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Description of key information

Biodegradation in water

28-days Closed Bottle test following the OECD guideline 301 D was performed to determine the ready biodegradability of the test item. The study was performed at a temperature of 20°C. The test system included control, test item and reference item. Polyseed were used for this study. 1 polyseed capsule were added in 500 ml D.I water and then stirred for 1 hour for proper mixing and functioning of inoculum. This gave the bacterial count as 10E7 to 10E8 CFU/ml. At the regular interval microbial plating was also performed on agar to confirm the vitality and CFU count of microorganism. The concentration of test and reference item (Sodium Benzoate) chosen for both the study was 4 mg/L, while that of inoculum was 32 ml/L . OECD mineral medium was used for the study. ThOD (Theoretical oxygen demand) of test and reference item was determined by calculation. % degradation was calculated using the values of BOD and ThOD for test item and reference item. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 75.3 %. Degradation of Sodium Benzoate exceeds 49.39 % on 7 days & 75.3 on 14th day. The activity of the inoculum is thus verified and the test can be considered as valid. The BOD28 value of test chemical was observed to be 0.77 mgO2/mg. ThOD was calculated as 0.78 mgO2/mg. Accordingly, the % degradation of the test item after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 98.71 %. Based on the results, the test item, under the test conditions, was considered to be readily in nature.

Biodegradation in water and sediment

Estimation Programs Interface (EPI Suite, 2017) prediction model was run to predict the half-life in water and sediment for the test chemical. If released in to the environment, 31.3% of the chemical will partition into water according to the Mackay fugacity model level III and the half-life period of test chemical in water is estimated to be 37.5 days (900 hrs). The half-life (37.5 days estimated by EPI suite) indicates that the chemical is not persistent in water and the exposure risk to aquatic animals is moderate to low whereas the half-life period of test chemical in sediment is estimated to be 337.5 days (8100 hrs). However, as the percentage release of test chemical into the sediment is less than 1% (i.e, reported as 0.083%), indicates that test chemical is not persistent in sediment.

Biodegradation in soil

The half-life period of test chemical in soil was estimated using Level III Fugacity Model by EPI Suite version 4.1 estimation database (EPI suite, 2017). If released into the environment, 68.6% of the chemical will partition into soil according to the Mackay fugacity model level III. The half-life period of test chemical in soil is estimated to be 75 days (1800 hrs). Based on this half-life value of test chemical, it is concluded that the chemical is not persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to low

Additional information

Biodegradation in water:

Various experimental key and supporting studies for the target compound were reviewed for the biodegradation end point and their results are summarized as below:

The first study was experimental study in this study the 28-days Closed Bottle test following the OECD guideline 301 D was performed to determine the ready biodegradability of the test item. The study was performed at a temperature of 20°C. The test system included control, test item and reference item. Polyseed were used for this study. 1 polyseed capsule were added in 500 ml D.I water and then stirred for 1 hour for proper mixing and functioning of inoculum. This gave the bacterial count as 10E7 to 10E8 CFU/ml. At the regular interval microbial plating was also performed on agar to confirm the vitality and CFU count of microorganism. The concentration of test and reference item (Sodium Benzoate) chosen for both the study was 4 mg/L, while that of inoculum was 32 ml/L. OECD mineral medium was used for the study. ThOD (Theoretical oxygen demand) of test and reference item was determined by calculation. % degradation was calculated using the values of BOD and ThOD for test item and reference item. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 75.3 %. Degradation of Sodium Benzoate exceeds 49.39 % on 7 days & 75.3 on 14th day. The activity of the inoculum is thus verified and the test can be considered as valid. The BOD28 value of test chemical was observed to be 0.77 mgO2/mg. ThOD was calculated as 0.78 mgO2/mg. Accordingly, the % degradation of the test item after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 98.71 %. Based on the results, the test item, under the test conditions, was considered to be readily in nature.

In next experimental key study from peer reviewed journal (GREIM H, et. al; 1994), biodegradation experiment was conducted for 28 days for evaluating the percentage biodegradability of test chemical. The study was performed according to OECD Guideline 302 B (Inherent biodegradability: Zahn-Wellens/EMPA Test). The percentage degradation of test chemical was determined to be >70% by DOC removal or COD parameter in 28 days. Thus, based on percentage degradation, test chemical is considered to be readily biodegradable in water.

Another biodegradation study was conducted for 28 days for evaluating the percentage biodegradability of test chemical (IUCLID dataset, 2000). The study was performed according to OECD Guideline 302 B "Inherent biodegradability: Modified Zahn–Wellens Test" under aerobic conditions. The percentage degradation of test chemical was determined to be >90% by DOC removal or COD parameter in 28 days. Thus, based on percentage degradation, test chemical is considered to be readily biodegradable in nature.

