Anthraquinone

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29.9. - 13.10. 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP, analytical monitoring

Qualifier:

according to guideline

Guideline:

other: "Protocol for Conducting a Flow Through Acute Toxicity Test with Bluegill Sunfish following TSCA Guidelines", Protocol 011988/BG.FA.

Deviations:

yes

Remarks:

dissolved oxygen; feding of test organism, diluter malfunction occurred on test day 10 which resulted in no test material being delivered for approximately 21 hours.Since no mortalities were observed among organisms exposed for 14 days to the highest trea

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

The high, middle and low test concentrations and the control solution were sampled and analyzed for Anthraquinone concentrations prior to the start of the definitive exposure. Results of these pretest analyses were used to judge whether sufficient quantities of test material were being delivered and maintained in the exposure aquaria to initiate the definitive test. During the in life phase of the definitive study, water samples were removed from both replicate test solutions of each treatment level and the controls on test days 0 and 4 for analysis of Anthraquinone. Subsequently, on days 7 and 14, water samples were removed from both replicate test solutions of the two highest test concentrations and the control solutions. Each exposure solution sample was collected form the approximate midpoint of the aquarium with a volumetric pipet. In addition, quality assurance (QA) blind samples were prepared at each sampling interval and remained with the set of exposure solution samples through the analytical process. The QA samples were prepared in dilution water at Anthraquinone concentrations unknown to the analyst. Results of these analysis were used to judge the precision and quality control maintained during the analysis of exposure solution samples.

Vehicle:

yes

Details on test solutions:

A diluter stock solution of 1.34 mg A.I./mL was prepared by diluting 0.3358 g of Anthraquinone (0.3352 grams of active
ingredient) with acetone to a total volume of 250 mL. A Sage syringe pump in conjunction with a 50-mL gas-tight syringe was used to deliver 0.029 mL/min of the stock solution of Anthraquinone (1.34 mg A.I./mL) to the diluter's chemical mixing chamber receiving approximately 0.389 L/min dilution water. The mixing chamber was positioned over a magnetic stirrer, with an ultrasonic water bath. The continuous stirring and sonication
aided in the solubilization of the test material. The solution contained in the mixing chamber constituted the highest nominal test concentration (100 µg A.I./L) and was subsequently diluted (65% dilution factor) to provide the desired exposure concentration range. A similar system used to deliver the test material was also used to deliver acetone to the solvent control aquaria.
The solvent control solutions contained the maximum amount of acetone (CAS .67-64-1) present in any test concentration (75 µL/L).

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

The bluegill (SLS lot t88A9A) were obtained from a commercial supplier in Missouri. Prior to testing, these fish were held in a 500-L fiberglass tank under a photoperiod of 16 hours light and 8 hours darkness. The well water which flowed into this holding tank was characterized as having a total hardness and alkalinity range as calcium carbonate (CaC03 ) of 27 - 32 mg/L and 22 - 24 mg/L, respectively, and a specific conductance range of 100 - 130 micromhos per centimeter. Other parameters monitored in the holding tank inclUded pH with a range of 6.7 - 7.1, dissolved oxygen concentration ranging from of 75 - 80% saturation and a flow rate of 6.9 - 7.5 tank volume replacements/day (Weekly Record of Fish Holding Characteristics). Test fish were maintained under similar conditions for a minimum of 14 days prior to testing. The temperature range in the holding tank was 21 - 23°C during this 14day period. All fish were fed a commercial pelleted food, ad libitum, daily except during the 48 hours prior to testing. Fish were also fed on days 5, 9, 11 and 13 of the exposure. There was ≤ 0.1%' mortality in the test fish population during the two days prior to testing (Daily Record of Fi_h Holding Conditional. The bluegill used during this study were all of the same year class and a representative sample (n=30) of the fish from this population had a mean (range) wet weight of 1.7 (1.09 - 2.45) grams and total length of 50 (42 - 59) millimeters (Fish Weight and Lengths Log).

Test type:

flow-through

Water media type:

freshwater

Total exposure duration:

14 d

Test temperature:

21-23°C

pH:

6.9 - 7.6

Dissolved oxygen:

The protocol states that the total dissolved oxygen concentration is not allowed to drop below 6.6 mg/L (75% of saturation). During this study on two isolated occasions the percent saturation was 42% and 31% due to diluter malfunction On several other occasions during this study the dissolved oxygen fell below 75% of saturation but never dropped below 59% of saturation. Every aquaria was siphoned daily beginning on day five of the exposure period through test termination.

