N-(5-chloro-2-methoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-[(phenylamino)carbonyl]phenyl]azo]naphthalene-2-carboxamide

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 09 NOV 2004 to 11 NOV 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study compliant to GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1999, Draft

Deviations:

yes

Remarks:

No test item analysis due to very poor water solubility of test item

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

No test item analysis due to very poor water solubility of test item

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP according to German Chemikaliengesetzt and Directive 88/320/EEC

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Stock solution: The solution (100 mg/L test item was weighed out) was prepared with dilution water one day prior to application.
Dispersion treatment: The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 µm (RC, SCHLEICHER & SCHUELL).
Limit concentration: Saturated solution (100 mg/L)
Due to the very poor water solubility of the test item, the filtrated saturated solution prepared at nominal 100 mg/L was used in the limit test.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test organism: Daphnia magna STRAUS (Clone 5)
Reason for the selection: Daphnia magna STRAUS belongs, in accordance with the study of the test organism guideline, to the suitable Daphnia types and are bred in the test facility.
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder: DR.U.NOACK-LABORATORIEN, 31157 Sarstedt, Germany
Culture: In 2-3 L glass vessels with approximatively 1.8 L culture medium, at 21 deg C (temperatures of 20 - 25 deg C are tolerated), in an incubator, 16 h illumination, illumination strength max. 20 µmol photons*mE(-2)*sE(-1).
Culture medium: Culture medium Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
Feeding: At least 5x weekly ad libitum with a mix of Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 10E(6) cells/mL.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

no

Hardness:

249 mg CaCO3/L

Test temperature:

20.5 deg C (18-22 deg C, +/- 1 deg C)

pH:

Treatment
Start: 7.72 End: 7.59 - 7.61
Control
Start: 7.85 End: 7.60 - 7.65

Dissolved oxygen:

[mg/L]
Treatment
Start: 8.49 End: 7.98 - 8.20
Control
Start: 8.66 End: 7.87 - 8.06

Nominal and measured concentrations:

Due to the very poor solubility of the test item no test item analysis was carried out.

Details on test conditions:

Test vessels: Glass beakers (5 cm ID x 8 cm H), 50 mL volume were used.
Volume of the study medium: 20 mL

Dilution water: According to EC 92/69 L383A C.2. Annex
Component - Concentration [mg/L] in dilution water
KCL 5.76
NaHCO3 64.8
CaCl2*2 H2O 294
MgSO4*7 H2O 123

Number of study organisms per concentration and control: 20 animals, divided into 4 (2*) parallel samples, each with 5 (10*) animals
Number of animals per test vessel: 5 (10*)
Number of parallel samples per concentration: 4 (2*)
Age of test organisms: 2 - 24 h old
Acclimation: Ca. 2 h
Test temperature: 18 - 22 deg C, +/- 1 deg C constant
Illumination: Diffuse light, illumination range max. 20 µE*mE(-2)*sE(-1)
Photoperiod: 16/8 h light/dark cycle
Feeding: The daphnids were not fed during the study.

The study was performed as a limit test with saturated solution (100 mg/L)# under static conditions to enable the determination of immobilisation after 24 and 48 hours.
2 to 24 h old daphnids were used for the definitive study. They were obtained by removing the mother animals twice within 22 h.
The daphnids were acclimated to the dilution water ca. 2 hours before test start. The daphnids were not fed during the study.
Control replicates with dilution water (test medium without test item) were tested under the same conditions as the test groups.
20 daphnids divided into 4 (2*) groups with 5 (10*) animals per test vessel were used. The immobilisation of the daphnids was observed after 24 and 48 hours of exposure. Water parameter were measured at begin and end of the study.
The reference item was tested in five concentrations within the concentration range 0.58 to 5.8 mg/L.

*) The information in brackets refers to the test of the reference item
# 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.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate p.a. (Merck) 100% purity, Batch No. K 28974764

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Neither for the controls nor for the treatment replicates were any immobilzed daphnids observed.

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.

Reported statistics and error estimates:

EC10- and EC-50-values were only calculated for the reference analysis item via probit according to WEBER (1986).
Calculation of the confidence intervals were carried out using standard procedures according to BREITIG & TÜMPLING (1982).

