222 OP

Endpoint summary

Additional information

Short-term toxicity to fish

Two Key Studies according OECD Guideline 203 are available. In one study, the test item was found to cause no toxic effects to Zebrafish after 96 hours when tested with a limit concentration of 100 mg/L.

The results of the second study showed that under valid static test conditions, the 96 h-LC50 of the test item to fish (Rare minnow, Gobiocypris) is greater than the nominal concentration of 100 mg/L (measured concentration 95.1 mg/L), while the maximum concentration causing no mortality (96 h-LC0) is equal to the nominal concentration of 100 mg/L (measured concentration 95.1 mg/L). i.e.:

Therefore, the LC50 after 96 hours was > 100 mg/L on both studies. All effect levels are given based on the nominal concentration level of the test item.

Long-term toxicity to fish

Under semi-static conditions of 72 h-renewal, the effect on growth rates and other observed effectsin juvenile fish (Gobiocypris rarus) exposed to the test substance for 28 days was conducted according to the following guidelines:“The guidelines for the testing of chemicals”,SEPA(HJ/T 153-2004);Procedure 215of the“Guidelines for Testing of Chemicals”of the OECD: " Fish, Juvenile Growth Test” (2000)etc.

During the whole test period, the pH values of the control and Test Media were between 5.12 and 7.47, and the Dissolved Oxygen (DO) values varied from 85% to 92% of the air saturation at the test temperature, and the temperature of the Test Media was maintained in 23.0°C~5°C. All fishes in the control group were normal. The mean weight of fish in the controls increased more than 50%. So the study met the acceptability criteria prescribed by the protocol (dissolved oxygen concentration: no less than 60% of the air saturation value; temperature:(23±2)^oC andnot differ by more than ±1^oC between test chambers; the increasing rate of the mean wet-weight of fish: no less than 50% of the initial weight). Therefore the test was considered valid.

The concentration of the test item in test medium was determined by ICP-MS to represent the stability of the test substance in test period. A linear regression equation was obtained with the ICP-MS response values of phosphorus (y) vs. the concentration of the test item(mg/L):A=1172.4c+237.03, with good linearity of R=0.989. The results show that linearity for the concentration range of 0.02 mg/L to 0.50 mg/L is good. Besides, the minimum detection amount of ICP-MSfor the test item is 1.0×10^-10g, and the minimum detection concentration for water sample is 0.01mg/L. The analytical results showed that the concentration of the test item is consistent in the test medium throughout a period of 72 h (deviation within 20%). Thus a semi-static test of 72 h-renewal was reasonable.

The results showed that under valid semi-static test conditions (72 h-renewal), the LOEC (lowest observed effect concentration) and the NOEC (No observed effect concentration) of the test substance to juvenile Rare minnow were concluded as follows:

28 d-LOEC > 100 mg/L(measured concentration 96.9 mg/L);

28 d-NOEC = 100 mg/L(measured concentration 96.9 mg/L).

Short-term toxicity to daphnia

In the acute immobilization test with Daphnia magna (STRAUS) the effects test item at the limit test item concentration of 100 mg/L were determined according to OECD 202 (2004). The limit test was conducted under semi-static conditions over a period of 48 h.

Twenty daphnids were exposed to the limit concentration and the control. The Al³⁺content, component of the test item, was analytically verified with a cuvette test (cuvette test no. 1-02, Macherey-Nagel) by spectrophotometry in the fresh media (0 and 24 h) and the old media (24 and 48 h) of the limit concentration and the control.

The measured Al³⁺concentrations in the fresh media (0 and 24 h) and in the old media (24 and 48 h) were 102 % of the nominal value. All effect values given are based on the nominal concentration of the test item.

Water quality parameters (pH-value and dissolved oxygen concentration), measured in the fresh (0 and 24 h) and old media (24 and 48 h), were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

EC-Values, NOEC and LOEC

(based on the nominal concentration of the limit test item concentration)

	Test Duration [h]	Nominal Concentration of the Test Item [mg/L]	Confidence Interval p = 95% [mg/L]
EC10/ EC50/ EC100	24	> 100	not applicable
	48	> 100	not applicable
NOEC	48	100	---
LOEC	48	> 100	---

At the limit concentration of the test item of 100 mg/L, no effects were observed on Daphnia magna.

Long-term toxicity to daphnia

The Daphnia magna Reproduction Test (Semi-Static, 21 d) was conducted as a limit test with the limit concentration of 100 mg/L of the test item according to OECD 211 (2008).

Test species was Daphnia magna STRAUS (Clone 5). Ten daphnids individually held were used for the limit concentration and the control. At the start of the exposure, the daphnids were 2 to 24 h old. The test method was semi-static with renewal of the test solutions three times per week. The aim of the Daphnia Reproduction Test over 21 days was to assess effects on the reproduction capacity and other test item-related effects on parameters such as intrinsic rate of natural increase, occurrence of aborted eggs and stillborn juveniles, time of production of the first brood, adult mortality, dry body weight and length of the parental daphnids.

The content of aluminium of the control and the test item suspension with 100 mg/L substance was analytically verified with a validated photometric cuvette test on days 0, 9, 14 (fresh media, 0 hours), on days 2, 16 (old media, 48 hours) and on day 12 (old media, 72 hours).

The measured concentrations of the component aluminium in the fresh media (0 h) were the range of 68 to 85 % of the nominal value. The measured concentrations of the component aluminium in the old media (48 and 72 h) were the range of 85 to 99 % of the nominal value, which demonstrates that the test item was fully present throughout the exposure. The lack of the measured values in the fresh media may be explained by inhomogeneity of the test item dispersion.

