A mixture of: sodium/potassium 7-[[[3-[[4-((2-hydroxy-naphthyl)azo)phenyl]azo]phenyl]sulfonyl]amino]-naphthalene-1,3-disulfonate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Remarks:

Reproduction of Daphnia magna

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 April 1994 to 19 July 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: "Daphnia sp., Acute Immobilisation Test and Reproduction Test"

Version / remarks:

Part II, adopted April 04, 1984.

Deviations:

not specified

Principles of method if other than guideline:

The OECD draft method used later on became adopted as OECD method 211

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Code number: FAT45155/B
Batch number: TV1, HT3728
Appearance: solid
Solubility: in water: 48 g//; in DMSO and DMF: soluble
Storage: at room temperature, in the dark
Expiration date: 15.5.1997

Details on sampling:

The analytically determined test substance concentrations in the stock solution samples varied in the range of 93.3 % to 110.0 % of the nominal values. The measured test substance concentrations in the samples from the freshly prepared test media varied in the range from 75.0 % to 114.1 % of the nominal values. In the samples, taken from the test media at the end of the renewal periods of 48 and 72 hours, concentrations were measured in the range from 70.0 % to 148.4 % of the nominal values. No significant decrease of the test substance concentrations were determined during the renewal periods of 48 to 72 hours. And also no clear difference was observable between the measured concentrations in the test media samples without respectively with algae. Thus, FAT 45155/B was obviously sufficiently stable under the test conditions at least over the longest renewal periods of 72 hours.

Vehicle:

no

Details on test solutions:

At each treatment a stock solution was freshly prepared just before each test medium renewal by dissolving the test substance into test water (100 mg/1). Then adequate volumes of the intensively stirred stock solution were mixed into test water to obtain the desired test concentrations before introduction of Daphnia.
The nominal concentrations tested were 0.10, 0.32, 1.0, 3.2 and 10.0 mg test substance/1. Additionally, a control was tested in parallel (test water without addition of the test substance).

Test organisms (species):

Daphnia magna

Details on test organisms:

The study has been performed with females of a clone of the species Daphnia magna Straus. The clone was bred in the laboratories of RCC under the same standardized conditions and in the same kind of test water as used in the test. At the start of the test the young Daphnia used were less than 24 hours old.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

2.5 mmol/L (approx. 250 mg/L as CaCo3)

Test temperature:

water temperature 20.5 - 21.4 °C during test period

pH:

pH 7.9 ±0.3

Dissolved oxygen:

before test the test water was aerated until oxygen saturation, whereas during the test the water was not aerated.

Salinity:

Alkalinity: 0.9 mmol/L

Nominal and measured concentrations:

For the analysis of the test substance concentrations, single samples were taken from the freshly prepared stock solutions and duplicate samples from the freshly prepared test media of all test concentrations and the control at the first treatment period (Day 0), at a treatment period in the second week (Day 12) and at the last treatment period (Day 16).

Details on test conditions:

In this semistatic test the test media of all test concentrations and of the control were renewed on Days 2, 5, 7, 9, 12, 14, 16 and 19 of the exposure period (every Monday, Wednesday and Friday). By that, a total of 9 treatments were performed. At these dates the test animals were carefully transferred by glass tubes from the old test vessels into the freshly prepared test media of the corresponding concentrations. The Daphnia were fed at the same time intervals and additionally on Tuesday and Thursday with green algae (Scenedesmus subspicatus), freshly grown in the laboratories of RCC.

The amounts of algae added:
Days 0/1/6/8/13/15: 50,000 cells/ml
Days 2/5/7/12/14/20: 150,000 cells/ml
Days 9/16/19: 200,000 cells/ml

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 0.8 mg/L measured

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: equivalent to 3.5 mg/L

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: equivalent to 0.8 mg/L measured

Details on results:

The LOEC for reproduction could not be determined as no animal survived until the end of the test at 3.2 and 10 mg/L nominal. With exception of the reported mortality and the reduced reproduction rates, no particular signs of intoxication were observed at the test animals during the test. Taking the survival rates and the reproduction rates of the test animals into account, the highest concentration of FAT 45155/B tested without toxic effects (NOEC) after the exposure period of 21 days was determined to be 0.8 mg test substance/L (as the mean measured test substance concentration at the nominal value of 1.0 mg/L). The lowest concentration tested with toxic effects (LOEC) was determined to be 3.5 mg test substance/L (as the mean measured test substance concentration at the nominal value of 3.2 mg/L) due to the death of all Daphnia magna in this test concentration during the 21 days of exposure.

Survival of adults

In the control and in the test concentrations up to and including nominal 1.0 mg test substance/l the survival rate of the test animals amounted to at least 90 % at the end of the test after 21 days of exposure. At the two highest test concentrations of nominal 3.2 and 10.0 mg test substance/l, all Daphnla were dead at Day 16 and Day 14, respectively. Thus, the survival rate of Daphnia magna after 21 days was significantly reduced first at the nominal test concentration of 3.2 mg/l (mean measured test substance concentration: 3.5 mg/l).

