Aluminum chloride hydroxide sulfate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study with sufficient documentation and analysis

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

WAF method

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

WAF method

Principles of method if other than guideline:

WAF method used following principals outlined in OECD Testing of Difficult substances

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

No information available in the report on sampling method.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Two methods were employed to prepare the test solutions. WAFs were prepared by direct addition of test substance into standard medium NF EN ISO 6341 and OECD 202
- Differential loading: In one case the solutions were prepared as WAFs, shaken manually and allowed to stand for a period of 4 days prior to use. Test solutions were prepared by directly pipetting from the centre of the vessel (type undisclosed) without disturbing the precipitarte hen present. In the second case solutions were prepared as WAFs, shaken and then filtered using a Millipore Durapore 0.45µm filter and the filtrate transferred to the test vessels.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): No dispersant or solvent used
- Concentration of vehicle in test medium (stock solution and final test solution): Nominal WAFs in both solutions were 0, 12.5, 25, 50, 100 and 200 mg/l.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): In the case of the unfiltered solutions, at 100 and 200 mg/l
precipitate was noted at the botom of the preparation vessels prior to use in the study

Test organisms (species):

Daphnia magna

Details on test organisms:

- Strain: Clone 5
- Source: not specified
- Age at study initiation (mean and range, SD): <24 h
- Feeding: raised on Elendt Shneider (M4) medium and fed on Pseudokirchneriella subcapitata

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

None

Hardness:

Not measured

Test temperature:

Between 18 and 20°C but actual measured temperature not specified in the report

pH:

7.25 - 7.77 in filtered test solutions
7.00 - 7.77 in decanted test solutions

Dissolved oxygen:

8.6-9.0 mg/l in filtered test solutions
8.6 - 9.0 mg/l in unfiltered test solutions

Nominal and measured concentrations:

0, 12.5, 25, 50, 100, 200 mg/l Nominal
Control not measured
See table below
The results from the final measured concentrations of unfiltered (and probably also the filtered) WAF suggest that the T0 and T48 hour results were inverted between sampling and analysis as they are inconsistent with observations and test substance characterisitics. However this is not thought to have effected the outcome or validity of the study

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Penicilin flasks
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water
- Total organic carbon: not measured
- Particulate matter: not measured
- Ca/mg ratio: As specified in Guideline
- Conductivity: not measured
- Culture medium different from test medium: M4
- Intervals of water quality measurement: 48 h (beginning and end of study)

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: performed in the dark

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 200 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 0
- Effect concentrations exceeding solubility of substance in test medium: At 200 mg/l 10% immobilisation was noted in the filtered and 5% in the unfilered solutions after 48 h

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- 24 h EC50: 1.6 mg/l
- Other: using Potassium dichromate

Reported statistics and error estimates:

None used due to immobilisation exceeding the maximum WAF loading used

Validity criteria fulfilled:

yes

Conclusions:

While a number of details are not available and the analytical data only makes sense when the values from T0 and T48 are inversed, this does not impact the validity of the study as effects were greater than the maximum loading rate of the test substance used and thus greater than the solubilkity limit of the dissolved aluminium concentration measured in the study.

Executive summary:

While a number of details are not available and the analytical data only makes sense when the values from T0 and T48 are inversed, this does not impact the validity of the study as effects were greater than the maximum loading rate of the test substance used and thus greater than the solubilkity limit of the dissolved aluminium concentration measured in the study. EC50 > 200 mg/L.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

20-06-1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Although the method is clear, the analytical data are questionable. the conclusion (text) from the range finding test is not in line with the tested concentrations. no test dates of the limit test are included. a high variation in actual concentrations in the limit test at the same nominal concentration is also questionable (from serial dilution). The conclusion therefore is that the results from this study are considered reliable with restrictions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

A sample of 4 ml was taken from the test solution at the beginning of the limit test and frozen. A further sample totalling 4 ml was taken from the test vessel containing daphnids at T24 hours nad at the end of the test and acidified using 50 uL HNO3 (analytical grade 99.5%)

Vehicle:

no

Details on test solutions:

The formulation was duissolved in M7 reconstituted water at 100 mg/L. the formulation was disperserd by agitating for 1 hour +- 6min and the resulting stock solution centrifuged at 2000 RPM for 15 minutes to remove any particles of undissolved formulation, the supernatant drawn off and used as the stock solution.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: CIT
- Age at study initiation (mean and range, SD): 6-24h

