:

not specified

Limit test:

no

Dose descriptor:

NOEC

Effect conc.:

429.6 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Dose descriptor:

LC50

Effect conc.:

644.4 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Dose descriptor:

LC100

Effect conc.:

886.1 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

No data

Validity criteria fulfilled:

not specified

Conclusions:

The acute toxicity of AL2(SO4)3 towards fish was investigated according to OECD Guideline 203. The NOEC, LC50 and LC100 were found to be 429.6, 644.4 and 886.1 mg/l respectively.

Executive summary:

The acute toxicity of AL2(SO4)3 towards fish was investigated according to OECD Guideline 203. The NOEC, LC50 and LC100 were found to be 429.6, 644.4 and 886.1 mg/l respectively.

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07-06-2004 to 12-08-2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study is complete, one remark is the variance in pH due to the testsubstance

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Sampling from the test vessels was performed at middle depth of the water column at
test start and after 96 h. Samples were taken from the test vessels using a plastic syringe
and transferred into disposable 20 mL-polyethylene vials. One subsample of 20
mL was acidified with 0.2 mL 14 M nitric acid, thoroughly mixed and analysed without
further treatment. The added amount of nitric acid was sufficient to reduce the pH below
2. This fraction is designated as “unfiltered sample” and is assumed to contain the
“total aluminium”. A second subsample was passed through a Sartorius 0.45 μmmembrane
filter (type Minisart NML, diameter 26 mm). In order to rinse the filter,
approx. 2 - 3 mL of the sample water was processed at first. The filtrate from this rinsing
was discarded. Then, further 20 mL of the sample water was processed. This filtrate
was acidified as described above (20 mL sample plus 0.2 mL 14 M nitric acid;
resulting pH below 2). In aquatic chemistry, 0.45 μm-membrane filtered samples generally
are assumed to contain the “dissolved metal” fractions.

Nitric acid was of ‘Suprapur’ quality (supplied by VWR, Darmstadt). The purified water
used was processed using an ELGA UHQ/PS water purification system (water quality:
reciprocal conductivity > 18 Mohm*cm).
As aluminium standard a commercially available solution was used (1000 μg/mL Al in
2 % nitric acid, from VWR, Darmstadt, traceable to a NIST standard reference material).
From this standard solution appropriate stock solutions were prepared which
were used for the preparation of calibration solutions. Dilutions were performed with
0.5 M nitric acid in water.

Vehicle:

no

Details on test solutions:

Adequate amounts of the test item were given in the test vessels, filled up to the end
volume with reconstituted water and mixed thoroughly. The fish were introduced and
a sample for chemical analysis was taken directly thereafter.
The nominal test concentrations were 62.5, 125, 250, 500, and 1000 mg/L. Reconstituted
water was tested as control. The test liquids were slightly aerated via a glass
capillary. 10 fish were introduced in each test vessel. The light/dark cycle was adjusted
to 14/10 hours. The control group was held under the same conditions. No
feeding occurred during the test as well as 24 h before test start.
pH-value, oxygen concentration, and temperature were measured directly before adding
the fish and afterwards once per day.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Danio rerio (Hamilton-Buchanan 1822) (Teleostei, Cyprinidae)
Length: 2 cm + 1 cm
Source: Laboratory bred
The strain of zebra fish was obtained from:
West Aquarium GmbH
PB 146
D-37431 Bad Lauterberg
The test fish were taken from cultures of the testing facility.
Holding: Test fish had been reared under study conditions (light, temperature)
in purified drinking water. They had been fed daily with Tetra-
Min® Hauptfutter (Tetra Werke, Melle, Germany).
Mortality: The criteria of the test guideline (ref 1) were followed. Only healthy
fish without diseases and abnormalities were used in the study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Calcium chloride solution
Dissolve 11.76 g CaCl2*2H2O in deionised water;
make up to 1 litre with deionised water
Magnesium sulphate solution
Dissolve 4.93 g MgSO4*7H2O in deionised water;
make up to 1 litre with deionised water
Sodium bicarbonate solution
Dissolve 2.59 g NaHCO3 in deionised water; make up to 1 litre with deionised water
Potassium chloride solution
Dissolve 0.23 g KCl in deionised water; make up to 1 litre with deionised water
All chemicals must be of analytical grade.
The conductivity of the distilled or deionised water should not exceed 10 μScm-1.
25 mL each of solutions (a) to (d) are mixed and the total volume made up to 1 litre
with deionised water. The sum of the calcium and magnesium ions in this solution is
2.5 mmol/L. The proportion Ca:Mg ions

