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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Description of key information

The LC50 for Fluorescein sodium was found to be 337 (278±403) mg/liter for Daphnia pulex neonates on the basis of the results obtained from mortality and the response predicted by regression analysis. 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
337 mg/L

Additional information

Various studies for the target chemical fluorescein sodium (518-47-8)along with its read across substance were reviewed to summarize the following information: 

The toxicity of dye, fluorescein sodium salt, to Daphnia pulex was assessed using acute level toxicological test (W.K. Walthall, J.D. Stark). The test organisms were obtained from cultures at or beyond the third filial (F3) generation. The final concentrations used in the generation of the acute mortality regression are 0, 150, 250, 400, 600 and 800 mg/liter. No control mortality occurred in the acute toxicity experiments. For each concentration tested, 25 ml of test solution was transferred into a 30-ml plastic cup and five neonates were transferred into the test container using a disposable glass pipette. Each concentration was replicated a minimum of four times. The testing regime was 48 h static, nonrenewal. The LC50for Fluorescein sodium was found to be 337 (278±403) mg/liter for Daphnia pulex neonates on the basis of the results obtained from mortality and the response predicted by regression analysis.


Short term toxicity study was conducted (Kelly A. Stockton, Christine M. Moffitt) to determine the aquatic toxicity of sodium fluorescein.The dose concentration used are0.2, 20.0, 50.0 71.0 100.0 141.0 and 200.0mg/L. The snail behavior was observed every 15 minutes during the first hour of exposure, and then every hour for the next 4 h. At the end of a 24h exposure, we poured the test system into small sieve to recover all snails, and then rinsed the snail three times with clean, aged laboratory water. The snails are then placed s into small cups with aged water, assessed mortality immediately, then again at 24 and 48 h. Each beaker was inspected with the aid of a dissecting microscope and watched for snail movement, and touched individual snails that were not active with a probe to elicit movement or tactile response. Each trial was replicated three times over two weeks for a total of 9 beakers tested at each concentration. Probit model and normal distribution were used for estimating the probability of mortality. Based on the experiment the median lethal concentration was determined to be 377 mg/L for snails.

Ecological assessments (Malcolm S. Field, Ronald G. Wilhelm, James F. Quinlan, and Thomas J. Aley) included both acute and chronic effects for the 3 life forms: fish, Cladocera daphnia, and algae. The concern levels identified by use of the SAR technique are expressed as high, moderate and low. These values are toxicity values, not exposure values. SAR results are generally regarded by EPA as being reliable.

Dyes with aquatic toxicity values below 1mg/l received a high concern rating, those with toxicity values ranging from 1-100mg/l received a moderate concern rating and those with toxicity values greater than 100mg/l received a low concern rating.

The Median Lethal Concentration which is estimated to be lethal to 50% of Cladocera (water flea) in 48 hours exposure to the test chemical (LC50) was found to be 165mg/l.


Toxicity tests (A marine toxicity assessment of selected tracer dyes using standard bioassays )on the tracer dyes (Fluorescein)

were carried out using standard bioassay using the Copepod acute toxicity test .The test were conducted according to ISO (1999) Guidelines - ISO 14669 Water Quality - Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea). These bioassays were carried out in 12 well polystyrene plates, each well contained 5mls of test solution and 5 T.battagliaicopepodites (approximately 4-6 days old). There were 4 replicates per concentration and the test was carried out over a period of 48 hours. The Tisbe were observed using a binocular microscope and mortality recorded. All parameters were within acceptable limits. A 48 h Zinc reference study was carried for quality assurance purposes alongside the tracer dyes to test the sensitivity of the species population for validity of the study. The results showed that EC50 for Fluorescein was determined to be 160.7 mg/l and the chemical was considered as not classified according to the CLP regulations.

Based on the studies on fluorescein sodium and read across, the test chemical was considered as a not classified to short term toxicity to aquatic invertebrates and also according to the CLP regulations.