ACCLIMATION
- Acclimation period: none

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

386-418 mg/L

Test temperature:

19.3-19.8 C

pH:

7.68-7.76

Dissolved oxygen:

8.7-8.6 mg/L

Nominal and measured concentrations:

nominal: 100 mg/L
measured Al 3+: 0.15 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Aeration: no
- No. of organisms per vessel: 1 daphnid per 10 ml test solution

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.15 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

dissolved

Remarks:

aluminium

Basis for effect:

mobility

Analyisis confirms Al3+ concentration, no effects were observed, although 1 daphnid was foud to be dead (not significant). Analysis report is not reliable due to anomalies in date and test concentration of range finding test. and deviation in limit actual test concentrations.

Validity criteria fulfilled:

not specified

Conclusions:

In the limit test, the NOEC was determined to be 100 mg/L nominal and 0.15 mg/L for the measured dissolved Al3+

Executive summary:

Toxicity of AQUALENCE F to Daphnia magna was tested according to the OECD 202 guideline in a limit test. the nominal test concentration was 100 mg/L, analysis showed 0.07 mg/L Al3+ as backgroud noise. No toxicological effects were observed. The NOEC was determniend to be 100 mg/L nominal and 0.15 mg/L actual Al3 +. Expert judgement states that the NOEC should be 0.22 mg/L actual Al3 +. Analysis was performed but is very questionable. Concentrations on the analysis report do not match test concentrations as well as test date (range finding). the measured concentrations in the limit test have a high variance (around a factor 3), the conclusion from the range finding test also is questionable (since 0.1, 1.0 and 10mg/L were tested and the conclusion from the range finding is that the NOEC here is 100 mg/L).

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17-06-2004 to 12-08-2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: method is clear, however, results should be handeled with care

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

test water according to OECD 203

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Water phase samples were taken from the freshly prepared test solutions before filling
the individual beakers and from old media after decanting and pooling the water
per concentration by 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 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:

For each test item concentration, adequate amounts of the test item were mixed thoroughly
with reconstituted water in volumetric glass ware. After one hour, the pH was
adjusted to 7.5 by using NaOH. The prepared test liquid was sampled for chemical
analysis and filled into the test beakers. The daphnids were introduced directly thereafter.
The nominal test concentrations were 10, 20, 40, 80, and 160 mg/L. Reconstituted
water was tested as control.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna STRAUS, Crustacea, Cladocera (clone V).
Age: 4 - 24 hours old.
Origin: Umweltbundesamt, Institut für Wasser-, Boden- und Lufthygiene,
bred in the laboratory of the Fraunhofer-IME.
Breeding and holding conditions:
Adult Daphnia, at least 3 weeks old, were separated from the stock population by
sieving. Batches of 30 to 50 animals were held at room temperature in ca. 1800 mL
purified drinking water. During the week the daphnids are fed daily with an algal suspension
(Scenedesmus subspicatus) and LiquizellR (HOBBY) according to the EECGuideline
(ref 2). Algae growing in the log-phase, are centrifuged and the pellet is resuspended
in a few mL of medium. 30 mL of this suspension is given to 1 L Daphnia
medium. The water was changed once per week. Newborn daphnids were separated
by sieving, the first generation was discarded.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 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:

20.0-20.1 C

pH:

7.4-7.9

Dissolved oxygen:

93-98% of saturation

Nominal and measured concentrations:

nominal test substance: 10, 20, 40, 80, 160 mg/L
nominal aluminium: 0.907, 1.81, 3.63, 7.26, 14.5 mg/L
mean measured total Al: control (0.238), 0.614, 1.47, 3.20, 6.38, 14.2 mg/L
mean measured dissolved:

Details on test conditions:

