Reaction mass of sodium [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) and sodium bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) and sodium bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(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:

2016-09-21 to 2017-05-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004-11-23

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

No 440/2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of sodium [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) and sodium bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) and sodium bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(1-)

EC Number:

915-758-6

Molecular formula:

C32H18CrN6O8.Na / C32H22CrN10O8.Na / C32H20CrN8O8.Na

IUPAC Name:

Reaction mass of sodium [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) and sodium bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) and sodium bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(1-)

Test material form:

solid

Details on test material:

Appearance:
- physical state: solid
- color: dark brown

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

Sampling and analysis

Analytical monitoring:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Strain/clone: Daphnia magna STRAUS
- Source: The clone of Daphnia magna STRAUS 1820 used was supplied by the Institut National de Recherche Chimique Appliquee, France, in 1978. From this date on this clone was cultured and bred continuously in the Ecotoxicology Laboratory of Experimental Toxicology and Ecology, BASF SE, Ludwigshafen Germany.
- Age of parental stock (mean and range, SD): approx. 20 - 30 parthenogenetically reproducing females for a maximum of 4 weeks.
- Age at start of exposure: < 24 hours (at least 3rd brood progeny)
- Feeding during test: No feeding during the exposure period.

ACCLIMATION
- Acclimation conditions (same as test or not): The Daphnia are cultured under the identical conditions as the test including test media (Elendt M4), water quality, and temperature (20 ± 1 °C).

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
All individuals in each mass culture originate from a single female and are thus genetic clones. After approximately 14 days the adults have produced at least 3 broods and the young can be used in tests. Offspring are removed from the mass cultures at least once daily during the normal work week to ensure that young daphnia are <24-h old (first instar) at start of the exposure.

Study design

Test type:

semi-static

Water media type:

freshwater

Remarks:

A synthetic fresh water (Elendt M4) is used as media for culture and test purposes. For the composition of this M4 medium see OECD 202.

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

2.20 - 3.20 mmol/L

Test temperature:

19.7 - 20.1 °C

pH:

7.5 - 8.5

Dissolved oxygen:

8.7 - 9.4 mg/L

Conductivity:

550 - 650 µS/cm

Nominal and measured concentrations:

100 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: Numbered glass beakers (nominal volume 100 mL)
- Fill volume: 50 mL
- Aeration: ~ 24 hours prior to use.
- Renewal rate of test solution (frequency): 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 0.1 animals/mL

TEST MEDIUM / WATER PARAMETERS
- Total organic carbon: <2 mg/L
- Ca/Mg ratio: about 4:1
- Conductivity: 550 - 650 µS/cm
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light : 8 hours darkness
- Light intensity: 161 - 677 lux at a wave length of 400 - 750 nm

EFFECT PARAMETERS MEASURED: Mobility after 0, 24, and 48 h

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: In a preliminary test (experimental conduct in accordance with GLP, without a GLP status, a concentration of 100 mg/L caused no immobilisation or symptoms after 48 hours.
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Sodium chloride

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate

Basis for effect:

mobility

Details on results:

VALIDITY CRITERIA
This test was fully compliant with all the following validity criteria required by the corresponding test guidelines and is considered valid.
• ≤10% immobilisation in the control
• O2 concentration ≥3 mg/L in control and test vessels

Any other information on results incl. tables

 

Table 1: Number of mobile Daphnids

Nominal concentration [mg/L]	Number of mobileDaphniaat each observation period
	Replicate 1 [h]			Replicate 2 [h]			Replicate 3 [h]			Replicate 4 [h]			∑	∑
	0	24	48	0	24	48	0	24	48	0	24	48	24 h	48 h
0 (control)	5	5	5	5	5	5	5	5	5	5	5	5	20	20
100	5	5	5	5	5	4	5	5	5	5	5	5	20	19
Number of test animals per replicate: 5 Number of test animals per test group: 20

 

Table 2: Total organic carbon (TOC) content

Loading rate [mg/L]	after 0 h [mg/L]	after 24 h [mg/L]
0 (control)	0.9	1.1
100	3	3.1

The TOC in the treatment group was higher than in the control test medium. The higher TOC is likely due to soluble organic components from the test substance. The TOC value confirms qualitatively that the daphnids in the WAF were exposed to soluble organic components from the test substance.

Table 3: DIissolution Behavior

Test Group [mg/L]	0 hour	24 hours		48 hours
		Old test solution	New test solution
0 (control)	0	0	0	0
100	1	1	1	1

Explanation of abbreviations:

0: no remarkable observations, clear test medium

1: coloration caused by the test substance (yellow)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test item is with high probability not acutely harmful to aquatic invertebrates.