Nominal and measured concentrations:

Nominal test concentrations for the 14 days definitive study: 100, 65, 42, 27 and 18 µg A.I./L Anthraquinone
the mean measured test concentrations were 45, 34, 23, 16 and 12 µg A.I./L Anthraquinone.

Details on test conditions:

The test was conducted using an exposure system consisting of a constant flow serial diluter (Benoit et al, 1982), a t temperature controlled water bath, and a set of 14 exposure aquaria. The test system was designed to provide five concentrations of the test material, a dilution water control and a solvent control. The solvent control solutions contained the maximum amount of acetone (CAS 67-64-1) present in any test concentration (75 microL/L). All treatment levels and the controls were maintained in duplicate. Test aquaria were labelled to identify the test sample, nominal test concentration and designated replicate. Test solutions were not aerated. A photoperiod ot 16 hours light and 8 hours dark with a light intensity of 32 - 60 footcandles at the test solution surface was maintained throughout the test period. Lighting was provided by Durotest vita-LiteR fluorescent bulbs.
Each glass test aquarium measured 39 x 20 x 25 centimeters (cm) with a 19.5 cm high standpipe which maintained a constant test water volume of 15 L. The diluter was constructed to deliver 68 mL of solution per minute to each replicate test aquarium, providing approximately 6.5 volume replacements per aquarium every 24 hours. Test aquaria were impartially positioned in a water bath containing circulating water designed to maintain the test solution temperatures at 22 +/- 2°C.
The maximum organism loading concentration, (throughout the test), was 0.17 g of biomass per liter of flowing test solution per day.

Reference substance (positive control):

no

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 45 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

45 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

Following 14 days of exposure, there were no significant mortalities or adverse effects observed among bluegill exposed to any of the r.ean measured tested concentrations (45 - 12 µg A.I./L Anthraquinone). Based on this data it was established that under the test conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period), Anthraquinone was not acutely toxic to bluegill at and below the material's limit of water solubility. The 14-day LC50 for Anthraquinone and bluegill was empirically estimated as being greater than the highest mean measured test concentration (45 µg A.I·/L).

Conclusions:

Based on this data it was established that under the test conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period), Anthraquinone was not acutely toxic to bluegill at and below the material's limit of water solubility. The 14-day LC50 for Anthraquinone and bluegill was empirically estimated as being greater than the highest mean measured test concentration (45 µg A.I·/L).

Executive summary:

The purpose of this study was to estimate the acute toxicity of (LC50) of Anthraquinone to bluegill sunfish (Lepomis macrochirus) under flow-through conditions. In duplicate test aquaria, twenty organisms were exposed to five concentrations of Anthraquinone, a dilution water control and a solvent (acetone) control, via a flow-through system. Concentrations for this study were based on the limit of water solubility of Anthraquinone in freshwater, under the maintained test conditions. Prior to initiation of the definitive study, preliminary testing was conducted at Springborn Life Sciences, Inc., (SLS) to establish the water solubility and acute toxicity of Anthraquinone under test conditions. Test conditions for this study were generally consistent with the requirements published in the EPA´s Final Test Rule, Parts 704, 795 and 799 (Federal Register Volume 52, No. 107, 4 June 1987). Preliminary testing corroborated with information provided by the Mobay Corporation established that the limit of water solubility for Anthraquinone, under the maintained test conditions, was approximately 50µg A.I./L. During this same preliminary test period, it was also determined that concentrations of≤50µg/L Anthraquinone were not acutely toxic to bluegill sunfish. Based on these data, the following nominal concentrations were selected for the 14 day definitive study: 100, 65, 42, 27 and 18µg A.I./L Anthraquinone. Concentrations of the test material were maintained in the exposure vessels by introducing approximately 6.5 aquarium volumes per day of newly prepared teat solution via a constant flow serial diluter apparatus. Each replicate solution was sampled and analyzed for Anthraquinone concentration at least once prior to initiation, and on days 0 and 4 of the exposure period. Subsequently, on days 7 and 14, replicate solutions of the two highest test concentrations and the controls were sampled and analyzed. Based on the results of these analyses, the mean measured test concentrations were 45, 34, 23, 16 and 12µg A.I./L Anthraquinone. Throughout the exposure period a small amount of precipitate was observed in the diluter system's mixing chamber. However, no undissolved Anthraquinone (e.g., precipitate, film on solution's surface) was observed in any of the exposure vessels during the study. Biological observations were made and recorded at test initiation and every 24 hours thereafter until the test was terminated. Following 14-days of exposure there were no significant mortalities (i.e. > 10%) or adverse effects observed among bluegill at all treatment levels of Anthraquinone tested (45 - 12 µg A.I./L). Based on this data, it was established that under the conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period) Anthraquinone is not acutely toxic to bluegill sunfish at and below the material's limit of water solubility.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 - 29 July 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP, analytical monitoring