Daphnids (Daphnia magna) were exposed to 5 concentrations of the test item in 4
replicates under semi-static conditions for a period of 48 hours. 60 mL glass beakers
were used as test vessels. To each beaker 50 mL test solution and 5 daphnids, not
older than 24 hours, were added. They were transferred to freshly prepared test liquids
in new test vessels after 24 h. No feeding and no aeration occurred throughout
the test. The controls were kept under the same conditions in reconstituted water.
Immobility and abnormal behaviour were recorded after 24 h and 48 h. Immobile animals
were eliminated from the vessels as soon as they were discovered. The daphnids
were considered to be immobile if they were not able to swim within 15 seconds
after gentle agitation of the test vessels. The temperature during the test was 20.0 ±
0.1 °C. The beakers were covered with glass panes and subjected to a light/dark cycle
of 16/8 h with light intensities of less than 1000 Lux.
In the freshly prepared test liquids at test start and after 24 h (before adding the
daphnids), and in the old liquids after 24 h and at test end pH-values (WTW Microprocessor
pH-Meter pH 196) and oxygen concentrations (WTW Microprocessor Oximeter
OXI 196) of the decanted and pooled test solutions and control water were
measured.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

98 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 77-126 mg/L

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

42 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

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 57 % of the lowest concentration analysed
in the unfiltered samples (total aluminium) and was in the range of the amounts
measured for the membrane-filtered samples (dissolved aluminium). Blank solutions
prepared from purified water showed no such background.
For the lowest concentration level the mean measured total aluminium concentration
was only 68 % of the nominal concentration level. For all concentrations above
10 mg/L mean measured total aluminium concentrations ranged between 81 and 98
% (Table 2). Since all values being relevant for the determination of (no) effect concentrations
were between dose levels of 20 and 160 mg/L, the determinations were
based on nominal concentrations.

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.
EC10 and EC50 values were determined together with 95% confidence intervals using
Probit-analysis assuming log-normal distribution of the values by using the ToxRat
programme. The NOEC was estimated as the highest concentration with immobility according to
the validity criterion for the controls (immobility not exceeding 10%).

Nominal contration (mg.L)	Cumulative immobility 24h				Cumulative immobility 48h
	No1	2	3	4	1	2	3	4
Control	0	0	0	0	0	0	0	0
10	0	0	0	0	0	0	0	0
20	0	0	0	0	0	0	0	0
40	0	0	1	1	0	0	1	1
80	3	1	1	2	3	2	1	2
160	4	3	2	3	4	4	3	4

Validity criteria fulfilled:

yes

Conclusions:

The 48h EC50 and EC10 are determined to be 98 and 42 mg/L nominal, respectively.

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

Daphnia magna.

Gilufloc 40 H (Polyaluminiumchloride) was tested at nominal concentrations of 10, 20,

40, 80, and 160 mg/L under semi-static conditions (daily renewal of the test medium)

for 48 hours. Test water was reconstituted water according to OECD TG 203, with pH

adjusted to 7.5 at each renewal.

The mean measured concentrations of total aluminium differed by less than 20 %

from the nominal values except the lowest concentration (68 % of the nominal concentration

of 10 mg/L, not relevant for the determination of (no) effect concentrations).

The concentrations of dissolved aluminium were in the range of the spectroscopic

background of the test medium. After subtraction of the background, all mean dissolved

Al concentration were < 0.1 mg/L, irrespective of the applied amount of total

aluminium.

However, the immobility of daphnids indicates a clear concentration-response relationship

when based on the test item. The resulting EC50 of the test item was determined

to be

EC50 = 98 mg test item/L (95 % confidence limits: 77 – 126 mg/L).

Thus it is concluded that toxicity is driven by other causes than dissolved aluminium.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17-06-2004 to 12-08-2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

restrictions involve the appearend dose response relation where no effect could be described by Al content but still endpoints regarding Al exposure are given based on this. It is therefore advised to use the EC50 given in the conclusion of this summary

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

test water was reconstituted according to OECD203

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

At test start water phase samples were taken from the test solution before filling the
individual beakers and at test end after 48 h after decanting and pooling the water
per concentration by 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 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:

For each test item concentration, adequate amounts of the test item were mixed thoroughly
with reconstituted water in volumetric glass ware. The prepared test liquid was
sampled for chemical analysis and filled into the test beakers. The daphnids were introduced
directly thereafter. The nominal test concentrations were 10, 20, 40, 80, and
160 mg/L. Reconstituted water was tested as control.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna STRAUS, Crustacea, Cladocera (clone V).
Age: 4 - 24 hours old.
Origin: Umweltbundesamt, Institut für Wasser-, Boden- und Lufthygiene,
bred in the laboratory of the Fraunhofer IME.

