Reaction mass of aluminium hydroxide and aluminium nitrate and aluminium sulphate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles.

Qualifier:

no guideline followed

Principles of method if other than guideline:

Juvenile Atlantic salmon (Salmo salar) were exposed to Al in either inorganic or organic acidic soft waters, at pH 5.0, for 96 and 120 h and LC50 values determined.

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

Samples were taken daily. Acid-washed plasticware was used.

Vehicle:

no

Details on test solutions:

A commercially available soil FA (Laurentian FA: Ecolinc Inc., Roxboro, QC, Canada) was chosen to represent the natural DOM found in rivers on the Canadian Shield during periods of snowmelt. This FA is obtained from soil collected from the Laurentian region north of the St. Lawrence River.
A 20 mg FA/mL stock solution was prepared by dissolving a weighed quantity of FA in dionized water. The experimental FA solutions were prepared in 65 L basins fitted with plastic liners. Reconstituted soft water was used (Ca2+: 44uM; Mg2+: 62 uM; Na+: 143uM; K+: 7 uM; Cl-: 7 uM; [SO4]2-: 106 uM; [CO3]2-: 143 uM. pH was adjusted to ≤ 4.5 with 2.0 N HNO3 and gentle aeration was provided. Appropriate amounts of the FA stock solution and Al stock solution (1000 mg/L Al as Al2[SO4]3 x 18H2O, acidified to

Test organisms (species):

Salmo salar

Details on test organisms:

TEST ORGANISM
- Common name: Atlantic salmon
- Source: provincial fish hatchery (Ministére de Loisir, Chasse et Pêche, Gaspé, QC, Canada) and from a commercial aquaculture establishment (La Pisciculture Saukeb, Nouvele Gaspé, QC, Canada). Stocks of salmon originated from rivers of the Gaspé, (MLCP) and North Shore (La Pisciculture Saukeb) regions of Québec.
- Weight at study initiation: average wet weight 1.8 ± 0.8 g, range 0.9 - 3.1 g
- Feeding during test: no; not fed for 24 h prior to an experiment


ACCLIMATION
- Acclimation period: at least 2 weeks
- Acclimation conditions (same as test or not): no; fish were maintained in 550 L holding tanks, with daily renewal of water; temp. 10°C; dissolved oxygen >95% saturation; 12 h: 12 h light-dark cycle
- Type and amount of food: trout chow pellets (Corey Mills, Fredericton, NB, Canada)
- Feeding frequency: ad libitum

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

120

Remarks on exposure duration:

96 h in experiment 1; 120 h in experiment 2

Hardness:

n/a

Test temperature:

10 °C

pH:

nominal: 5
Al exposure, experiment 1: 4.96 ± 0.16
Al exposure, experiment 2: 4.98 ± 0.06
Al and fulvic acid exposure, experiment 1: 5.03 ± 0.13
Al and fulvic acid exposure, experiment 2: 4.97 ± 0.18

Dissolved oxygen:

> 85% saturation

Nominal and measured concentrations:

Al exposure:
experiment 1: 3.9 ± 0.1; 3.2 ± 0.1; 3.4 ± 0.1; 2.2 ± 0.1; 1.1 ± 0.1; control 0.4 ± 0.1 (µM total Al, average measured)
experiment 2: 7.8 ± 0.8; 3.9 ± 0.3; 3.6 ± 0.2; 2.4 ± 0.2; 1.0 ± 0.2; control 0.1 ± 0.1 (µM total Al, average measured)

Al and fulvic acid exposure:
experiment 1: 38.6 ± 3.3; 33.8 ± 3.7; 20.8 ± 0.7; 10.5 ± 0.9 (µM total Al, average measured); 44.4, 33.3, 22.2, 11.1 (nominal)
experiment 2: 37.1 ± 10.1; 28.0 ± 3.8; 23.4 ± 2.8; 10.3 ± 2.3 (µM total Al, average measured); 44.4, 33.3, 22.2, 11.1 (nominal)

experiment 3: preexposure followed by Al exposure
five concentrations: 2.0 - 8.0 µM

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 7 or 8 L
- Renewal rate of test solution (frequency/flow rate): > 99% molecular replacement of the basin volume within 24 h
- No. of organisms per vessel: 7 (experiment 1), 8 (experiment 2)
- No. of vessels per concentration (replicates): 4-5
- No. of vessels per control (replicates): 4-5

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: RSW was prepared by the addition of salts to deionized water
- Total organic carbon: 1-3 mg C/L

OTHER TEST CONDITIONS
- Photoperiod: 12 hr light : 12 hr dark

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

78.3 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total inorganic Al (inorganic medium)

