1-hydroxy-4-(p-toluidino)anthraquinone

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

see 'Principles of method if other than guideline'

Principles of method if other than guideline:

However, a preliminary range-finding test had identified the test substance to be of low toxicity (ie. the 96 hour LC50 was estimated as > 500 mg/L). The test substances which were of low toxicity were tested using a limit test at a concentration of 500 mg/L. A stock of 10 g/L was prepared and triplicate test concentrations prepared by adding 500 mL of stock to 9.5 litres of dilution water.
There were three other deviations from the general procedures. Firstly, the fish were transferred directly from their stock tank into the test solutions. Secondly, the temperature range during the holding/acclimation period was 12.9 to 16.3 °C and during the test ranged from 13.2 to 15.0°C. A third deviation was that the test concentrations were prepared and tested as "water-accommodated fractions" of Serip]as Blue 5RLN-TP. The procedures follow the OECD Guidelines for the Testing of Chemicals: Fish Acute toxicity Test Ref.203
The measured temperatures in the test tanks were outside the standard range of 14 to 16 °C during the test and ranged from 13.2 to 15.0 °C. However, this was not considered likely to have significantly affected the response of the fish.

GLP compliance:

yes

Specific details on test material used for the study:

- Purity not specified
- Chemical name: Seriplas Blue 5RLN-TP
- Binnie Environmental Ref.: 039418
- Supplied by: Yorkshire Chemicals plc
- Date of receipt: 10.03.1994
- Storage: Cool conditions
- Appearance: dull blue powder
- Odour: None
- Solubility in water: Insoluble

Analytical monitoring:

no

Details on sampling:

Effluents are stored at temperatures below 15 °C and are tested as soon after receipt as is practically possible.

Vehicle:

no

Details on test solutions:

A stock of 10 g/L was prepared and triplicate test concentrations prepared by adding 500 mL of stock to 9.5 litres of dilution water.

Preparation of Test Solutions
In cases where the test suhstance is soluble in water. each test solution is prepared by direct addition of the reyquired amount of test substance lo the dilution water. In cases where the substance has limited solubility then one of the following methods is used in order to maintain an adequate level of dispersion throughout the test period:
(i) acetone/methanol is used to disperse the test substance
(ii) a benign dispersing/emulsifying agent is used
(iii) the sample is dispersed by mechanical or other suitable apparatus, shielded from the test animals.
The method chosen depends upon the conccntrntion levels of the test substance required, and the reasons for a particular choice are discussed in the main body of the report.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

- Supplier: Upwey Trout Farm, Weymouth, Dorset
- Date of arrival at laboratory: 1994-03-09
- Holding/ Acclimation period: 1994-03-09/1994-03-28
- Acclimation conditions: 12.9-16.3 °C; Dissolved oxygen: > 80% air saturation value
- Food: diet of Trout Fry feed (BP Nutrition (U.K.) Limited)
- Keeping water: dechlorinated mains water
- Mean length of fish: 51.6 mm
- Mean weight of fish: 1.18 g
- The sizes of the animals were found to be in approximately normal (Gaussian) distribution

The rainhow trout are examined prior to selection for the test to ensure that only heatlthy animals are exposed. Following the completion of the test a randum sample of 10 animals is measured and weighed, and the results plotted on a linear/prohit scale in order to check that they are in an approximately normal distribution.
The animals are held in the lahoratory for at least twelve days befrore the start of the test and they are fed on a diet of Trout Fry feed manufactured hy BP Nutrition (U.K.) Limited. Their environment is dechlorinated main water and the concentration of dissolved oxygen is maintained close to saturation by continuously diffusing air through the water.

Test type:

semi-static

Water media type:

freshwater

Remarks:

dechlorinated mains water of typical hardness 240 mg calcium carbonate per liter

Limit test:

yes

Total exposure duration:

96 h

Hardness:

13.4 °dH = 240 mg/L CaCO3 (confirmed analytically by titration with EDTA solution.

Test temperature:

13.2 - 15.0 °C

pH:

7.6 - 8.5

Dissolved oxygen:

91-100 % ASV (air saturation value)

Nominal and measured concentrations:

Water Accommodated Fraction (WAF) was used to test effects at effective loading of 500 mg/L because the substance is insoluble in water.