Test temperature:

21.7-22.8 C

pH:

4.2-8.3

Dissolved oxygen:

89-94% of saturation

Nominal and measured concentrations:

nominal test substance: 62.5, 125, 250, 500, 1000 mg/L
nominal Al: 5.67, 11.3, 22.7, 45.4, 90.7 mg/L
measured total Al: 5.37, 10.87, 22.2, 42.7, 87.2 mg/L
measured Al (aq): <0.1, <0.1, 8.58, 36.9, 90.8 mg/L

Details on test conditions:

Prior to exposure all fish were measured for lengths and weights. Fish
lengths were 2.0 to 2.7 cm and thus were within the range of sizes recommended by
the OECD guideline of 2 ± 1 cm. The mean weight resulted in a loading of
0.21 g/L test liquid.
Adequate amounts of the test item were given in the test vessels, filled up to the end
volume with reconstituted water and mixed thoroughly. The fish were introduced and
a sample for chemical analysis was taken directly thereafter.
The nominal test concentrations were 62.5, 125, 250, 500, and 1000 mg/L. Reconstituted
water was tested as control. The test liquids were slightly aerated via a glass
capillary. 10 fish were introduced in each test vessel. The light/dark cycle was adjusted
to 14/10 hours. The control group was held under the same conditions. No
feeding occurred during the test as well as 24 h before test start.
pH-value, oxygen concentration, and temperature were measured directly before adding
the fish and afterwards once per day.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

142 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

dissolved

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.41-4.78 mg/L

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

0.58 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

dissolved

Remarks:

Al

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

186 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 161-216 mg/L

Details on results:

Since in the controls a measurable unstructured spectroscopic background was detected
by ICP-OES measurements, all data were corrected by subtraction of this
background. The background corresponded to approx 0.2 mg/L Al and resulted
probably from the test medium (re-constituted water prepared by addition of salts to
de-ionised water, no adjustment of pH after test item addition). The background corresponded
to a maximum value of 4 % of the lowest concentration analysed in the
samples. Blank solutions prepared from purified water showed no such background.
For all concentrations only slight deviations of the measured total aluminium concentrations
from the nominal values were observed. All measurements at test start as
well as at test termination for the respective vessel yielded concentrations between
94 and 100 % of the nominal total aluminium concentrations. The mean measured total
aluminium concentrations in the test media (94 to 98 % of nominal) deviated by
less than 20 % from nominal values. Thus, nominal concentrations were taken as the
basis for the determination of effect concentrations.
The measured concentrations of the 0.45 μm-filtered fraction, defined as dissolved
aluminium were differing by three orders of magnitude. Expressed as percent
of total (unfiltered) Al-concentrations, it increased with increasing test item concentration
(i.e., at the nominal concentration of 62.5 mg/L the dissolved Al fraction
amounted to an average of 3 % of the total Al-concentration while for the highest
nominal concentration the dissolved fraction was equal to the total Al-concentration).
Differences between test start and end followed different trends. Whereas at the lowest
dose level the measured dissolved Al-concentration increased, concentrations
declined or remained constant at the higher dose levels.

Sublethal observations / clinical signs:

The reason for the high amounts of aluminium dissolved is that the pH-values in the test media were not controlled. Since the dissolution of the test item Gilufloc 40 H (Polyaluminiumchloride) lowers the pH of the solution, the solubility of Al is increased This effect is highest at the highest concentration level tested.