Basis for effect:

mortality (fish)

Remarks on result:

other: experiment 1; pH 4.96 ± 0.16; 95% CL: 67.5 - 86.4; LC50 re-calculated to µg/L by submitter based on data given in original reference

Duration:

96

Dose descriptor:

LC50

Effect conc.:

224.1 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total inorganic Al (fulvic acid solution)

Basis for effect:

mortality (fish)

Remarks on result:

other: experiment 1; pH 4.96 ± 0.16; 95% CL: 186.3 - 270; LC50 re-calculated to µg/L by submitter based on data given in original reference

Duration:

120 h

Dose descriptor:

LC50

Effect conc.:

108 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total inorganic Al (inorganic medium)

Basis for effect:

mortality (fish)

Remarks on result:

other: experiment 2; pH 4.98 ± 0.06; 95% CL: 94.5 - 121.5; LC50 re-calculated to µg/L by submitter based on data given in original reference

Duration:

120 h

Dose descriptor:

LC50

Effect conc.:

164.7 µg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total inorganic Al (folvic acid solution)

Basis for effect:

mortality (fish)

Remarks on result:

other: experiment 2; pH 4.98 ± 0.06; 95% CL: 156.6 - 221.4; LC50 re-calculated to µg/L by submitter based on data given in original reference

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

06-05-2003 to 08-08-2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to an appropriate OECD test guideline, and in compliance with GLP. The study is considered to be reliability 1 (reliable without restrictions); the read across of the result is considered to be reliability 2 (reliable with restrictions).

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 3

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

06-05-2003 to 08-08-2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to an appropriate OECD test guideline, and in compliance with GLP. The study is considered to be reliability 1 (reliable without restrictions); the read across of the result is considered to be reliability 2 (reliable with restrictions).

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 4

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented study which meets basic scientific principles.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Determination of the effects of humic acid and hardness on acute aluminum toxicity in flow-through laboratory tests with juvenile rainbow trouts in the neutral alkaline pH range. Mortality and growth were chosen as endpoints of toxicity.

GLP compliance:

no

Details on sampling:

Aluminum samples and either hardness or humic acid determinations were taken every 48 h.

Vehicle:

no

Details on test solutions:

Well water was processed by reverse osmosis (RO) reducing total dissolved solids by about 95% (hardness = 8 - 12 mg/L as CaCO3). Prior to toxicity tests, storage tank water was reconstituted to the proper chemical composition.
An aluminium stock was maintained at 16 mg Al/L with a pH of 9.0. The composition of the dilution water was 80 mg/L NaHCO3, 4 mg/L KCl, 10 mg/L CaSO4, and 5 mg/L MgSO4.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: Oregon State University marine and Freshwater Biomedical Center Core Facility, USA
- Age at study initiation: juvenile
- Weight at study initiation: 1 - 3 g
- Method of housing: 300 L fiber-glass tanks supplied with running well water; fed Oregon Test Diet (Sinnhuber et al 1977, Neoplasms in rainbow trout, Ann. N.Y. Acad. Sci 298: 389-408) ad libitum until 24h prior to acute tests
- Feeding during test: no

ACCLIMATION
- Acclimation conditions: same general conditions as tests
- Type and amount of food: Oregon Test Diet (Sinnhuber et al. 1977) ad libitum
- Feeding frequency: fed until 24 hours prior to acute exposure

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

10, 30, 80 and 120 mg/L as CaCO3 (nominal) in different test series

Test temperature:

14 ± 0.5 °C

pH:

7.3 and 8.3 in different tests

Dissolved oxygen:

Measured but not reported

Nominal and measured concentrations:

0, 1, 2, 4, and 8 mg/L (nominal Al)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 20 L aquaria
- Type of flow-through: A circulation pump ensured proper mixing of solutions; continuous-flow Chadwick-type diluter
- Renewal rate of test solution (frequency/flow rate): 100 mL / min, resulting in a volume replacement time of 3.3 h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6.17 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 5.33-7.25; average hardness 23.2 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6.17 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 5.31-7.20; average hardness 35 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

7.67 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 6.22-9.61; average hardness 83.6 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6.93 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: not reliabl5; average hardness 115.8 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.43 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 4.69-6.27; average humic acid concentration 2.6 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 3.68-5.74; average humic acid concentration 6.6 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.22 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

element

Remarks:

Al

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 4.44-6.14; average humic acid concentration 10.1 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.57 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.51-0.64; average hardness 23.2 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.61 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.51-0.73; average hardness 35 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.73 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.65-0.94; average hardness 83.6 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.67 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: not reliable; average hardness 115.8 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.36 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.31-0.417; average humic acid concentration 1.4 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.63 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.56-0.70; average humic acid concentration 2.6 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.62 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.53-0.72; average humic acid concentration 6.6 mg/L; pH 8.3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.79 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: CL: 0.67-0.92; average humic acid concentration 10.1 mg/L; pH 8.3

Details on results:

- There was no mortality at any of the exposure concentrations at nominal pH 7.3

Reported statistics and error estimates:

Toxicity was analyzed using the trimmed Spearman-Karber method for 96-h LC50 determinations (Hamilton et al. 1977, Trimmed Spearman-Karber method for estaimating median lethal concentrations in toxicity bioassays, Environ. Sci. technol. 11: 714-719) and pairs of LC50s were considered different when confidence limits did not overlap and when the ratio between the greater and lesser LC50s exceeded the calculated f ratio (Litchfield and Wilcoxon 1949, A simplified method of evaluating dose-effect experiments, J. Pharmacol. Exp. Ther. 96: 99-113). Statgraphics (1986) and SAS Institute Inc. (1985) PC software was used for statistical analyses.

Sublethal observations / clinical signs:

Results:

Table 1: Summary of test conditions and percent survival in exposure tanks for 96h acute tests at weakly alkaline pH (pH 8.39) [mean ± SE from duplicate tests]. The mean pH and aluminum concentrations were not significantly different (p > 0.05) between exposure tanks maintained at different hardness and the same nominal aluminium concentration.

pH	Total hardness (mg CaCO3/L)	Aluminium (mg/L)			% survival
		Nominal	Total	Filterable
7.97±0.03	23.1±0.1	0	<0.01	<0.01	100
8.02±0.03	34.2±3.9	0	<0.01	<0.01	100
8.06±0.03	83.3±4.4	0	<0.01	<0.01	100
8.06±0.04	112.5±6.1	0	<0.01	<0.01	100
8.12±0.03	21.9±0.7	1	0.81±0.30	0.11±0.04	100
8.10±0.02	33.1±3.5	1	1.02±0.25	0.09±0.04	100
8.23±0.03	84.2±4.3	1	0.91±0.07	0.16±0.03	100
8.25±0.03	144.5±6.7	1	1.05±0.09	0.20±0.02	100
8.22±0.02	22.9±2.3	2	1.86±0.47	0.18±0.08	100
8.20±0.03	33.9±4.0	2	2.04±0.57	0.21±0.05	100
8.23±0.03	84.2±4.5	2	1.92±0.71	0.16±0.03	100
8.25±0.04	114.5±5.3	2	1.68±0.59	0.20±0.10	100
8.34±0.03	23.4±2.5	4	3.73±0.75	0.42±0.18	85
8.36±0.03	36.3±3.2	4	4.32±1.12	0.46±0.15	85
8.36±0.03	83.0±4.7	4	4.17±1.12	0.42±0.14	95

Table 2: Summary of test conditions and percent survival in exposure tanks for 96h acute tests at near-neutral pH (ph 8.39) [mean ± SE from duplicate tests]. The mean pH and aluminum concentrations were not significantly different (p > 0.05) between exposure tanks maintained at different hardness and the same nominal aluminum concentration. Survival was 100% in all cases.

pH	Total hardness (mg CaCO3/L)	Aluminium (mg/L)
		Nominal	Total	Filterable
7.60±0.04	25.6±2.4	0	< 0.01	< 0.01
7.55±0.03	46.4±2.1	0	< 0.01	< 0.01
7.58±0.03	86.6±4.7	0	< 0.01	< 0.01
7.60±0.03	124.8±7.7	0	< 0.01	< 0.01
7.62±0.04	25.3±2.9	1	0.74±0.16	0.14±0.05
7.60±0.03	45.8±1.5	1	0.78±0.11	0.09±0.01
7.60±0.04	86.8±4.6	1	0.72±0.06	0.10±0.01
7.60±0.03	126.0±6.1	1	1.08±0.20	0.10±0.01
7.64±0.03	25.6±2.5	2	2.12±0.33	0.14±0.05
7.62±0.03	45.6±1.8	2	2.06±0.30	0.12±0.02
7.62±0.03	87.5±4.0	2	1.68±0.06	0.12±0.02
7.64±0.03	127.6±6.4	2	1.73±0.12	0.11±0.01
7.63±0.04	25.6±2.8	4	4.30±0.32	0.15±0.04
7.62±0.03	44.6±1.0	4	4.06±0.53	0.13±0.02
7.61±0.03	86.9±2.6	4	4.71±0.51	0.11±0.01
7.60±0.03	128.0±5.3	4	4.64±0.70	0.15±0.02
7.58±0.03	27.4±2.1	8	9.84±0.47	0.17±0.05
7.62±0.03	45.2±2.7	8	8.07±0.92	0.15±0.03
7.58±0.04	89.5±4.8	8	8.16±0.79	0.14±0.02
7.58±0.04	130.4±6.9	8	8.20±1.14	0.16±0.04