BREITIG G. & TÜMPLING W. (1982): Methoden der Wasseruntersuchungen Bd.II, VEB Gustav Fischer Verlag, Jena.
WEBER E. (1986): Grundriss der biologischen Statistik, 9. Auflage

Validity criteria fulfilled:

yes

Conclusions:

In an acute immobilisation test to Daphnia magna according to OECD 202 (1999, draft) no effects on daphnids were observed at the nominal limit concentration of 100 mg/L. Due to the very poor water solubility of the submission substance, a filtrated saturated solution prepared at 100 mg/L was used.
EC50 (48h) > 100 mg/L nominal concentration
NOEC (48h) = 100 mg/L nominal concentration

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 submission substance, was determined compliant to GLP according to OECD 202 (1999, draft) and EC Directive 92/69/EC Method C.2 (1992) (reliability category 1). The limit test was conducted under static conditions over 48 h. 20 test organisms were exposed to the limit concentration and control. No test item analysis was carried out. 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 (100 mg/L) of the test item no biologically significant effect was determined. Neither for the control nor for the treatment replicates any immobilized daphnids could be observed.

#) 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.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 22 MAR 2005 to 24 MAR 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study compliant to GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

No test item analysis due to very poor water solubility of test item

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

No test item analysis due to very poor water solubility of test item

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP according to German Chemikaliengesetzt and Directive 88/320/EEC

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Stock solution: The solution (100 mg/L test item was weighed out) was prepared with dilution water one day prior to application.
Dispersion treatment: The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 µm (RC, SCHLEICHER & SCHUELL).
Limit concentration: Saturated solution (100 mg/L)
Due to the very poor water solubility of the test item, the filtrated saturated solution prepared at nominal 100 mg/L was used in the limit test.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test organism: Daphnia magna STRAUS (Clone 5)
Reason for the selection: Daphnia magna STRAUS belongs, in accordance with the study of the test organism guideline, to the suitable Daphnia types and are bred in the test facility.
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder: DR.U.NOACK-LABORATORIEN, 31157 Sarstedt, Germany
Culture: In 2-3 L glass vessels with approximatively 1.8 L culture medium, at 21 deg C (temperatures of 20 - 25 deg C are tolerated), in an incubator, 16 h illumination, illumination strength max. 20 µE*mE(-2)*sE(-1).
Culture medium: Culture medium Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
Feeding: At least 5x weekly ad libitum with a mix of Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 10E(6) cells/mL.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

no

Hardness:

266 mg CaCO3/L

Test temperature:

20.8 deg C (18-22 deg C, +/- 1 deg C)

pH:

Treatment
Start: 7.72 End: 7.60 - 7.69
Control
Start: 7.8 End: 7.83 - 7.94

Dissolved oxygen:

[mg/L]
Treatment
Start: 8.9 End: 8.09 - 8.14
Control
Start: 8.69 End: 7.67 - 7.93

Nominal and measured concentrations:

Due to the very poor solubility of the test item no test item analysis was carried out.

Details on test conditions:

Test vessels: Glass beakers (5 cm ID x 8 cm H), 50 mL volume will be used.
Volume of the study medium: 20 mL

Dilution water: According to EC 92/69 L383A C.2. Annex
Component - Concentration [mg/L] in dilution water
KCL 5.76
NaHCO3 64.8
CaCl2*2 H2O 294
MgSO4*7 H2O 123

Number of study organisms per concentration and control: 20 animals, divided into 4 (2*) parallel samples, each with 5 (10*) animals
Number of animals per test vessel: 5 (10*)
Number of parallel samples per concentration: 4 (2*)
Age of test organisms: 2 - 24 h old
Acclimation: Ca. 2 h
Test temperature: 18 - 22 deg C, +/- 1 deg C constant
Illumination: Diffuse light, illumination range max. 20 µE*mE(-2)*sE(-1)
Photoperiod: 16/8 h light/dark cycle
Feeding: The daphnids were not fed during the study.

A static preliminary range finding test was conducted as limit test with the saturated solution (NON-GLP-state). The definitive study was performed as a limit test with saturated solution (100 mg/L) under static conditions to enable the determination of immobilisation after 24 and 48 hours.
2 to 24 h old daphnids were used for the definitive study. They were obtained by removing the mother animals twice within 22 h.
The daphnids were acclimated to the dilution water ca. 2 hours before test start. The daphnids were not fed during the study.
Control replicates with dilution water (test medium without test item) were tested under the same conditions as the test groups.
20 daphnids divided into 4 (2*) groups with 5 (10*) animals per test vessel will be used. In the definitive study the immobilisation of the daphnids was observed after 24 and 48 hours of exposure. Water parameter were measured at begin and end of the study.
The reference item was tested in five concentrations within the concentration range 0.58 to 5.8 mg/L.

*) The information in brackets refers to the test of the reference item

Reference substance (positive control):

yes

Remarks:

Potassium dichromate p.a. (Merck) 100% purity, Batch No. K 28974764

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Neither for the controls nor for the treatment replicates were any immobilzed daphnids observed.

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 : 1.51
EC50 : 1.84 (CI 1.74 - 1.95)
EC100 : 3.20
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.