In next supporting study from secondary source (IUCLID dataset, 2000), biodegradation experiment was conducted for 28 days for evaluating the percentage biodegradability of test chemical. The study was performed according to "Modified Zahn–Wellens Test" under aerobic conditions. Activated sludge was used as a test inoculum for the study. Initial test substance conc. used in the study was 200 mg/l based on Dissolved Organic Carbon (DOC), respectively. Lag phase and beginning of the plateau phase was determined to be 1 and 3 days, respectively. The percentage degradation of test chemical was determined to be 100% by DOC removal or COD parameter after 5 days. Thus, based on percentage degradation, of test chemical is considered to be readily biodegradable in nature.

Additional biodegradation study from authoritative database (J-CHECK and Envichem, 2017) and secondary source (IUCLID dataset, 2000) was conducted for 14 days for evaluating the percentage biodegradability of test chemical. The study was performed according to OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I) under aerobic conditions. Activated sludge (non-adapted) was used as a test inoculums for the study. Concentration of inoculum i.e, sludge used was 30 mg/l and initial test substance conc. used in the study was 100 mg/l, respectively. The percentage degradation of test chemical was determined to be 0, 4.6 and 1% by BOD, TOC removal and UV-Vis parameter in 14 days. Thus, based on percentage degradation, test chemical is considered to be not readily biodegradable in nature.

In an another supporting study, biodegradation experiment was conducted for 6 hrs for evaluating the percentage biodegradability of test chemical by using acclimated activated sludge as a test inoculum (Sarvottam D. Deshpande, et. al; 1987). Activated sludge obtained from an oxidation ditch treating domestic waste water was used as a test inoculum for the study. For acclimation of activated sludge, activated sludge was exposed to test chemical. The organic loading in each case was maintained at 0.11 g of total organic carbon (TOC)/g of mixed liquor volatile suspended solids (MLVSS). Diammonium orthophosphate (200 mg/L) provided the required nitrogen and phosphorus. The fill and draw technique was employed, and acclimation was determined from the rate of TOC utilization by the respective sludges. Biodegradation experiment was carried out by exposing the activated sludge to the test chemical.The organic loading in each case was maintained at 0.11 g of total organic carbon (TOC)/g of MLVSS in batch reactors of 1 l capacity and equipped with sintered glass diffusers. The experiments were conducted for a period of 6 h, and the TOC of 0- and 6-h samples in each case was monitored with a Beckman TOC analyzer (Model 915 A). The organic components present in samples were quantified with the help of high-performance liquid chromatography (HPLC) (Model 204, Water Associates Inc., Milford, MA) in a system equipped with M-6000 and M-45 pumps, a u6K septumless injector, a Model 440 UV absorption detector with a 254-nm primary filter, and an HPLC recorder (Houston Instrument Series B-5000 dual pen, Austin, TX). Solvents used were double-distilled spectroscopic-grade dioxane and water in the ratio of 1:24 in the case of catechol-resorcinol system and only water in the case of all other systems investigated. The column used for HPLC analyses was µBondapac C-18. Samples filtered through Millipore membrane filter were analysed by reverse-phase chromatography. Operating conditions were (i) chart speed, 25cm/h; (ii) flow rate, 2mL/min; and (iii) column temperature, ambient. Analytical-grade standards were run with each experiment. The TOC removal of the test chemical by using acclimated activated was determined to be 16.9 mg after a period of 6 hrs. Thus, based on this, test chemical is considered to be biodegradable in nature.

 On the basis of above results for target (from study report and peer reviewed journals and secondary source), most of the studies show test chemical is readily biodegradable therefore it can be concluded that the test chemical is readily biodegradable in nature.

Biodegradation in water and sediment

Estimation Programs Interface (EPI Suite, 2017) prediction model was run to predict the half-life in water and sediment for the test chemical. If released in to the environment, 31.3% of the chemical will partition into water according to the Mackay fugacity model level III and the half-life period of test chemical in water is estimated to be 37.5 days (900 hrs). The half-life (37.5 days estimated by EPI suite) indicates that the chemical is not persistent in water and the exposure risk to aquatic animals is moderate to low whereas the half-life period of test chemical in sediment is estimated to be 337.5 days (8100 hrs). However, as the percentage release of test chemical into the sediment is less than 1% (i.e, reported as 0.083%), indicates that test chemical is not persistent in sediment.

Biodegradation in soil

The half-life period of test chemical in soil was estimated using Level III Fugacity Model by EPI Suite version 4.1 estimation database (EPI suite, 2017). If released into the environment, 68.6% of the chemical will partition into soil according to the Mackay fugacity model level III. The half-life period of test chemical in soil is estimated to be 75 days (1800 hrs). Based on this half-life value of test chemical, it is concluded that the chemical is not persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to low.

On the basis of available information, the test chemical can be considered to be readily biodegradable in nature.