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

yes

Remarks:

The fish were fed during the final 48 hours before the test was initiated; the total alkalinity ranged from 19 - 22 mg/L CaCo3; the dissolved oxygen concentration ranged from 7.3 - 9.9 mg/L (71 - 96% of saturation); temperature ranged from 13 - I5°C.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

The high. middle and low test concentrations and the control solution were sampled and analyzed for Anthraquinone concentrations prior to the start of the definitive exposure. Results of these pretest analyses were used to judge whether sufficient quantities of test material were being delivered andmaintained in the exposure aquaria to initiate the definitive test. During the in-life phase of the definitive study, water samples were recoved from both replicate test solutions of each treatment level and the controls on test days 0 and 4 for analysis of Anthraquinone. Each exposure solution sample was collected from the approximate midpoint of the aquarium with a volumetric pipet. In addition, quality assurance (QA) blind samples were prepared at each sampling interval and remained with the set of exposure solution samples through the analytical process. These QA samples were prepared in dilution water at Anthraquinone concentrations unknown to the analyst. Results of the analyses of the QA sample were used to judge the precision and quality control maintained during the analysis ot exposure solution samples.

Vehicle:

yes

Details on test solutions:

A diluter stock solution of 1.34 .mg A.I./mL was prepared by diluting 0.3358 g of Anthraquinone (0.3352 grams of active ingredients) with acetone to a total volume of 250 mL. A Sage syringe pump in conjunction with a 50-mL gas-tiqht syringe was used to deliver the stock solution of Anthraquinone (1.34 mg A.I./mL) to the diluter's chemical mixing chamber containing approxiaately 1 liter of dilution water. The mixing chamber was positioned over a magnetic stirrer and inside of an ultrasonic water bath. The continuous stirring and sonication aided in the solubilization ot the test material. The solution contained in the mixing chamber constituted the highest nominal test concentration (100 µg A.l./L) and was subsequently diluted (65% dilution factor) to provide the desired exposure concentration range. A system similar to that used to deliver the test material was used to deliver acetone to the solvent control aquaria.
Tha solvant control solutions contained the maximum amount of acetone (CAS 67-64-1) present in any test concentration (75 µL/L).

Test organisms (species):

Oncorhynchus kisutch

Details on test organisms:

The coho salmon (SLS lot ,88A28) were obtained from a commerciai supplier in Washington. Prior to testing, these fish were held in a 500- L fiberglass tank under a photoperiod of 16 hours light and 8 hours darkness. The well water which flowed into this holding tank was characterized as having a total hardness and alkalinity range as calcium carbonate (CaC03) of 24 - 28 mg/L and 19 - 22 mq/L, respectively, and a specifc conductance range of 100 - 130 micromhos per centimeter. Other parameters monitored in the holdinq tank were a pH ranqe of 7.0 - 7.3. a dissolved oxygen concentration range of 68 - 83% of saturation and a flow rate of 9.6 - 13 tank volume replacements/day. Test fish were maintained under similar conditions for a minimum of 14 days prior to testing. The temperature range in the holding tank was 11 - 14°C during this 14day period. All fish were fed a dry commercial pelleted food, ad libitum, daily. There was 0.10% mortality in the test fish population durinq the two days prior to testing. The coho salmon used during this study were all of the same year class and a representative sampIer (n=30) of the fish from this population had a mean (range) wet weight of 1.75 (0.63-3.21) grams and total length of 59 (44 - 73) millimeters

Test type:

flow-through

Water media type:

freshwater

Total exposure duration:

96 h

Hardness:

19-22 mg/L CaCO3

Test temperature:

13 - 15°C

pH:

6.5 - 7.6

Dissolved oxygen:

7.3-9.9 mg/L (71 - 96 % of saturation)

Nominal and measured concentrations:

nominal test concentrations for the 96 hour definitive study: 100, 65, 42, 27 and 18 µg A.I./L Anthraquinone.
the mean measured test concentrations were 45, 30, 24, 18 and 12 µg A.l./L Anthraquinone.

