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Benzenepropanoic acid,3,5-bis(1,1-dimethylethyl)-4-hydroxy-,C7-9 branched alkyl esters (CAS 125643-61-0) and Butyl 3,5 -bis(1,1-dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) are known as poorly water soluble substances, therefore, a modification of the standard method for the preparation of aqueous media was performed. An approach endorsed by several important regulatory authorities is to expose organisms to filtered Water Accommodated Fractions (filtered WAFs) of the test material in cases where the test material is a complex mixture and is poorly soluble in water and in the permitted auxiliary solvents and surfactants.

Short-term toxicity to fish

For the substance Benzenepropanoic acid, 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-C7 -9 branched alkyl esters (CAS 125643 -61 -0) no data is available.

Experimental results are available for read-across possibilities Benzenepropanoic acid 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-2 -ethyl-hexylester (CAS 144429 -84 -5) (key study) and Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) (key and supporting study).

Long-term toxicity to fish

Chronic toxicity to fish is not investigated so far for the substance Benzenepropanoic acid, 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0).

One experimental result is available for Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2). The study assess the effects of the substance on freshly hatched larvae of Pimpephales promelas (Goodband and Mullee, 2009). The mean hatching rate ranged from 93 % to 97% and the mean survival rate ranged from 80 % to 97 %, confirming that the substance is not of high concern for chronic toxicity to fish.

Short-term toxicity to aquatic invertebrates

The acute toxicity to invertebrates of Benzenepropanoic acid, 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0) as well as Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) and Benzenepropanoic acid 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-2 -ethyl-hexylester (CAS 144429 -84 -5) was investigated according to OECD guideline 202 and EU Method C.2 as static freshwater test with Daphnia magna used as test species.

For Benzenepropanoic acid,3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0) the following results are reported: EC50(48h) > 0.323 mg/L with a corresponding NOEC of 0.323 mg/L (based on peak area); Based on peak area attributed to test material: EC50(48h) > 0.224 mg/L, NOEC: 0.224 mg/L (Mead and Mullee, 2007). A certain potential for acute toxicity effetcts to daphnids cannot be excluded completely.

The study report for Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) gives only an Effective Loading rate concentration (EL50(48h): 110 mg/L loading rate WAF) as result with its corresponding No Observed Effect Loading rate (NOELR: 56 mg/L) (Wetton and Mullee, 2000).

No toxic effects at saturation were found in an experiment with Benzenepropanoic acid 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-2 -ethyl-hexylester (CAS 144429 -84 -5), which was tested as an aqueous preparation with a concentration of 100 % v/v saturated solution (Goodband and Mullee, 2009).

Long-term toxicity to aquatic invertebrates

Regarding chronic toxicity to aquatic invertebrates of Benzenepropanoic acid, 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0) neither experimental nor predicted results are available.

The substance Benzenepropanoic acid 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-2 -ethyl-hexylester (CAS 144429 -84 -5) was tested according to OECD guideline 211 (Goodband and Mullee, 2010). The following results were obtained: EL50(21d) > 100 mg/L (based on immobilisation, same for reproduction), LOELR: 10 mg/L, NOELR: 3.2 mg/L. The results were based on nominal loading rates only.

The same experiments were conducted with Butyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate (CAS 52449 -44 -2) (key studies, both reliability 1). Again, only Effective Loading rates are reported in the first study report (Wetton and Mullee, 2000). The second study was terminated on Day 8 due to 100 % mortality at all the test loading rate WAFs employed prior to the production of young daphnids (Goodband and Mullee, 2010).

Toxicity to aquatic algae and cyanobacteria

The growth inhibition effect of Benzenepropanoic acid, 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0) as well as Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) and Benzenepropanoic acid 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-2 -ethyl-hexylester (CAS 144429 -84 -5) on the green algae species Desmodesmus subspicatus was investigated according to OECD guideline 201 and EU Method C.3 as static freshwater tests.

The experiment with Benzenpropanoic acid, 3,5 -bis(1,1 -dimethylehtyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0) resulted in an Effective Loading rate (EL50) > 100 mg/L loading rate WAF with a corresponding No Observed Effect Loading rate of 100 mg/L (Mead and Hill, 2007).

In the study conducted with Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) the following results were reported: EL50 > 1000 mg/L and correspondingly the No Observed Effect Loading rate (NOELR) was 1000 mg/L.

The definitive test for Benzenepropanoic acid 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-2 -ethyl-hexylester (CAS 144429 -84 -5) showed that there should be a certain concern regarding toxicity to algae, when considering the following results: EC50 > 0.00018 mg/L, LOEC: 0.00018 mg/L (based on growth rate, yield and biomass integral), NOEC: 0.00014 mg/L (Vryenhoef and Mullee, 2009).

Toxicity to microorganisms

The respiration inhibition of Benzenepropanoic acid,3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-,C7 -9 branched alkyl esters (CAS 125643 -61 -0) on activated sewage sludge was investigated for a time period 3 hours and resulted in an EC50 > 1000 mg/L, with a corresponding NOEC of 1000 mg/L (Clarke, 2007). These results confirm that the substance shows no toxicity to aquatic microorganisms. The same can be concluded for Butyl 3,5 -bis(1,1 -dimethylethyl)-4 -hydroxybenzenepropanoate (CAS 52449 -44 -2) (Mead, 2000), Benzenepropanoic acid,3,5 -bis(1,1 -dimethylethyl)-4 -hydroxy-ethylhexylester (CAS 144429 -84 -5) (Clarke, 2007) and Vanlube BHC (Mead, 2007), due to the fact that all studies showed the same results as mentioned above. It was considered unnecessary and unrealistic to test at concentrations in excess of 1000 mg/L.