Reported statistics and error estimates:

EC10- and EC-50-values were only calculated for the reference analysis item via probit according to WEBER (1986).
Calculation of the confidence intervals were carried out using standard procedures according to BREITIG & TÜMPLING (1982).

BREITIG G. & TÜMPLING W. (1982): Methoden der Wasseruntersuchungen Bd.II, VEB Gustav Fischer Verlag, Jena.
WEBER E. (1986): Grundriss der biologischen Statistik, 9. Auflage

Validity criteria fulfilled:

yes

Conclusions:

In an acute immobilisation test to Daphnia magna according to OECD 202 no effects on daphnids were observed at the nominal limit concentration of 100 mg/L. Due to the very poor water solubility of the submission substance, a filtrated saturated solution prepared at 100 mg/L was used.
EC50 (48h) > 100 mg/L nominal concentration
NOEC (48h) = 100 mg/L nominal concentration

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 submission substance, was determined compliant to GLP according to OECD 202 (2004) and EC Directive 92/69/EC Method C.2 (1992) (reliability category 1). The limit test was conducted under static conditions over 48 h. 20 test organisms were exposed to the limit concentration and control. No test item analysis was carried out. 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 1.84 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 (100 mg/L) of the test item no biologically significant effect was determined. Neither for the control nor for the treatament replicates any immobilized daphnids could be observed.

#) 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.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2000-12-13 to 2001-04-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to relevant guideline and compliant to GLP, well documented translation of original report (in Japanese)

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Remarks:

The study was conducted in accordance with "Eco-toxicity test Standards" by Japanese Ministry of the Environment.

Analytical monitoring:

yes

Details on sampling:

Samplings were made at the time point of study initiation (0 hr) and termination (48 hr).

Vehicle:

yes

Details on test solutions:

After weighing of a necessary amount of test substance using an electronic balance, the same amount of hydrogenated castor oil (HCO-40) was added to the test substance as co-solvent and mixed well. Then, the solution was made up to the volume with dilution water and mixed with a stirrer for 30 minutes to prepare the test solution of the desired nominal concentration.
The prepared test solutions showed strong red color but no visible precipitate was observed. The solution of control and solvent control was observed to be clear and colorless without visible undissolved test substance.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Water fleas
Scientific name: Daphnia magma
Supplier: National Institute for Environmental Studies, Japan
Date of receipt: October 28, 1996
Maintenance after received: Subculture
Acclimation: Acclimation period: December 28, 2000 - January 16, 2001
Mortality of parent daphnids during acclimation period was 0% and ephippia and presence of males were not observed.
Age: Juvenile within 24-hr of age

Condition for acclimation
Water: Dilution water
Loading: 100 daphnids per 5 L culturing water
Temperature: 20 +/- 1 deg C
Lighting: Indoor lighting, 16 hr light 18 hr dark
Feeding: Chlorella vulgaris (commercially available condensed chlorella liquid, trade name: raw chlorella-V12, manufacture: Chlorella Industry Co., Ltd.). The amount of organic carbon in 1% suspension was 1192 mg C/L (measured on December, 2000). The amount fed was determined based on this value. The measurement of the amount of organic carbon was contracted out to Koei Techno Company, Limited.
Feed amount: 0.1 - 0.2 mg C (total organic carbon) per daphnid per day.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Total hardness 63.1 mg/L

Test temperature:

Temperature: 20 +/- 1 deg C

pH:

8.0 to 8.1

Dissolved oxygen:

8.6 to 8.9 mg/L

Nominal and measured concentrations:

Nominal limit concentration: 100 mg/L;
Measured concentration test start: 100 mg/L;
Measured concentration test end (48 hours): 120 mg/L;
Geometric mean measured concentration: 110 mg/L.
The concentration of test substance in the control and solvent control were less than the quantification limit (0.06 mg/L).

Details on test conditions:

- Volume of test solution: 100 mL / vessel (glass beaker)
- Replication: 4 vessel / test group
- Number of organisms: 20 daphnids per test group (5 daphnids per vessel)
- Lighting: Indoor lightning, 16-hour light, 18-hour dark
- Feeding: None
- Dilution water: Tap water (Takarazuka city, Hyogo) was dechlorinated with charcoal filter and saturated with oxygen by aeration. Then the water was used as the dilution water after adjusting to the test temperature. The results of water quality measurement are shown in section "Any other information on materials and methods incl. tables". From the results of the water quality measurement and no observed abnormalities in organisms, it was judged that the dilution water can be used as culturing water in the bio-assays.
- Water bath: Thermostatic water bath for daphnid test
- Selection of test concentration: In a range-finding study conducted at the concentration of 10 mg/L and 100 mg/L (which was the maximum attainable homogeneous dispersion concentration), no immobilizations were observed among daphnids exposed to all the exposure levels. Based on these results, a limit test with 100 mg/L was chosen for the definitive study. As reference, a control group (dilution water only) and a solvent control group, in which co-solvent concentration was 100 mg/L (i.e. the same as for the exposure group) were allocated in the study.
- Test operation procedure: The prepared test solutions were dispensed to 4 vessels with 100 mL per each test concentration, and the vessels were placed in the thermostatic water bath. The initiation of exposure was defined as the time point at which daphnids were introduced to each vessel using glass pipette after measurement of temperature, dissolved oxygen level and pH of test solution. The pipetted aliquot of water was less than approximately 1% of the volume of test solution, which was used as guide. The observation of number of immobilized daphnids was made at 24 and 48 hours after the initiation of exposure. The daphnid was judged as immobilized when the daphnid was not able to swim within 15 seconds after gentle agitation of the test vessel (The swimming was defined that the daphnid was able to swim in water. Therefore, the daphnid crawling and moving along the bottom of the vessel was judged as immobilized in the study. As for the floating of daphnid on the surface, it was judged as immobilized in case that although the daphnid was able to swim after immersed in the water forcibly by water drops, came up to water surface again. And, it was not
judged as immobilized in case that the daphnid swam in water at least once in 15 seconds, even if its swim was not normal).
The temperature, dissolved oxygen and pH of test solution of all test groups were measured at the initiation and termination of exposure.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 110 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

110 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 110 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Since the immobilization rate and the rate of floating daphnid on the surface in both the control and solvent control group at 48 hours exposure were 0%, it was judged that the study was conducted under appropriate conditions.
No immobilization occurred in the treatment replicates.

Results with reference substance (positive control):

The median effective concentration (EC50) at 48 hr with the reference substance potassium dichromate (reagent grade, Lot No. ACQ2610, Wako Pure Chemical Industries) was 0.64 mg/L, which was comparable to the historical EC50 of 0.38 - 0.94 mg/L, n = 10) obtained at the testing facility since December, 1995.

Reported statistics and error estimates:

No statistical analysis was applied for the calculation of median effective concentration (EC50), since the study was conducted as a limit test at 100 mg/L.
The no-observed-effect concentration (NOEC) was defined as the highest concentration where no adverse effect on daphnids was observed, and the lowest
concentration producing 100% immobilization was defined as the lowest concentration (EC100) where all daphnids were observed immobilized.

Validity criteria fulfilled:

yes

Conclusions:

In a reliable 48-hours acute toxicity study on Daphnia magna performed according to OECD 202 and compliant to GLP (reliability category 1), the test item caused no immobilization at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 110 mg/L). No immobilization occured in the control and the solvent control (containing the amount of HCO-40 used in the treatment group). The following (no)effect concentrations were determined (geometric mean measured concentrations):
EC50 (48 h): > 110 mg/L
LOEC (48 h): > 110 mg/L
NOEC (48 h): 110 mg/L

Executive summary:

In a reliable 48-hours acute toxicity study on Daphnia magna performed according to OECD 202 and compliant to GLP (reliability category 1), the test item caused no immobilization at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 110 mg/L). Hydrogenated castor oil (HCO-40) was used at equal weight as co-solvent to achieve the limit concentration of the otherwise insoluble test item. No immobilization occured in the control and the solvent control (containing the amount of HCO-40 used in the treatment group). The following (no)effect concentrations were determined (geometric mean measured concentrations):

EC50 (48 h): > 110 mg/L

LOEC (48 h): > 110 mg/L

NOEC (48 h): 110 mg/L

The robust study summary is based on the translation of the original study report performed by the same company involved with performance of the test (Sumika TechnoService Corporation, Japan).

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Refer to the analogue approach provided in the CSR.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 110 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other:

Remarks:

Sumika, 2001

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other:

Remarks:

Noack, 2004

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other:

Remarks:

Noack, 2005

Description of key information

EC50 for freshwater invertebrates: > water solubility limit

Additional information

Three reliable (reliability category 1) studies on the acute toxicity towards invertebrates (according to OECD 202, compliant to GLP) with three different pigments (Pigment Red 2, 22 and 146) are available, meeting the validity criteria of the OECD guideline. Static test design was applied. Pigments Red 2 and 146 were tested as saturated solutions at the nominal limit concentration of 100 mg/L. Pigment Red 22 was tested as pigment dispersion at the geometric mean measured concentration of 110 mg/L (100 mg/L nominal concentration; hydrogenated castor oil (HCO-40) was used at equal weight as co-solvent to achieve the limit concentration of the otherwise insoluble test item). No toxicity to Daphnia magna could be found consistently in these tests for any of these pigments. Therefore, the Naphtol-AS Pigments should be regarded as acutely nontoxic to invertebrates.