Breeding and holding conditions:
Adult Daphnia, at least 3 weeks old, were separated from the stock population by
sieving. Batches of 30 to 50 animals were held at room temperature in approx. 1800
mL purified drinking water. During the week the daphnids are fed daily with an algal
suspension (Scenedesmus subspicatus) and LiquizellR (HOBBY) according to the
EEC-Guideline (ref 2). Algae growing in the log-phase, are centrifuged and the pellet
is resuspended in a few mL of medium. 30 mL of this suspension is given to 1 L
Daphnia medium. The water was changed once per week. Newborn daphnids were
separated by sieving, the first generation was discarded.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 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:

20.0-20.1 C

pH:

5.1-8.0

Dissolved oxygen:

93-96% of saturation

Nominal and measured concentrations:

nominal test item: 10.0, 20.0, 40.0, 80.0, 160 mg/L
nominal aluminium: 0.907, 1.81, 3.63, 7.26, 14.5 mg/L

Details on test conditions:

Daphnids (Daphnia magna) were exposed to 5 concentrations of the test item in 4
replicates under static conditions for a period of 48 hours. 60 mL glass beakers were
used as test vessels. To each beaker 50 mL test solution and 5 daphnids, not older
than 24 hours, were added. No feeding and no aeration occurred throughout the test.
The controls were kept under the same conditions in reconstituted water. Immobility
and abnormal behaviour were recorded after 24 h and 48 h. Immobile animals were
eliminated from the vessels as soon as they were discovered. The daphnids were
considered to be immobile if they were not able to swim within 15 seconds after gentle
agitation of the test vessels. The temperature during the test was 20.0 ± 0.1 °C.
The beakers were covered with glass panes and subjected to a light/dark cycle of
16/8 h with light intensities of less than 1000 Lux.
At test start before adding the daphnids, and at test end pH-values (WTW Microprocessor
pH-Meter pH 196) and oxygen concentrations (WTW Microprocessor Oximeter
OXI 196) of the decanted and pooled test solutions and control water were measured.

Reference substance (positive control):

yes

Remarks:

pottasiumdichromate

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

21.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

38 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.214 - 1.26 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Remarks:

aluminium

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.45 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: total Al

Basis for effect:

mobility

Details on results:

The test medium concentrations are given for the test item Gilufloc 40 H
(Polyaluminiumchloride) with a certified Al2O3-content of 17.13%. This corresponds to
an Al-content of 9.07 %. The nominal Al content was calculated by using this value
(factor 0.0907).
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 the pH value after test item addition). The background
corresponded to a maximum value of 25 % of the lowest concentration analysed
in the unfiltered samples (total aluminium) and was in the range of the amounts
measured for the membrane-filtered samples (dissolved aluminium). Blank solutions
prepared from purified water showed no such background.
In the freshly prepared test media Al concentrations were in the range of 82 – 96 % of
the nominal values and deviated by less than 20 % from the nominal value. Since the
results of this study should be related to initial concentrations, nominal concentrations
could be taken as the basis for the determination of effect concentrations (ref 1).
Up to and including the nominal concentration of 80 mg/L, the Al-concentrations were
relatively constant after standing for 48 h (range 77 % - 91 %). At the highest concentration
of nominally 160 mg/L, the concentration decreased by one third after 48 h.
The 0.45 μm-filtered fraction, defined as dissolved aluminium was significantly
lower than the total aluminium fraction. After subtraction of the control background,
for all nominal concentrations up to and including 80 mg test item/L dissolved
Al values were < 0.1 mg/L irrespective of the applied amount of total Al.
The pH of these solutions were between 6.7 and 8.0.
Since the pH-values in the test media were not controlled, the dissolution of high
amounts of the test item lowered the pH of the solution, and the solubility of
Al was increased: At a pH of 5.1 (nominal concentration of 160 mg test item/L), the
dissolved aluminium concentration was approx. 1.3 – 1.4 mg/L, representing about 9 -
15 % of the total aluminium. Furthermore, at that concentration, the pH difference between
study start and end was only low (no changes of pH due to exceeded buffer
capacity of the medium).

