Reaction mass of 4-[[4-(aminocarbonyl)phenyl]azo]-N-(2-ethoxyphenyl)-3-hydroxynaphthalene-2-carboxamide and 4-[[4-(aminocarbonyl)phenyl]azo]-3-hydroxy-N-(2-methoxyphenyl)naphthalene-2-carboxamide

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Remarks:

Pigment Red 022

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 guidelines and compliant to GLP, well documented translation of original report (in Japanese)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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 immediately prior to water renewal (48 hr), and analyzed by HPLC.

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 transferred to the test vessel by rinsing with dilution water and adjusted to 5 L.
The prepared test solution showed red color due to the test substance original 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):

Oryzias latipes

Details on test organisms:

Species: Japanese killifish
Scientific name: Oryzias latipes
Body length: 2.0 cm (1.7 - 2.1 cm), n = 10
Body Weight: 0.16 g (0.14 - 0.18 g), n = 10
Supplier: Izumoto fish firm (Osaka, Japan)
Date of receipt: December 18, 2000

Acclimation: December 18, 2000 - January 22, 2001
Mortality during 7 days before the initiation of exposure was less than 5%. Visibly healthy and normal fishes were used for the test.
Condition for acclimation
Culturing water: Dilution water
System: Recirculatory filtering system
Temperature: 24 +/- 1 deg C
Lighting: Indoor lighting, 16 hr light / 8 hr dark
Feeding: Commercially available feed (Trade name: TetraMin)
Feed amount: ca. 2% / body weight / day (until the final 24 hours before the initiation of exposure)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

Total hardness: 63.1 mg/L

Test temperature:

23.8 - 24.0 deg C

pH:

7.4 - 7.8

Dissolved oxygen:

In mg/L:
Control: 6.7 (old medium, 96 hours) to 8.1
Solvent control: 6.5 (old medium, 96 hours) to 8.1
Exposure group (100 mg/L): 5.4 (old medium, 96 hours) to 8.0

Nominal and measured concentrations:

Nominal limit concentration: 100 mg/L;
Measured concentration test start (new medium): 100 mg/L;
Measured concentration at 48 hours (old medium): 99 mg/L;
Geometric mean measured concentration: 100 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:

Test conditions
Type of exposure: Semi-static (water renewal was made after 48 hr exposure)
Duration of exposure: 96 hr
Volume of test solution: 5 L / vessel
Replication: 1 vessel / test group
Number of organisms: 10 fish / test group
Aeration: None
Temperature: 24 +/- 1 deg C
Lighting: Indoor lightning, 16-hour light / 8-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.

Test vessels and water bath
Vessel: 5 L volume whole glass aquarium (inside size approximately 21 x 16 x 23 cm)
Water bath: FRP Thermostatic water bath (inside size 300 x 81.5 x 28.5 cm, controlled with warm and cold water)
Dissolved oxygen analyzer: Model 58 (YSI Inc.)

Selection of test concentration
In a range-finding study conducted at the concentration of 10 and 100 mg/L, 0% mortalities were observed even at maximum exposure concentration of 100 mg/L, and no toxicological symptoms were observed. Based on these results, a limit test with 100 mg/L, which was the maximum concentration tested in the range-finding study, 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.

The body weight and body length of the 10 fishes, which were acclimated in the same aquarium used for culturing, were measured before the initiation of exposure in the study.
After the measurement of temperature, dissolved oxygen and pH of test water in each test vessel, ten fishes for each test group were introduced to each test vessel. The temperature, dissolved oxygen and pH were measured every day during the exposure period. In addition, the temperature, dissolved oxygen and pH were also measured before and after water renewal. Fish were not fed during the exposure period.
Mortality and toxic symptoms or abnormity were recorded at 24, 48, 72 and 96 hr after the initiation of the study. The following were the common symptoms and definitions:
Death: No reaction to the spurs was observed.
Abnormal breath: Different movement of gill cover is observed compared to control group.
Abnormal swimming: Different swimming form is observed compared to control group (e.g., slow-moving, hypersensitivity, convulsion and inversion etc.).
Inability to swim: Lack of swimming capability is observed although it is moving at surface or bottom of the water. State of overturn and apparent death are included.