 

Reproduction rate

The first young Daphnia released from their parent animals were recorded in the control and in the test concentrations up to and including nominal 3.2 mg test substance/l at the observation on Day 9. First at the highest test concentration (nominal 10.0 mg/l) a delay of the first brood was determined, the first offspring were observed on Day 12 of exposure. The mean reproduction rate of those Daphnia in the control was 87.7 ± 12.7 (mean ± SD) alive offspring per adult. No significant toxic effect of the test substance on the reproduction rate was determined up to and including the nominal test concentration of 1.0 mg test substance/l. At the test concentrations of nominal 3.2 and 10.0 mg/l the reproduction rate of Daphnia could not be calculated, since in these test concentrations no Daphnia survived until the end of test.

 

Signs of intoxication

With exception of the reported mortality and the reduced reproduction rates, no particular signs of intoxication were observed at the test animals during the test.

Validity criteria fulfilled:

yes

Conclusions:

The NOEC of FAT 45155/B in the daphnia magna reproduction toxicity test was determined at 1 mg/L (nominal) and the LOEC was 3.2 mg/L (nominal).

Executive summary:

In a GLP-compliant study, the influence of FAT 45155/B on the reproduction and survival rate of Daphnia magna was investigated in a 21-days semistatic test following the OECD Guideline 202. The nominal concentrations tested were 0.10, 0.32, 1.0, 3.2 and 10.0 mg test substance/l and a control. The analytically determined test substance concentrations in the freshly prepared test media varied in the range from 75.0 % to 114.1 % of the nominal values. At the end of the test medium renewal periods of 48 and 72 hours, concentrations were measured in the range from 70.0 % to 148.4 % of the nominal values. FAT 41017/A was obviously sufficiently stable under the test conditions at least over the longest renewal periods of 72 hours. Therefore, the mean values were calculated over all measured test substance concentrations (averages over all measurements at 0, 48 and 72 hours), and the biological results are related to these mean measured test substance concentrations. In the control and in the test concentrations up to and including nominal 1.0 mg test substance/l the survival rate of the test animals amounted to at least 90 % at the end of the test after 21 days of exposure. At the two highest test concentrations of nominal 3.2 and 10.0 mg test substance/l, all Daphnia were dead at Day 16 and Day 14, respectively. Thus, the survival rate of Daphnia magna after 21 days was significantly reduced first at the nominal test concentration of 3.2 mg/l (mean measured test substance concentration: 3.5 mg/l). The mean reproduction rate of those Daphnia in the control was 87.7 ± 12.7 (mean ± SD) alive offspring per adult. No significant toxic effect of the test substance on the reproduction rate was determined up to and including the nominal test concentration of 1.0 mg test substance/l. At the test concentrations of nominal 3.2 and 10.0 mg/l the reproduction rate of Daphnia could not be calculated, since in these test concentrations no Daphnia survived until the end of test. With exception of the reported mortality and the reduced reproduction rates, no particular signs of intoxication were observed at the test animals during the test. No remarkable observations were made concerning the behaviour of the test substance in the test media. At least the test media of nominal 0.32 to 10 mg/L were visibly coloured by the test substance. The biological test results after the exposure period of 21 days:

Highest concentration tested without toxic effects (NOEC): 0.8 mg test substance/l

Lowest concentration tested with toxic effects (LOEC): 3.5 mg test substance//

At the LOEC all Daphnia magna died during the 21 days of exposure.

Description of key information

NOEC (daphnia magna, 21 days, mortality & reproduction): 1.0 mg/L (nominal)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1 mg/L

Additional information

In a GLP-compliant study, the influence of FAT 45155/B on the reproduction and survival rate of Daphnia magna was investigated in a 21 days semistatic test following the OECD Guideline 202. The nominal concentrations tested were 0.10, 0.32, 1.0, 3.2 and 10.0 mg test substance/l and a control. In the control and in the test concentrations up to and including nominal 1.0 mg test substance/l the survival rate of the test animals amounted to at least 90 % at the end of the test after 21 days of exposure. At the two highest test concentrations of nominal 3.2 and 10.0 mg test substance/l, all Daphnia were dead at Day 16 and Day 14, respectively. Thus, the survival rate of Daphnia magna after 21 days was significantly reduced first at the nominal test concentration of 3.2 mg/l (mean measured test substance concentration: 3.5 mg/l). With exception of the reported mortality and the reduced reproduction rates, no particular signs of intoxication were observed at the test animals during the test. No remarkable observations were made concerning the behaviour of the test substance in the test media. At least the test media of nominal 0.32 to 10 mg/L were visibly coloured by the test substance. Under the conditions of this daphnia magna reproduction toxicity test the NOEC was determined at 1 mg/L (nominal/ 0.8 mg/L measured) and the LOEC at the next higher concentration of 3.2 mg/L (nominal/ 3.5 mg/L measured), showing 100 % mortality. At 1.0 mg/L (nominal) the reproduction rate was not affected compared to controls. Mortality at the 3.2 and 10 mg/L groups only began following day 7 and within one week all animals were found dead at these concentrations.