Table 3: Summary of test conditions and percent survival in exposure tanks for 96h acute tests at weakly alkaline pH (ph 8.39) [mean ± SE from duplicate tests]. The mean pH and aluminum concentrations were not significantly different (p > 0.05) between exposure tanks maintained at different humic acids and the same nominal aluminum concentration.

pH	Total hardness (mg CaCO3/L)	Aluminium (mg/L)			% survival
		Nominal	Total	Filterable
±0.06	0.92±0.08	0	< 0.01	< 0.01	100
8.05±0.06	2.24±0.05	0	< 0.01	< 0.01	100
7.95±0.04	6.21±0.29	0	< 0.01	< 0.01	100
7.96±0.03	9.72±0.03	0	< 0.01	< 0.01	100
8.11±0.06	1.19±0.22	1	0.83±0.09	0.14±0.02	100
8.11±0.06	2.38±0.15	1	0.86±0.05	0.17±0.03	100
8.03±0.05	6.48±0.24	1	0.86±0.06	0.20±0.04	100
7.98±0.04	10.60±0.16	1	1.05±0.08	0.21±0.06	100
8.17±0.06	1.31±0.14	2	2.22±0.14	0.20±0.02	80
8.13±0.05	2.58±0.12	2	2.06±0.03	0.28±0.04	95
8.03±0.04	6.17±0.17	2	21.0±0.13	0.36±0.06	75
8.03±0.05	9.57±0.17	2	1.90±0.16	0.29±0.06	95
8.26±0.04	1.80±0.14	4	5.78±0.41	0.43±0.07	15
8.18±0.05	2.79±0.12	4	4.60±0.40	0.50±0.09	85
8.12±0.05	6.91±0.11	4	4.86±0.46	0.58±0.07	75
8.07±0.06	10.15±0.14	4	3.92±0.30	0.56±0.07	85
8.37±0.04	1.60±0.33	8	9.75±0.40	0.82±0.10	0
8.33±0.04	3.27±0.21	8	8.82±0.90	0.99±0.03	0
8.28±0.07	7.02±0.23	8	9.80±0.95	1.16±0.11	0
8.26±0.07	10,58±0.29	8	7.29±0.42	1.18±0.07	0

Additional information

No reliable studies on the acute toxicity of reaction mass of aluminium hydroxide and aluminium nitrate and aluminium sulphate to fish are available.Therefore, a weight of evidence approach is applied and data are read-across to various aluminium compounds based on an analogue approach.

Two short-term studies according to OECD guideline 203 and GLP are available for aluminium sulfate. Both tests were performed with Danio rerio. One test was static without pH adjustment. In this test an LC50 was found of 1 mg/l (dissolved Al) at a pH of approximately 5. The second test was semi-static with pH adjustment. The test substance concentrations were in a range of 62.5 to 1000 mg/l (nominal), this resulted in dissolved aluminum concentrations of 0.247 to 0.105 mg/l (the concentration decreased with increasing nominal concentrations). The pH was between 7.4 and 8.0. No effects were found in this test, therefore, LC50 is greater than 0.247 mg/l (dissolved Al), which was the maximum soluble concentration under the test conditons. In a non-GLP and non-guideline study (Gundersen ,1994) Oncorhynchus mykiss was exposed to AlCl₃at four different water hardness concentrations (10, 30, 80 and 120 mg/L as CaCO3) and two pH values (7.3 and 8.3). Additional humic acid was added to some test series. In this test the different parameters, pH, water hardness and humic acid, had hardly an effect on the determined LC50 values. For total Al they were found within a range of 3.75 to 6.17 mg/L without distinct correlation to parameters. The same was found for dissolved Al were LC50-values were in the range of 0.36 to 0.79 mg/L. The study is considered to be reliable.Aluminium sulphate was used in a non-GLP non-guideline study with Salmo salar by Roy & Campbell (1997). In this study fish were exposed to a nominal pH of 5 and some fish were additionally exposed to fulvic acid. Also in this study LC50-value in both experiments were in the same range of magnitude from 78.3 to 224.1 µg/L.