Hydrogen bis[2-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]benzoato(2-)]chromate(1-), compound with 2-ethylhexylamine (1:1)

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen bis[2-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]benzoato(2-)]chromate(1-), compound with 2-ethylhexylamine (1:1)

EC Number:

275-864-7

EC Name:

Hydrogen bis[2-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]benzoato(2-)]chromate(1-), compound with 2-ethylhexylamine (1:1)

Cas Number:

71701-15-0

Molecular formula:

C42H44CrN9O6

IUPAC Name:

Chromate(1-), bis[2-[2-[4,5-dihydro-3-methyl-5-(oxo-kO)-1-phenyl-1H-pyrazol-4-yl]diazenyl-kN1]benzoato(2-)-kO]-, hydrogen, compd. with 2-ethyl-1-hexanamine (1:1:1)

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

Preparation of the saturated solution:
The saturated solution (1.00 mg/L test item were weighed out) was prepared with dilution water two days prior to the start of the exposure
(at -48 hours).

Treatment:
The test item was dispersed in the dilution water by stirring with approximately 1100 rpm for 48 hours at room temperature. After completion of
stirring, the test item dispersion was filtered with a membrane filter (0.45 µm, RC, MACHEREY-NAGEL) to achieve the saturated solution.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Clone 5
- Source:
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder: DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
- Age at study initiation (mean and range, SD): 2 to 24 h old daphnids from a healthy stock were used for the study
- Method of breeding:
Culture: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 +- 2°C, in an incubator, 16 h illumination; light intensity
of max. 20 µE x m-2 x s-1
Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
Cultur feeding: At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and
Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: At least 2 h in dilution water.
- Acclimation conditions (same as test or not): Same as test conditions.
- Health during acclimation (any mortality observed): No mortality observed.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

253 mgCaCO3/L

Test temperature:

18 - 22°C, constant within ± 1°C
At start of exposure: 19.7 °C

pH:

pH at the Start of the Exposure (0 hours): 7.84
at the end of the test:
Saturated solution: 7.72 - 7.83
Control: 7.78- 7.81

Nominal loading
of the test item
[mg/L] pH-value

1.00 6.91
Control 7.65


pH at the End of the Exposure (48 hours)
(measured in all replicates)

Nominal loading
of the test item
[mg/L] pH-values
Replicates
1 2 3 4
1.00 7.26 7.26 7.27 7.31
Control 7.13 7.14 7.19 7.22

Dissolved oxygen:

O2-concentration at the Start of the Exposure (0 hours): 8.20 mg O2/L
at the end of the test:
Saturated solution: 8.26 mg O2/L
control: 8.04 mg O2/L


Nominal loading level
of the test item
[mg/L]
Dissolved
O2-concentration
[mg/L]
1.00 4.38
Control 8.59



O2-concentration at the End of the Exposure (48 hours)
(measured in all replicates)

Nominaltest loading level
of the test item
[mg/L] Dissolved O2 -concentration [mg/L]
Replicates
1 2 3 4
1.00 8.41 8.38 8.42 8.44
Control 8.26 8.26 8.29 8.27

Nominal and measured concentrations:

nominal: 100 mg/L as a saturated solution (The maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, acc. to OECD Series, No. 23 (2000)6)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (5 cm ID x 8 cm H), 50 mL volume
- Test volume: 20 mL
- 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
- dilution water: According to EC 92/69 L383A C.2. Annex

- Culture medium different from test medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, illumination range max. 20 μmol photons⋅m-2 ⋅ s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Immobilisation, 24 h interval

TEST CONCENTRATIONS
-The effects of the saturated solution (100 mg/L) of the test item were tested in a limit test.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

other: 5 % Immobilisation

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Details on results:

See below

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC100-value was determined directly from the test results.
The EC10- and EC50- with 95 % confidence interval
(CI) was determined in a probability network by interpolation according to standard procedures.
EC-values after 24 h of the reference item in mg/L:
EC10 : 0.97
EC50 : 2.07 (CI 1.68 - 2.56)
EC100 : 5.80
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality
criteria according to AQS: DIN Guideline 38412 L 30.

Any other information on results incl. tables

Biological Data

The percentage immobility was determined in the tested limit concentration and the control after 24 h and 48 h. The immobilisation rates in % after 24 h and 48 h of exposure are given in the table below. There was no biologically significant effect neither in the tested limit concentration nor

in the control group.

 Percentage Number of Daphnia Incapable of Swimming after 24 and 48 hours of Exposure

Test item Nominal loading of the test item [mg/L]	IMMOBILISATION [%]
	24 hours					48 hours
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
100 (Saturated solution)	0	0	0	0	0	0	0	0	20	5
Control	0	0	0	0	0	0	0	0	0	0

 

According to OECD Guidance Document No 23 (2000) if an effect cannot be detected in a test with a saturated solution the result has to be reported as no toxic effects at saturation. Exposures are expressed in terms of the original concentration of the test item in water at the start of the mixing period (loading rate).

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

At the saturated solution with a nominal loading of 100 mg/L of the test item , no biologically significant effect was determined to Daphnia magna.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution (100 mg/L) as limit concentration of the test item was determined according to OECD 202 (1999, Draft) and EC Directive 92/69/EC Method C.2 (1992) .

(Saturated solution: The maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, acc. to OECD Series, No. 23 (2000)6).

The limit test was conducted under static conditions over 48 h. 

20 test organisms were exposed to the limit concentration and control.

A reference test was carried out with potassium dichromate to determine the toxicity of the reference item. The EC50-value of the

reference item of 2.07 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS, DIN Guideline 38412 L 30. Water quality parameters pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

At the saturated solution with a nominal loading of 100 mg/L of the test item , no biologically significant effect was determined to Daphnia magna.