Details on test conditions:

The test was conducted using an exposure system consisting of a constant flow serial diluter (Benoit et. ale 1982), a temperature controlled water bath, and set of 14 exposure aquaria. The test system was designed to provide five concentrations of the test matarial, one dilution water control and one solvent control.
All treataent levels and the controls were maintained in duplicate. Test aquaria were labelled to identify the test sample, nominal test concentration and designated replicate. Test solutions were not aerated. A photoperiod of 16 hours light and 8 hours darkness with a light intensity or 30 - 60
footcandles at the test solution surface was maintained throughout the test period. Lighting was provided by Vita-LiteR fluorescent bulbs.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 45 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

45 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

Following 96 hours of exposure, no mortalities or adverse effects were observed among coho salmon exposed to any of the tested concentrations (45 - 12 µg A.l./L Anthraquinone). Based on these data it was established that under the test conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period) Anthraquinone was not acutely toxic to coho Salmon at the material's limit of water solubility. The 96 hour LC50 for Anthraquinone and coho salmon was empirically estimated as being greater than the highest mean measured concentration of Anthraquinone tested (45 µg A.l./L).

Conclusions:

Based on these data it was established that under the test conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period) Anthraquinone was not acutely toxic to coho Salmon at the material's limit of water solubility.
The 96 hour LC50 for Anthraquinone and coho salmon was empirically estimated as being greater than the highest mean measured concentration of Anthraquinone tested (45 µg A.l./L).

Executive summary:

The purpose of this study was to estimate the acute toxicity (LC50 of Anthraquinone) to coho Salmon (Oncorhynchus kisutch) under flow-through conditions. Twenty organisms were exposed in duplicate test aquaria in a flow-through system to five concentrations of Anthraquinone, a dilution water control and a solvent (acetone) control. Selection of the nominal concentrations for this study was based on the limit of water solubility of Anthraquinone in freshwater under the maintained test conditions. Prior to initiation of the definitive study preliminary testing was conducted at Springborn Life Sciences, Inc., (SLS) to establish the water solubility and acute toxicity of Anthraquinone under test conditions. Test conditions for this study were generally consistent with the requirements published in the EPA's Final Test Rule, Parts 704, 795 and 799 (Federal Register Volume 52, No. 107, 4 June 1987). Preliminary testing corroborated information provided by the Mobay Corporation and established that the limit of water solubility for Anthraquinone, under the maintained test conditions, was approximately 50 µg/L. during this preliminary test period it was also determined that concentrations of ≤50 µg/L Anthraquinone were not acutely toxic to coho salmon. Based on these data the following nominal concentrations were selected for the 96 hour definitive study: 100, 65, 42, 27 and 18 µg A.I./L Anthraquinone. Concentrations of the test material were maintained in the exposure vessels by introducing approximately 6.5 aquarium volumes per day of newly prepared test solution via a constant flow serial diluter apparatus. Each replicate solution was sampled and analyzed for Anthraquinone concentration at test initiation and on day 4 of the exposure period. Baaed on the results of these analyses, the mean measured test concentrations were 45, 30, 24, 18 and 12 µg A.I./L Anthraquinone. Throughout the exposure period a small amount of precipitate was observed in the diluter system's mixing chamber; however no undissolved Anthraquinone (e.g., precipitate, film on solution's surface) was observed in any of the exposure vessels during the study. Biological observations were made and recorded at test initiation and every 24 hours thereafter until the test was terminated.

Following 96 hours of exposure, no mortalities or adverse effects were observed among coho salmon at all treatment levels of Anthraquinone tested (45 - 12 µg A.I./L). Based on these data, it was established that under the conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period) Anthraquinone is not acutely toxic to coho salmon at the material's limit of water solubility.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1-5 August 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP, analytical monitoring

Qualifier:

according to guideline

Guideline:

other: U.S. EPA 1985. Standard Evaluation Procedures for Acute Toxicity Test for Estuarine and Marine Organisms. Hazard Evaluation Division, Office of Pesticide Programms. Draft June 17, 1985.