The results given are based on the nominal concentration of 100 mg/L of the test item.

·  The averagevalues of living juveniles per parental daphnid were 82 for the control and 81 for the limit concentration of 100 mg/L after 21 days. The reproductive output of daphnids in the limit concentration was comparable to the reproductive output of daphnids in the control(t-test,p = 0.05).

·  The coefficient of variation of the number of living offspring produced per parent was 12 % for the control and 5 % for the limit concentration of 100 mg/L.

·  The intrinsic rates of natural increase (IR) of the surviving parent daphnids accounting for generation time and number of offspring were calculated as a measure for population growth and maintenance.The intrinsic rates of the limit concentration were comparable to those in the control(t-test,p = 0.05).

·  No dead juveniles were observed in control and the limit concentration of 100 mg/L. The number of aborted eggs was 4 in the control and 3 in the limit concentration. Related to the total number of produced juveniles (dead + alive) the percentage of dead juveniles and aborted eggs was 1 % at the limit concentration and the control.

·  Four broods were observed during the test period for all parental daphnids. The first brood was released from all parent daphnids in the limit concentration of 100 mg/L and in the control on days 8 and 9.

·  No adult mortality or immobilisation of parental daphnids was observedin the control and the limit concentration.

·  The mean dry body weight and total body lengths of all parental daphnids of the test and the control group were determined at the end of the study.
The mean dry body weight of the parental daphnids was 1.19 mg per daphnid for the control and 1.05 mg per daphnid for the limit concentration of 100 mg/L.
The mean body length of the parental daphnids was 5.23 mm per daphnid for the control and 5.05 mm per daphnid for the limit concentration.

·  No males and ephippia (winter eggs) were observed in the respective control or test groups.

No biologically significant effect could be determined in the limit concentration of100 mg/L in comparisonto the control.

The water quality parameters (pH-value, dissolved oxygen, water hardness and temperature) were within the acceptable limits.

·  In order to prove the validity of the test system and test conditions at the test facility, an acute immobilisation test according to DIN 38412 L 11 is carried out with potassium dichromate as reference item once per month.
The EC₅₀of the reference item at 1.73 mg/L (95% confidence limits: 1.67 - 1.79 mg/L) after 24 hours is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according toAQS P 9/2 (05/1996) for daphnids clone 5 cultured in Elendt M4 medium. The EC₅₀-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Test species was Daphnia magna STRAUS (Clone 5). Ten daphnids, individually held, were used per concentration level and control. At the test start, the daphnids were 2 to 24 hours old. The study was carried out under semi-static conditions with renewal of the test solutions three times per week. Aim of the Daphnia Reproduction Test over 21 days was to assess effects on the reproduction capacity and other test item-related effects or parameters such as time of production of first brood, adult mortality, intrinsic rate of natural increase, occurrence of aborted eggs and stillborn

         

Summary of all Test Item Related Effects

(Based on the nominal test item concentration of100 mg/L)

Effects	Control	Nominal Test Item Concentration 100 mg/L	Significance
Mean Number of Juveniles per Producing Parent (Reproduction Rate±SD)	82±9	81 ± 4	no^1)
Coefficient of Variation of theMean Number of Juveniles per Producing Parent	12	5	no
Intrinsic Rate of Natural Increase (Mean±SD)	0.44 ± 0.02	0.44 ± 0.02	no^1)
Number of Stillborn Juveniles and Aborted Eggs	1	1	no
Appearance of First Brood (Mean Day±SD)	8.5 ± 0.5	8.5 ± 0.5	no
Number of Broods	4	4	—
Adult Mortality after 21 Days [%]	0	0	no
Parent Daphnids: Mean Dry Weight [mg]	1.19	1.05	—
Parent Daphnids: Mean Body Length [mm]±SD	5.23 ± 0.25	5.05 ± 0.16	no^1)
NOEC	100 mg/L

Toxicity to algae

In this study the test item was found to inhibit the growth of the freshwater green alga Desmodesmus subspicatus after 72 hours. The nominal NOEC-values for inhibition of growth rate and inhibition of yield after 72 hours were 40.0 and 16.0 mg/L, corresponding to geometric mean measured values of 4.40 and 2.20 mg/L, respectively. The nominal LOEC-values for inhibition of growth rate and inhibition of yield after 72 hours were 100 and 40.0 mg/L, corresponding to 15.5 and 4.40 mg/L, respectively. The nominal EC50-values for inhibition of growth rate (ErC50) and yield (EyC50) with 95 % confidence intervals after 72 hours were > 100 and 59.5 (46.5 – 85.2) mg/L, corresponding to > 15.5 and 6.11 (4.90 – 10.4) mg/L, respectively.

All effect levels are given based on nominal and geometric mean measured concentrations of the test item.

Toxicity to microorganisms

A Respiration Inhibition Test with activated sludge according to OECD 209 was carried out with the substance under static conditions with the nominal substance concentrations 100 -320 -1000 mg/L. The respiration rates of the control, reference and test substance replicates were measured after a contact time of 3 h, and the inhibitory effects of the test and reference substance were determined in comparison to the control respiration rates. The mean inhibition of respiration for the test substance replicates ranged from 2 % to 24 %.

As a result of the test, the NOEC of the substance was determined to be 100 mg/L, the EC50 is > 1000 mg/L and the EC10 is 262 mg/L to activated sludge of a municipal sewage treatment plant.