Results with reference substance (positive control):

results are inline with guideline

Reported statistics and error estimates:

The evaluation of the effect concentrations was based on initially nominal test item
concentrations, since the initial test item concentrations, calculated from the initially
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 initially measured concentrations.
EC10 and EC50 values were determined together with 95% confidence intervals using
Probit-analysis assuming log-normal distribution of the values by using the ToxRat
programme. Due to the clear results, NOEC and LOEC could be estimated without statistical tests.

Since no aluminium was observed at nominal test substance concentration above the LOD, effects could not have been caused by aluminium. Therefore the EC50 suggested by the authors could not be supporten. In the controlgroup, however, aluminium was observed and in the highest concentration Al was observed. Expert judgement therefore states the EC50 based on measured free Al to be between 0.212 and 1.26 mg/L.

cumulative mortality:

Nominal concentration (mg/L)	24h				48h
	No1	2	3	4	1	2	3	4
Control	0	0	1	0	0	0	1	0
10	0	0	0	0	0	0	0	0
20	0	0	0	0	0	0	0	1
40	3	2	3	3	4	3	3	4
80	5	4	4	4	5	5	4	4
160	5	5	5	5	5	5	5	5

Validity criteria fulfilled:

yes

Conclusions:

Since no aluminium was observed at nominal test substance concentration above the LOD, effects could not have been caused by aluminium. Therefore the EC50 suggested by the authors could not be supporten. In the controlgroup, however, aluminium was observed and in the highest concentration Al was observed. Expert judgement therefore states the EC50 based on measured free Al to be between 0.212 and 1.26 mg/L.
Reported EC50 for the test compound was 38.0 mg/L nominal, EC10 was 21.1 mg/L nominal

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 Daphnia magna relevant for an accidental exposure. Gilufloc 40 H (Polyaluminiumchloride) was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under static conditions for 48 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. The initially measured concentrations based on total aluminium ranged from 82 % to 96 % of nominal values. The concentrations of dissolved Al were smaller than 0.1 mg/L irrespective of the nominal concentration, if the pH was in the range from 6.7 and 8.0. At the highest test concentration, the mean measured dissolved aluminium concentrations increased to 1.4 mg/L due to pH which was decreased to 5.1 by the test item. The EC50 of the test item was determined to be 38 mg test item /L (95 % confidence limits: 31 – 47 mg/L).

Since no aluminium was observed at nominal test substance concentration above the LOD, effects could not have been caused by aluminium. Therefore the EC50 suggested by the authors could not be supporten. In the controlgroup, however, aluminium was observed and in the highest concentration Al was observed. Expert judgement therefore states the EC50 based on measured free Al to be between 0.212 and 1.26 mg/L.

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Study period:

06-05-2003 tom 08-02-2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study is complete

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

reconstitution water according to the OECD 203 guideline

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Water phase samples were taken from the freshly prepared test solutions before filling
the individual beakers and from old media after decanting and pooling the water
per concentration by 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):

Daphnia magna

Details on test organisms:

Justification for the use of the test organism:
Daphnia magna (Crustacea, Phyllopoda, Cladocera) was chosen by OECD-experts (ref 1)
as test organism representing aquatic invertebrates.
8.1.2 Specification:
Species: Daphnia magna STRAUS, Crustacea, Cladocera (clone V).
Age: 4 - 24 hours old.
Origin: Umweltbundesamt, Institut für Wasser-, Boden- und Lufthygiene,
bred in the laboratory of the Fraunhofer-IME.
8.1.3 Breeding and holding conditions:
Adult Daphnia, at least 3 weeks old, were separated from the stock population by
sieving. Batches of 30 to 50 animals were held at room temperature in ca. 1800 mL
purified drinking water. During the week the daphnids are fed daily with an algal suspension
(Scenedesmus subspicatus) and LiquizellR (HOBBY) according to the EECGuideline
(ref 2). Algae growing in the log-phase, are centrifuged and the pellet is resuspended
in a few mL of medium. 30 mL of this suspension is given to 1 L Daphnia
medium. The water was changed once per week. Newborn daphnids were separated
by sieving, the first generation was discarded.
8.1.4 Sensitivity
The sensitivity of the test clone was checked by using K2Cr2O7 as reference substance.
Between 04.04.2003 and 13.05.2003 the EC50 ranged from 1.0 to 1.3 mg/L.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.0-20.1 C

pH:

7.5-8.0

Dissolved oxygen:

94-100% saturation

Nominal and measured concentrations:

nominal: 10, 20, 40, 80, 160 mg/L
nominal AL: 0.9, 1.8, 3.6, 7.2, 14.4 mg/L
measured Al: 1.11, 1.89, 4.02, 7.38, 14.1 mg/L (geomean)
measured AL (dissolved):0.172, 0.176, 0.166, 0.107, 0.115 mg/L (geomean)

Details on test conditions:

Daphnids (Daphnia magna) were exposed to 5 concentrations of the test item in 4
replicates under semi-static conditions for a period of 48 hours. 60 mL glass beakers
were used as test vessels. To each beaker 50 mL test solution and 5 daphnids, not
older than 24 hours, were added. They were transferred to freshly prepared test liquids
in new test vessels after 24 h. No feeding and no aeration occurred throughout
the test. The controls were kept under the same conditions in reconstituted water.
Immobility and abnormal behaviour were recorded after 24 h and 48 h. Immobile
animals were eliminated from the vessels as soon as they were discovered. The
daphnids were considered to be immobile if they were not able to swim within 15
seconds after gentle agitation of the test vessels. The temperature during the test was
20.0 ± 0.1 °C. The beakers were covered with glass panes and subjected to a
light/dark cycle of 16/8 h with light intensities of less than 1000 Lux.
In the freshly prepared test liquids at test start and after 24 h (before adding the
daphnids), and in the old liquids after 24 h and at test end pH-values (WTW Microprocessor
pH-Meter pH 196) and oxygen concentrations (WTW Microprocessor Oximeter
OXI 196) of the decanted and pooled test solutions and control water were
measured.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 160 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

measured concentrations appeared to be very pH dependent and reached maximum solubility. therefore the nominal and measured (dissolved) concentration) are very diffrent. The result therefore is that the NOEC exceeds the water solubility at the tested pH's.

Validity criteria fulfilled:

yes

Conclusions:

No signs of immobility wre observed during the 48h test period nor other signs of intoxication could be observed. the conclusion is that the NOEC >= 160 mg/L nominal and the NOEC exceeds the water solubility at the tested pH levels.

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 Daphnia magna. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under semi-static conditions (daily renewal of the test medium) for 48 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted to 7.5 at each renewal. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values for all concentrations except the lowest one. The concentrations of dissolved aluminium were measured between 0.1 and 0.2 mg/L, irrespective of the applied amount of total aluminium. Up to the highest test concentration, no immobility could be observed after 48 h. Thus, the NOEC of aluminium sulphate-14-hydrate under the described study conditions was determined as follows:

NOEC ≥ 160 mg aluminium sulphate-14-hydrate / L and exceed the water solubility at the tested pH levels.

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: All data is available. however, method of determining endpoints is questionable

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

At test start water phase samples were taken from the test solution before filling the
individual beakers and at test end after 48 h after decanting and pooling the water per
concentration by 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 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 μmmembrane
filtered samples generally are assumed to contain the “dissolved metal” fractions

Vehicle:

no

Details on test solutions:

For each test item concentration, adequate amounts of the test item were mixed
thoroughly with reconstituted water in volumetric glass ware. The prepared test liquid
was sampled for chemical analysis and filled into the test beakers. The daphnids
were introduced directly thereafter. The nominal test concentrations were 10, 20, 40,
80, and 160 mg/L. Reconstituted water was tested as control.

Test organisms (species):

Daphnia magna

Details on test organisms:

Specification:
Species: Daphnia magna STRAUS, Crustacea, Cladocera (clone V).
Age: 4 - 24 hours old.
Origin: Umweltbundesamt, Institut für Wasser-, Boden- und Lufthygiene,
bred in the laboratory of the Fraunhofer-IME.
Breeding and holding conditions:
Adult Daphnia, at least 3 weeks old, were separated from the stock population by
sieving. Batches of 30 to 50 animals were held at room temperature in approx. 1800
mL purified drinking water. During the week the daphnids are fed daily with an algal
suspension (Scenedesmus subspicatus) and LiquizellR (HOBBY) according to the
EEC-Guideline. Algae growing in the log-phase, are centrifuged and the pellet
is resuspended in a few mL of medium. 30 mL of this suspension is given to 1 L
Daphnia medium. The water was changed once per week. Newborn daphnids were
separated by sieving, the first generation was discarded.
Sensitivity
The sensitivity of the test clone was checked by using K2Cr2O7 as reference substance.
Between 04.04.2003 and 13.05.2003 the EC50 ranged from 1.0 to 1.3 mg/L.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 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:

20.0-20.1 C

pH:

4.6-8.0 decreasing pH with increasing Al concentration

Dissolved oxygen:

91-97% of saturation

Nominal and measured concentrations:

nominal testsubstance: 10.0, 20.0, 40.0, 80.0, 160 mg/L
nominal aluminium: 0.900, 1.80, 3.60, 7.20, 14.4 mg/L
measured aluminium in test solution begin: 0.078, 0.039, 0.029, 0.468, 5.00

Reference substance (positive control):

yes

Remarks:

Pottasium dichromate

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

30.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

47.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 40.5-55.6 mg/L

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

2.8 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

other: total aluminium

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

4.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

other: total aluminium

Basis for effect:

mobility

Remarks on result:

other: 3.6-5.0

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.33 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

dissolved

Remarks:

aluminium

Basis for effect:

mobility

Results with reference substance (positive control):

EC50 from reference substance test was 1.0 -1.3 mg/L

Reported statistics and error estimates:

The evaluation of the effect concentrations was based on initially nominal test item
concentrations, since the initial test item concentrations, calculated from the initially
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 initially measured concentrations.
EC10 and EC50 values were determined together with 95% confidence intervals using
Probit-analysis assuming log-normal distribution of the values by using the ToxRat
programme

Nominal concentration (mg/L)	24h				48h
	No. 1	2	3	4	1	2	3	4
Control	0	0	0	0	0	0	0	0
10	0	0	0	0	0	0	0	0
20	0	0	0	0	0	0	1	0
40	0	0	0	0	0	0	3	0
80	1	2	4	3	5	5	5	5
160	5	5	5	5	5	5	5	5

Validity criteria fulfilled:

yes

Conclusions:

The EC50 of the test item was determined to be 48 mg test item /L (95 % confidence limits: 41 – 56 mg/L) nominal. and 0.33 mg/L bioavailable aluminium. the EC10 was determined to be 30.8 mg/L nominal. Endpoint determinations are questionable, therefore NOEC/LOEC data are not to be used. EC50 of the bioavailable concentration of aluminium was calculated in a very questionable way but adaptations would lead to minor differenes in EC50. Therefore this value still seems 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 aluminium sulphate-14-hydrate to Daphnia magna relevant for an accidental exposure. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under static conditions for 48 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. The initially measured concentrations based on total aluminium differed by less than 20 % from nominal values. The mean measured dissolved aluminium concentrations strongly depended on pH which was influenced by the test item. The EC50 of the test item was determined to be - 48 mg test item /L (95 % confidence limits: 41 – 56 mg/L). The concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration: 48 mg/L of aluminium sulphate-14-hydrate reduced the pH of reconstituted water (OECD 203) to a level which enabled 0.33 mg Al/L to be dissolved, causing 50 % immobility after 48 h.

Description of key information

OECD guideline 202, GLP, key study, validity 1 (Thiebaud, 2000, static test), registered substance : aluminium chloride hydroxide sulfate

48h-EC50 > 200 mg/L (based on nominal concentrations)

OECD guideline 202, GLP, read-across study, validity 2 (Schafers, 2004, static test), read-across aluminium trichloride

48h-EC50 = 0.214 - 1.26 mg/L (based on measured intial concentrations, dissolved aluminium)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

200 mg/L

Additional information

Two studies were performed on aluminium chloride hydroxide sulphate to evaluate its short-term toxicity to aquatic invertebrates.

1) While a number of details are not available and the analytical data only makes sense when the values from T0 and T48 are inversed, this does not impact the validity of the study as effects were greater than the maximum loading rate of the test substance used and thus greater than the solubility limit of the dissolved aluminium concentration measured in the study. EC50 > 200 mg/L.