Reference substance (positive control):

yes

Remarks:

copper (II) sulfate pentahydrate

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

EC0

Effect conc.:

100 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

behaviour

Remarks:

& mobility

Details on results:

The measured concentrations of the test substance in the test solution were 100% and 99% of the nominal at the initiation of exposure and 48 hr after the test initiation, respectively.
Since 0% mortality in the control groups was observed during the study, the conditions for the validity of the study were fulfilled.
During the exposure period, toxic symptoms were not observed at 100 mg/L test item concentration. Also for the controls, no abnormal symptoms were observed.
Water temperature during exposure period was between 23.8 and 24.0 deg C, consequently, the temperature of all groups was within a range of pre-set temperature, 24 +/- 1 deg C. Dissolved oxygen level was in the range from 5.4 to 8.1 mg/L and more than 60% of the saturation concentration (8.25 mg/L at 24.0 deg C) were maintained in all groups. The variation of pH values was slight and in the range from 7.3 to 7.8 during the exposure.

Results with reference substance (positive control):

The median lethal concentration (LC50) at 96 hr with a reference substance (copper (II) sulfate pentahydrate, reagent grate, Lot No. PAH2068, Wako Pure Chemical Industries) was 2.3 mg/L, which was comparable to the historical LC50 of 2.3 -5.7 mg/L, (n = 8) obtained at the testing facility since December, 1995.

Reported statistics and error estimates:

No statistical analysis was applied for the calculation of median lethal concentration (LC50), since the study was conducted as a limit test at 100 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

In a reliable 96-hours acute toxicity study on Oryzias latipes performed according to OECD 203 and compliant to GLP (reliability category 1), the test item caused neither lethality nor abnormal behaviour or mobility at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 100 mg/L). Also no lethality nor abnormal behaviour or mobility was observed 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):
LC50 (96 h): > 100 mg/L;
LC0 (96 h): 100 mg/L;
EC0 (96 h, behaviour & mobility): 100 mg/L.

Executive summary:

In a reliable 96-hours acute toxicity study on Oryzias latipes with semi-static test design (medium renewal after 48 hours) performed according to OECD 203 and compliant to GLP (reliability category 1), the test item caused neither lethality nor abnormal behaviour or mobility at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 100 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. Also no lethality nor abnormal behaviour or mobility was observed 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):

LC50 (96 h): > 100 mg/L;

LC0 (96 h): 100 mg/L;

EC0 (96 h, behaviour & mobility): 100 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 2

Endpoint:

short-term toxicity to fish

Remarks:

Pigment Red 022

Type of information:

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

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 guidelines and compliant to GLP, well documented translation of original report (in Japanese)

Justification for type of information:

See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 210 (Chapter 13)

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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 immediately prior to water renewal (48 hr), and analyzed by HPLC.

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 transferred to the test vessel by rinsing with dilution water and adjusted to 5 L.
The prepared test solution showed red color due to the test substance original 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):

Oryzias latipes

Details on test organisms:

Species: Japanese killifish
Scientific name: Oryzias latipes
Body length: 2.0 cm (1.7 - 2.1 cm), n = 10
Body Weight: 0.16 g (0.14 - 0.18 g), n = 10
Supplier: Izumoto fish firm (Osaka, Japan)
Date of receipt: December 18, 2000

Acclimation: December 18, 2000 - January 22, 2001
Mortality during 7 days before the initiation of exposure was less than 5%. Visibly healthy and normal fishes were used for the test.
Condition for acclimation
Culturing water: Dilution water
System: Recirculatory filtering system
Temperature: 24 +/- 1 deg C
Lighting: Indoor lighting, 16 hr light / 8 hr dark
Feeding: Commercially available feed (Trade name: TetraMin)
Feed amount: ca. 2% / body weight / day (until the final 24 hours before the initiation of exposure)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

Total hardness: 63.1 mg/L

Test temperature:

23.8 - 24.0 deg C

pH:

7.4 - 7.8

Dissolved oxygen:

In mg/L:
Control: 6.7 (old medium, 96 hours) to 8.1
Solvent control: 6.5 (old medium, 96 hours) to 8.1
Exposure group (100 mg/L): 5.4 (old medium, 96 hours) to 8.0

Nominal and measured concentrations:

Nominal limit concentration: 100 mg/L;
Measured concentration test start (new medium): 100 mg/L;
Measured concentration at 48 hours (old medium): 99 mg/L;
Geometric mean measured concentration: 100 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:

Test conditions
Type of exposure: Semi-static (water renewal was made after 48 hr exposure)
Duration of exposure: 96 hr
Volume of test solution: 5 L / vessel
Replication: 1 vessel / test group
Number of organisms: 10 fish / test group
Aeration: None
Temperature: 24 +/- 1 deg C
Lighting: Indoor lightning, 16-hour light / 8-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.