Deviations:

yes

Remarks:

dissolved oxygen, temperature of test solutions, diluter

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

All test concentrations and the control solution were sampled and analyzed for Anthraquinone concentrations prior to the start of the definitive exposure. Results of these pretest analyses were used to jUdge whether sufficient quantities of test material were being delivered and maintained in the exposure aquaria to initiate the definitive test. During the in-life phase of the definitive study, water samples were removed from both replicate test solutions of each treatment level and the controls on test days 0 and 4 for analysis of Anthraquinone. Each exposure solution sample was collected from the approximate midpoint of the aquarium with a volumetric pipet. In addition, quality assurance (QA) blind samples were prepared at each sampling interval and remained with the set of exposure solution samples through the analytical process. These QA samples were prepared in dilution water at
Anthraquinone concentrations unknown to the analyst. Results of the analyses of the QA samples were used to judge the precision and quality control maintained dnring the analysis of exposure solution samples.

Vehicle:

yes

Details on test solutions:

A diluter stock solution of 1.34 mg A.I./mL was prepared by diluting 0.3358 g of Anthraquinone (0.3352 grams of active ingredient) with acetone to a total volume of 250 mL. A Sage syringe pump in conjunction with a SO-mL gas-tight syringe was used to deliver the stock solution of Anthraquinone (1.34 mg A.I./mL) to the diluter's chemical mixing chdmber containing approximately 1 liter of dilution water. The mixing chamber was positioned over a magnetic stirrer and inside of an ultrasonic water bath. The continuous stirring and sonication aided in the solubilization of the test material. The solution contained in the mixing chamber constituted the highest nominal test concentration (100 µg A.I./L) and was subsequently diluted (65% dilution factor) to provide the desired exposure concentration range. A syseem similar to that used to deliver the test material was used to deliver acetone to the solvent control aquaria.
The solvent control solutions contained the maxiama amount of acetone (CAS 67-64-1) present in any test concentration (75 µL/L).

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The rainbow trout were obtained from a commercial supplier in Montana. Prior to testing, these fish were held in a 500-L fiberglas aquaria, under a photoperiod of 16 hours light and 8 hours darkness. The well water which flowed into this holding tank was characterized as having a total hardness and alkalinity range as calcium carbonate (CaC03) of 24 - 28 mg/L and 19 - 22 mg/L, respectively, and a specific conductance range of 100 - 130 micrombos per centimeter (Weekly Gravity Feed Tank Water Quality Analysis Log Book). Other parameters monitored in the holding tank were a pH range of 6.9 - 7.1, a dissolved oxygen concentration range of 76 - 83% of saturation and a flow rate of 8.8 - 11.5 tank volume replaceaents/day (Weekly Record of Fish Holding Characteristics). Test fish were maintained under similar conditions for a minimum of 14 days prior to testing. The temperature range in the holding tank was 13 - 14°C during this 14day period. All fish were fed a dry ccmaercial palleted food, Ad libidum, daily. There was 0.4% mortality in the test fish population during the two days prior to testing. The rainbow trout used during this study were all of the same year class and a representative sample (n-30) of the fish from this populetion had a mean (range) wet wight of 1.49 (0.55-2.43) grams and total lenght of 55 (41 - 65) millimeters (Fish Weight and Lengths Log).

Test type:

flow-through

Water media type:

freshwater

Total exposure duration:

96 h

Details on test conditions:

The test was conducted using an exposure system containing of a constant flow serial diluter (Benoit et. al. 1982), a temperature-controlled water bath, and set of 14 exposure aquaria. The test system was designed to provide five concentrations of the test material, one dilution water control and
one solvent control. All treatment levels and the controls were maintained in duplicate. Test aquaria were labelled to identify the test sample, nominal test concentration and designated replicate. Test solutions were not aerated. A photoperiod of 16 hours light and 8 hours darkness with a light intensity of 30 - 60 footcandies at the test solution surface was maintained throughout the test period.
Each glass test aquarium measured 39 x 20 x 25 centimeters with a 19.5 cm high standpipe which maintained a constant test water volume of 15 L. The diluter was constructed to deliver 68 mL of solution per minute to each replicate test aquarium which provided approximately 6.5 volume replacements per aquarium every 24 hours. Test aquaria were impartially positioned in a water bath containing circulating water cooled by a Frigid Unit Chiller
designed to maintain the test solution temperatures at 12 ± 2°C.
At any given time during the test, the maximum organism loading concentration was 0.15 g of biomass per liter of flowing test solution per day.
The dilution water was from the same source as the water which flowed into the fish holding tank.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 55 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