2) Toxicity of AQUALENCE F to Daphnia magna was tested according to the OECD 202 guideline in a limit test. the nominal test concentration was 100 mg/L, analysis showed 0.07 mg/L Al3+ as backgroud noise. No toxicological effects were observed. The NOEC was determniend to be 100 mg/L nominal and 0.15 mg/L actual Al3 +. Expert judgement states that the NOEC should be 0.22 mg/L actual Al3 +. Analysis was performed but is very questionable. Concentrations on the analysis report do not match test concentrations as well as test date (range finding). The measured concentrations in the limit test have a high variance (around a factor 3), the conclusion from the range finding test also is questionable (since 0.1, 1.0 and 10mg/L were tested and the conclusion from the range finding is that the NOEC here is 100 mg/L).

And 4 read across studies are available:

3) 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 Daphnia magna relevant for an accidental exposure. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under static conditions for 48 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. The initially measured concentrations based on total aluminium differed by less than 20 % from nominal values. The mean measured dissolved aluminium concentrations strongly depended on pH which was influenced by the test item. The EC50 of the test item was determined to be - 48 mg test item /L (95 % confidence limits: 41 – 56 mg/L). The concentration-effect relationship for the test item integrates the enhanced solubility of aluminium at the pH caused by the test item concentration: 48 mg/L of aluminium sulphate-14-hydrate reduced the pH of reconstituted water (OECD 203) to a level which enabled 0.33 mg Al/L to be dissolved, causing 50 % immobility after 48 h.

4) 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 Daphnia magna. Aluminium sulphate-14-hydrate was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under semi-static conditions (daily renewal of the test medium) for 48 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted to 7.5 at each renewal. The mean measured concentrations based on total aluminium differed by less than 20 % from nominal values for all concentrations except the lowest one. The concentrations of dissolved aluminium were measured between 0.1 and 0.2 mg/L, irrespective of the applied amount of total aluminium. Up to the highest test concentration, no immobility could be observed after 48 h. Thus, the NOEC of aluminium sulphate-14-hydrate under the described study conditions was determined as follows:

NOEC ≥ 160 mg aluminium sulphate-14-hydrate / L and exceed the water solubility at the tested pH levels.

5) 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 Daphnia magna relevant for an accidental exposure. Gilufloc 40 H (Polyaluminiumchloride) was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under static conditions for 48 hours. Test water was reconstituted water according to OECD TG 203, without pH adjustment. The initially measured concentrations based on total aluminium ranged from 82 % to 96 % of nominal values. The concentrations of dissolved Al were smaller than 0.1 mg/L irrespective of the nominal concentration, if the pH was in the range from 6.7 and 8.0. At the highest test concentration, the mean measured dissolved aluminium concentrations increased to 1.4 mg/L due to pH which was decreased to 5.1 by the test item. The EC50 of the test item was determined to be 38 mg test item /L (95 % confidence limits: 31 – 47 mg/L).

Since no aluminium was observed at nominal test substance concentration above the LOD, effects could not have been caused by aluminium. Therefore the EC50 suggested by the authors could not be supporten. In the controlgroup, however, aluminium was observed and in the highest concentration Al was observed. Expert judgement therefore states the EC50 based on measured free Al to be between 0.212 and 1.26 mg/L.

6) 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 Daphnia magna. Gilufloc 40 H (Polyaluminiumchloride) was tested at nominal concentrations of 10, 20, 40, 80, and 160 mg/L under semi-static conditions (daily renewal of the test medium) for 48 hours. Test water was reconstituted water according to OECD TG 203, with pH adjusted to 7.5 at each renewal.

The mean measured concentrations of total aluminium differed by less than 20 % from the nominal values except the lowest concentration (68 % of the nominal concentration

of 10 mg/L, not relevant for the determination of (no) effect concentrations). The concentrations of dissolved aluminium were in the range of the spectroscopic

background of the test medium. After subtraction of the background, all mean dissolved Al concentration were < 0.1 mg/L, irrespective of the applied amount of total

aluminium. However, the immobility of daphnids indicates a clear concentration-response relationship when based on the test item. The resulting EC50 of the test item was determined to be EC50 = 98 mg test item/L (95 % confidence limits: 77 – 126 mg/L). Thus it is concluded that toxicity is driven by other causes than dissolved aluminium.