Test vessels and water bath
Vessel: 5 L volume whole glass aquarium (inside size approximately 21 x 16 x 23 cm)
Water bath: FRP Thermostatic water bath (inside size 300 x 81.5 x 28.5 cm, controlled with warm and cold water)
Dissolved oxygen analyzer: Model 58 (YSI Inc.)

Selection of test concentration
In a range-finding study conducted at the concentration of 10 and 100 mg/L, 0% mortalities were observed even at maximum exposure concentration of 100 mg/L, and no toxicological symptoms were observed. Based on these results, a limit test with 100 mg/L, which was the maximum concentration tested in the range-finding study, 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.

The body weight and body length of the 10 fishes, which were acclimated in the same aquarium used for culturing, were measured before the initiation of exposure in the study.
After the measurement of temperature, dissolved oxygen and pH of test water in each test vessel, ten fishes for each test group were introduced to each test vessel. The temperature, dissolved oxygen and pH were measured every day during the exposure period. In addition, the temperature, dissolved oxygen and pH were also measured before and after water renewal. Fish were not fed during the exposure period.
Mortality and toxic symptoms or abnormity were recorded at 24, 48, 72 and 96 hr after the initiation of the study. The following were the common symptoms and definitions:
Death: No reaction to the spurs was observed.
Abnormal breath: Different movement of gill cover is observed compared to control group.
Abnormal swimming: Different swimming form is observed compared to control group (e.g., slow-moving, hypersensitivity, convulsion and inversion etc.).
Inability to swim: Lack of swimming capability is observed although it is moving at surface or bottom of the water. State of overturn and apparent death are included.

Reference substance (positive control):

yes

Remarks:

copper (II) sulfate pentahydrate

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

EC0

Effect conc.:

100 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

behaviour

Remarks:

& mobility

Details on results:

The measured concentrations of the test substance in the test solution were 100% and 99% of the nominal at the initiation of exposure and 48 hr after the test initiation, respectively.
Since 0% mortality in the control groups was observed during the study, the conditions for the validity of the study were fulfilled.
During the exposure period, toxic symptoms were not observed at 100 mg/L test item concentration. Also for the controls, no abnormal symptoms were observed.
Water temperature during exposure period was between 23.8 and 24.0 deg C, consequently, the temperature of all groups was within a range of pre-set temperature, 24 +/- 1 deg C. Dissolved oxygen level was in the range from 5.4 to 8.1 mg/L and more than 60% of the saturation concentration (8.25 mg/L at 24.0 deg C) were maintained in all groups. The variation of pH values was slight and in the range from 7.3 to 7.8 during the exposure.

Results with reference substance (positive control):

The median lethal concentration (LC50) at 96 hr with a reference substance (copper (II) sulfate pentahydrate, reagent grate, Lot No. PAH2068, Wako Pure Chemical Industries) was 2.3 mg/L, which was comparable to the historical LC50 of 2.3 -5.7 mg/L, (n = 8) obtained at the testing facility since December, 1995.

Reported statistics and error estimates:

No statistical analysis was applied for the calculation of median lethal concentration (LC50), since the study was conducted as a limit test at 100 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

In a reliable 96-hours acute toxicity study on Oryzias latipes performed according to OECD 203 and compliant to GLP (reliability category 1), the test item caused neither lethality nor abnormal behaviour or mobility at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 100 mg/L). Also no lethality nor abnormal behaviour or mobility was observed 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):
LC50 (96 h): > 100 mg/L;
LC0 (96 h): 100 mg/L;
EC0 (96 h, behaviour & mobility): 100 mg/L.

Executive summary:

In a reliable 96-hours acute toxicity study on Oryzias latipes with semi-static test design (medium renewal after 48 hours) performed according to OECD 203 and compliant to GLP (reliability category 1), the test item caused neither lethality nor abnormal behaviour or mobility at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 100 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. Also no lethality nor abnormal behaviour or mobility was observed 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):

LC50 (96 h): > 100 mg/L;

LC0 (96 h): 100 mg/L;

EC0 (96 h, behaviour & mobility): 100 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).

Description of key information

PR022

In a reliable 96-hours acute toxicity study on Oryzias latipes performed according to OECD 203 and compliant to GLP (reliability category 1), the test item caused neither lethality nor abnormal behaviour or mobility at the applied limit concentration of nominal 100 mg/L (geometric mean measured concentration: 100 mg/L). Also no lethality nor abnormal behaviour or mobility was observed 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):
LC50 (96 h): > 100 mg/L;
LC0 (96 h): 100 mg/L;
EC0 (96 h, behaviour & mobility): 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 100 mg/L