Following 96 hours of exposure, no significant toxicant related mortalities (i.e, >10%) or adverse effects were observed among rainbow trout exposed to any of the tested concentrations (55 - 10 µg A.l./L Anthraquinone). Based on these data it was established that under the test conditions maintained during this study (e.g., under quality, solution temperature, solvent concentration, mixing period) Anthraquinone was not acutely toxic to rainbow trout at the material's limit of water solUbility. The 96 hours LC50 for Antllraquinone and rainbow trout was empirically estimated as being greater than the highest mean measured concentration of Anthraquinone tested (55 µg A.l./L).

Conclusions:

Based on these data it was established that under the test conditions maintained during this study (e.g., under quality, solution temperature, solvent concentration, mixing period) Anthraquinone was not acutely toxic to rainbow trout at the material's limit of water solubility.

Executive summary:

The purpose of this study was to estimate the acute toxicity (LC50) of Anthraquinone to rainbow trout (Salmo gairdneri) under flow-through conditions. Twenty organisms were exposed in duplicate test aquaria in a flow-through system to five concentrations of Anthraquinone, a dilution water control and a solvent (acetone) control. Selection of the nominal concentrations for this study was based on the limit of water solubility of Anthraquinone in freshwater under the maintained test conditions. Prior to initiation of the definitive study preliminary testing was conducted at Springborn Life Sciences, Inc., (SLS) to establish the water solubility and acute toxicity of Anthraquinone under test conditions. Test conditions for this study were generally consistent with the requirements published in the EPA'S Final Test Rule, Parts 704, 795 and 799 (Federal Register Volume 52, No. 107, 4 June 1987). Preliminary testing corroborated information provided by the Mobay Corporation and established that the limit of water solubility for Anthraquinone, under the maintained test conditions, was approximately 50µg/L. During this same preliminary test period it was also determined that concentrations of≤50µq/L Anthraquinone were not acutely toxic to coho salmon, a species similar to rainbow trout. Based on these data the following nominal concentrations were selected for the 96 hour definitive study: 100, 65, 42, 27 and 18µg A.I./L Anthraquinone. Concentrations of the test material were maintained in the exposure vessels by introducing approximately 6.5 aquarium volumes per day of newly prepared test solution via a constant flow serial diluter apparatus. Each replicate solution was sampled and analyzed for Anthraquinone concentration at test initiation and on day 4 of the exposure period. Based on the results of these analyses, the mean measured test concentrations were 55, 35, 23, 18 and 10µg A.I./L Anthraquinone. Throughout the exposure period a small amount of precipitate was observed in the diluter system's mixing chamber; however no undissolved Anthraquinone (e.g., precipitate, film on solution's surface) was observed in any of the exposure vessels during the study. Biological observations were made and recorded at test initiation and every 24 hours thereafter until the test was terminated.

Following 96 hours of exposure, no significant toxicant-related mortality or adverse effects were observed among rainbow trout at all treatment levels of Anthraquinone tested (55 - 10µg A.I./L).Based on these data, it was established that under the conditions maintained during this study (e.g., water quality, solution temperature, solvent concentration, mixing period) Anthraquinone is not acutely toxic to rainbow trout at the material's limit of water solubility.

Description of key information

Short-term toxicity to fish:
LC50 (96 h): > 45 µg/L for Oncorhynchus kisutch 
LC50 (14 d): > 45 µg/L for Lepomis macrochirus
LC50 (96 h): > 55 µg/L for Salmo gairdneri

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

45 µg/L

Additional information

Toxicity studies are waived because of high insolubility of anthraquinone in water.

There are three reliable acute toxicity studies for fish available, where selection of the test concentrations was based on the limit of water solubility of Anthraquinone in freshwater.

No significant mortalities or adverse effects were observed among fish at all treatment levels of Anthraquinone tested. Based on these data, it was established that under the conditions maintained during these studies (e.g., water quality, solution temperature, solvent concentration, mixing period) Anthraquinone is not acutely toxic to fish at the material's limit of water solubility.