Details on test conditions:

- Test vessels: plastic aquaria with a working volume of 10 litre
- Experimental design: 1 test concentration plus 1 control
- Number of fish: 10 per test concentration
- Feeding during exposure: None
- Method of initiation: fish were placed in prepared media
- Loading: 1.18 g (+/-0.21) body weight
- Photoperiod: 16 h light : 8 h dark
- Temperature: 14 - 16 °C
- Aeration: gentle aeration
- Test concentration/s: 500 mg/L
- Replicates: 3 (test concentration), 1 (control)
- Method of administration: direct weighing
- Medium renewal: daily
- Monitoring data: pH, dissolved oxygen and temperature were measured and recorded daily
- Criteria of effects: absence of response to physical stimulation; in addition to observations on mortality the first 6 hours and thereafter at 24, 48, 72 and 96 hours, type and incidence of sub-lethal effects compared with control fish were observed

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The measured temperatures in the test tanks were outside the standard range of 14 to 16 °C during the test and ranged from 13.2 to 15.0 °C. However, this was not considered likely to have significantly affected the response of the fish.

Sublethal observations / clinical signs:

Test substances identified by preliminary studies as being of low toxicity (ie. where the 96 hour LC50 > 500 mg/L) are typically assessed further by using a limit test at a concentration of 500 mg/L. To determine effects accurately, limit tests use three test vessels each containing ten fish at the single test substance concentration. The study reported here resulted in no mortalities for thirty fish exposed to a 500 mg/L concentration of Solvent Violet 13. The limit test therefore confirmed the preliminary test results that the 96 hour LC,0 was greater than 500 mg/L of Solvent Violet 13.

Solvent Violet 13 is described by Yorkshire Chemicals' Product Safety Data Sheet as "insoluble" in water. The test concentrations reported in this study can therefore only be described as "water-accommodated fractions". However, the data reported would give a true reflection of the aquatic tnxicity of Solvent Violet 13 and can be considered an

environmentally realistic testing approach since it reflects what would happen to a sensitive fish species if the test substance were to enter or contaminate a body of freshwater.

The measured temperatures in the test tanks were outside the standard range of 14 to 16°C during the test and ranged from 13.2 to 15 .0°C. However, this was not considered likely to have significantly affected the response of the fish.

Validity criteria fulfilled:

yes

Remarks:

mortality in the controls < 10% by the end of the test; dissolved oxygen concentration above 60% of the air-saturation value throughout the exposure period; pH did not vary by more than 1 unit

Conclusions:

The acute toxicity of Solvent Violet 13 to fish was conducted in a static test according to the OECD Guideline 203. After 96 hours of exposure a LC50 value of higher than 500 mg/L was obtained.

Executive summary:

The objective of the test is lo determine the 96 hour median lethal concentration (96 huur LCw) of Solvent Violet 13 to rainbow trout (Oncorhynchus mykiss).

The test was carried out in accordance with OECD Guideline 203, 'Fish Acute Toxicity Test'. A preliminary range-finding test had identified the Solvent Violet 13 to be of low toxicity (ie. the 96 hour LC50 was estimated as > 500 mg/L). Therefore the main test was carried out under semi static conditions at a limit test concentration of 500 mg/L. A stock of 10 g/L was prepared and triplicate test concentrations prepared by adding 500 mL of stock to 9.5 litres of dilution water. The fish were transferred directly from their stock tank into the test solutions and the temperature range during the holding/acclimation period was 12.9 to 16.3°C and during the test ranged from 13.2 to 15.0 °C. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 6, 24, and 96 hours of exposure. After 96 hours no toxic effects of Solvent Violet 13 against fish were observed at 500 mg/L. This toxicity study is classified as acceptable and satisfies the guideline requirements for the acute fish study.

Description of key information

The test was carried out in accordance with OECD Guideline 203, 'Fish Acute Toxicity Test'. The main test was carried out under semi static conditions at a limit test concentration of 500 mg/L. A stock of 10 g/L was prepared and triplicate test concentrations prepared by adding 500 mL of stock to 9.5 litres of dilution water. The fish were transferred directly from their stock tank into the test solutions and the temperature range during the holding/acclimation period was 12.9 to 16.3°C and during the test ranged from 13.2 to 15.0 °C. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 6, 24, and 96 hours of exposure. After 96 hours no toxic effects of Solvent Violet 13 against fish were observed at 500 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

500 mg/L

Additional information

"Should read: > 500 mg/L"