The cumulative mortality of fish during the test period of 96 h indicates a steep concentration-response relationship. Whereas the mean measured concentrations of dissolved aluminium causing 0 % and 100 % mortality differed by a factor of approximately 35, the respective total aluminium or test item concentrations differed only by a factor of 2. The much steeper concentration-response relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration

Table1: cumulative mortality

Nominal test substance concentration (mg/L)	Dissloved aluminium (mg/l)	Test duration
		5h	24h	48h	72h	96h
Control	(0.197)	0	0	0	0	0
62.5	<0.1	0	0	0	0	0
125	<0.1	0	0	0	0	0
250	6.04	0	10	10	10	10
500	26.2	4	10	10	10	10
1000	89.9	1	10	10	10	10

Validity criteria fulfilled:

yes

Conclusions:

The LC10 and LC50 were determined for the test compound to be 186 mg/L and 142 mg/L nominal, respectively. For dissolved aluminium the LC10 and LC50 were determined to be 1.39 and 0.58 mg/L, respectively. The test criteria were fulfilled and thus the study is valid. The very steep concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration

Executive summary:

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of Gilufloc 40 H (Polyaluminiumchloride) to zebrafish (Danio rerio) relevant for an accidental exposure. Gilufloc 40 H (Polyaluminiumchloride) was tested at nominal concentrations of 62.5, 125, 250, 500, and 1000 mg/L under static conditions for 96 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. The mean measured dissolved aluminium concentrations increased with increasing test item concentration from 2 % to 99 % of the total Al concentration. This was caused by the decrease of pH by the test item which increased the solubility of aluminium. The LC50 of the test item was determined to be 186 mg test item /L (95 % confidence limits: 161 – 216 mg/L). The very steep concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration: Theoretically, 186 mg/L of Gilufloc 40 H (Polyaluminiumchloride) reduced the pH of reconstituted water (OECD 203) to a level which enabled 1.4 mg Al/L to be dissolved, causing 50 % mortality after 96 h, already manifest after 24 h. The LC10 for the test compound was 142 mg/L nominal and for dissolved aluminium 0.58 mg/L.

Description of key information

OECD guideline 203, GLP, key study, validity 1 (semi-static test, Schafers, 2004):

96 -hour LC50 > 0.357 mg/L (based on nominal concentrations)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.357 mg/L

Additional information

Six read-across studies are available to evaluate the short-term toxicity to fish of the registered substance :

 

1) Key study on aluminium sulphate-14-hydrate, by Schafers (static test, 2003) :

 A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institut= for Molecular Biology and Applied Ecology to evaluate the acute toxicity of aluminium sulphate-14-hydrate to zebrafish (Danio rerio) relevant for an accidental exposure. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 62.5, 80, 100, 125, 250, 500, and 1000 mg/L under static conditions for 96 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. To clarify whether mortalities at concentrations of 100 to 500 mg/L are due to the low pH or the influence of aluminium, the initial pH at the respective concentrations was adjusted to additional test vessels by using sulfuric acid but without adding aluminium sulphate. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. The mean measured dissolved aluminium concentrations increased with increasing test item concentration from below 2 % to 94 % of the total Al-concentration, caused by the decrease of pH by the test item, increasing the solubility of aluminium. The LC50 of the test item was determined to be - 104 mg test item /L (95 % confidence limits: 93 – 117 mg/L) The very steep concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration: 104 mg/L of aluminium sulphate-14-hydrate reduced the pH of reconstituted water (OECD 203) to a level which enabled 1.0 mg Al/L to be dissolved, causing 50 % mortality after 96 h.

 

2) Key study on aluminium sulphate-14-hydrate, by Schafers (semi-static test, 2003)

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of aluminium sulphate-14-hydrate to zebrafish (Danio rerio) according to OECD TG 203. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 62.5, 125, 250, 500, and 1000 mg/L under semi-static conditions (daily renewal of the test medium) for 96 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted at 7.5 at each renewal. The water column was stirred during the test. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. Up to the highest test concentration, no mortality could be observed after 96 h. Thus, the NOEC of aluminium sulphate-14-hydrate under the described study conditions was determined as follows: NOEC ≥ 1000 mg aluminium sulphate-14-hydrate / L, and > the water solubility at the tested pH range.

