Reaction products of diazotized 2-amino-4-(methylsulfonyl)-phenol with 2-(3-Chlorophenyl)-2,4-dihydro-5-methyl-3H-pyrazol-3-one and Methyl-7-hydroxy-1-naphthylcarbamate – and chromium (III) 2:1, salted by ammonium, sodium and acid

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study including analytical monitoring

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of diazotized 2-amino-4-(methylsulfonyl)-phenol with 2-(3-Chlorophenyl)-2,4-dihydro-5-methyl-3H-pyrazol-3-one and Methyl-7-hydroxy-1-naphthylcarbamate – and chromium (III) 2:1, salted by ammonium, sodium and acid

EC Number:

936-609-1

Molecular formula:

C40H32CrN4O12S2-R+] . [C36H28ClCrN7O10S2-R+] . [C34H26Cl2CrN8O8S2-R+]; R+ = NH4+, Na+ or H+

IUPAC Name:

Reaction products of diazotized 2-amino-4-(methylsulfonyl)-phenol with 2-(3-Chlorophenyl)-2,4-dihydro-5-methyl-3H-pyrazol-3-one and Methyl-7-hydroxy-1-naphthylcarbamate – and chromium (III) 2:1, salted by ammonium, sodium and acid

Details on test material:

- Substance type:
- Physical state: Brown powder
- Stability under test conditions: Not specified
- Storage condition of test material: Room temperature, protected from light, in original container

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

All test concentrations and the control were analytically verified by HPLC-DAD in the fresh (at 0 hours) and old media (after 48 hours) as specified below.
Sampling from freshly prepared media was carried out immediately after preparation of the test concentrations.
Sampies of the old media were taken directly from additional replicates prepared with test media, but without daphnids, incubated under test conditions until sampling, because volumes of the test vessels were not sufficient for the concentration-control analysis.
All original samples were stored at 6 ± 2 °C until sample preparation, if necessary. Prepared samples were stored in the auto sampler until analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
A stock solution (10.0 mg/L of the test item were weighed out) was freshly prepared with dilution water.
The stock solution was stirred for 2 hours at 1100 rpm with a magnetic stirrer at room temperature. Thereafter, the stock solution was centrifuged for 10 min at 3000 rpm to separate undissolved test item. The suitability of this procedure was confirmed by a matching Chromatographie profile between an analytical standard and a test item solution. This fact indicates that there is no shift in the chemical composition between the complete dissolved test item in the calibration standards and the test item solution obtained by the described dispersion treatment.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Daphnia magna STRAUS (Clone 5)
- 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 were used. They were obtained by removing the parental daphnids twice within 22 h.
- Method of breeding: ln 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/m^2*s, pH in the range of 6 to 9.
- Culture medium: Elendt M4, according to ELENDT ( 1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
- Culture 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 > 10^6 cells/ml.
- Feeding during test: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

160 to 180 mg CaCO3/L

Test temperature:

18 - 22 °C, ± 1 °C constant

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 ml capacity
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Same as for culture.
- Intervals of water quality measurement: At the start of the exposure (0 h), the water quality parameters (pH value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured.
At the beginning of the exposure period (0 h), the water quality parameters of the fresh media (pH value, oxygen concentration) were measured in one additional replicate per concentration Ievei and control.
After 48 h, the water parameters of the old media were measured in all replicates per concentration Ievei and control.

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h Iight/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 E/m^2 * s

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
After 48 h, the water parameters of the old media were measured in all replicates per concentration Ievei and control.
lmmobilization was determined in all.groups after 24 and 48 h.
An animal was considered to be immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel.

TEST CONCENTRATIONS
5 test item concentrations, prepared by diluting the stock solution of 10.0 mg/L with dilution water, were tested as follows:
0.100 - 0.320- 1.00 - 3.20- 10.0 mg/L (dilution factor √10).
The test concentrations were based on the results of a non GLP preliminary range finding test conducted at the test facility under static conditions.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.339 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

0.113 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

1.66 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

All test item solutions were visually clear throughout exposure. The concentration Ieveis of 0.320 to 10.0 mg/L showed a concentration related increase in the brownish colour. The concentration Ievei of 0.100 mg/L was colourless.

Results with reference substance (positive control):

The EC50-value for the most recent of the monthly performed reference tests was:
EC50 : 1.74 mg/L (CI 1.49- 2.00 mg/L)
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 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.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test substance is acutely very toxic to aquatic invertebrates.