 

3) Key study on aluminium trichloride 2004 (STATIC)

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of Gilufloc 40 H (Polyaluminiumchloride) to zebrafish (Danio rerio) relevant for an accidental exposure. Gilufloc 40 H (Polyaluminiumchloride) was tested at nominal concentrations of 62.5, 125, 250, 500, and 1000 mg/L under static conditions for 96 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. The mean measured dissolved aluminium concentrations increased with increasing test item concentration from 2 % to 99 % of the total Al concentration. This was caused by the decrease of pH by the test item which increased the solubility of aluminium. The LC50 of the test item was determined to be 186 mg test item /L (95 % confidence limits: 161 – 216 mg/L). The very steep concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration: Theoretically, 186 mg/L of Gilufloc 40 H (Polyaluminiumchloride) reduced the pH of reconstituted water (OECD 203) to a level which enabled 1.4 mg Al/L to be dissolved, causing 50 % mortality after 96 h, already manifest after 24 h. The LC10 for the test compound was 142 mg/L nominal and for dissolved aluminium 0.58 mg/L.

 

4) Key study on aluminium trichloride, by Schafers (semi-static test, 2004)

A study sponsored by the Aluminium Salts Subgroup of the Cefic Incopa Sector Group was performed at the Fraunhofer-Institute for Molecular Biology and Applied Ecology to evaluate the acute toxicity of Gilufloc 40 H (Polyaluminumchloride) to zebrafish (Danio rerio) according to OECD TG 203. Gilufloc 40 H (Polyaluminumchloride) was tested at nominal concentrations of 62.5, 125, 250, 500, and 1000 mg/L under semi-static conditions (daily renewal of the test medium) for 96 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted at 7.5 at each renewal. The water column was stirred during the test. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values. Up to the highest test concentration, no mortality could be observed after 96 h. Thus, the NOEC of Gilufloc 40 H (Polyaluminumchloride) under the described study conditions was determined as follows: NOEC ≥ 1000 mg Gilufloc 40 H / L. The NOEC for dissolved Al was added by the reviewer to be >= 0.357 mg/L, the EC50 was > 0.357 mg/L.

 

5) Study on aluminium sulphate, by Benrens (1998)

 The acute toxicity of AL2(SO4)3 towards fish was investigated according to OECD Guideline 203. The NOEC, LC50 and LC100 were found to be 429.6, 644.4 and 886.1 mg/l respectively.

 

6) Study on Aluminium hydroxy chloride, by Markert (1986)

96-Hour Acute Toxicity Study in Zebrafish (Brachydanio renio) with TANFLIX AL.

The study procedures described in this report were based on the EEC directive 84/449, Part C.1.'Acute toxicity for fish’; and the OECD guideline No. 203: "Fish Acute Toxicity Test’’, Adopted 04 April 1984.

The test was conducted at pH 7.9 – 8.1 (concentrations 10, 100 and 500 mg/L) and at pH 6 (100 and 500 mg/L). At pH 6, the solubility of Tanflix AL was better than at pH 7.9-8.1.  The experiment was performed in 4 parts, with 10 fish per test group. The following concentrations were tested: Test 1:  0, 500 mg/L                 Begin date: 04.08.86

Test 2:  0, 500 mg/L                 Begin date: 18.08.86

Test 3:  0, 0, 10, 100 mg/L      Begin date: 25.08.86

Test 4: 0, 100 mg/L                  Begin date: 08.09.86

Only at pH 6 and at 500 mg/L were all the 10 fish dead, with no mortality recorded at 1, 10, and 100 mg/L. At pH 6, and at pH 7.8-9.1 the LC50 is greater than 100 mg/L.