Tetrabutylammonium bromide

Endpoint summary

Description of key information

Bioaccumulation:

Bioaccumulation study in fish was conducted for determining the bioconcentration factor of test chemical. The study was performed in accordance with the OECD Guideline 305 (Bioaccumulation in Fish: Aqueous and Dietary Exposure) -I: Aqueous Exposure Bioconcentration Fish Test and EPA OPPTS 850.1730 (Fish Bioconcentration Test), respectively. Danio rerio (Zebra fish) (male) of total length 2.5 to 3.2 cm obtained from commercial fish farm, Manimangalam was used as a test fishes. Fish used in the tests were free from observable diseases and abnormalities. Test fishes were fed daily throughout the study period. Fishes were fed with an appropriate diet of known lipid and total protein content in an amount sufficient to keep them in a healthy condition and to maintain body weight. Fish species identification was done externally at Fisheries College and Research Institute (FCRI), Ponneri, Tiruvallur District, Tamil Nadu. Acclimatization of the stock population of test fishes were done for 2 weeks. Test conditions were same as the test. Feeding to test fishes were done in the same manner as that done during the study. Preliminary study was carried out. A limit test was conducted at 100 mg/l alongwith control for 96 hrs. 7 fishes were exposed to 100 mg/l for a period of 96 hrs. An equal control was also maintained. No mortality or toxicity signs were observed during the study period upto 96 hrs. The highest conc. (1 mg/l) of the test chemical was selected base on 1% of its acute LC50 (100 mg/l). The second conc. (0.1 mg/l) was selected from the one above by factor of 10. Thus, exposure conc. were 0.1 and 1.0 mg/l, respectibely. The blended water was used in the study. It was prepared by mixing the reverse osmosis water and well water in the ratio of 3:1. This water was used during the acclomatization, preliminary experiment, uptake and depuration phase. The pH, hardness, total solids, total organic carbon (TOC), anions, cations, heavy metals and the presence of pesticides were analyzed.Glass aquaria (glass tank) of 30 lit capacity was used as a test vessel. Test chemical conc. used for the study were 0.1 and 1.0 mg/l. Total 72 fishes/vessel for each test group were exposed with the test chemical. Study was performed under semi-static conditions and test medium was renewed thrice in a week and durng the renewal. fish from each group was transferred to a new medium in which test conc. are exposed in a test chamber. Biomass loading rate during the study was 0.1 to 1.0 g/l, respectively. One control was run in addition to the test series. A control group of fish was held under experimental conditions except for the absence of the test chemical. The leftout food and faeces were siphoned daily from the test chambers shortly after feeding (30 mins to 1 hr). Test chambers were kept clean as possible throughout the test to keep the conc. of organic matter as low as possible to avoid the presence of organic carbon that may limit the bioavailability of the test chemical. The blended water was used in the study. It was prepared by mixing the reverse osmosis water and well water in the ratio of 3:1. This water was used during the acclomatization, preliminary experiment, uptake and depuration phase. The pH, hardness, total solids, total organic carbon (TOC), anions, cations, heavy metals and the presence of pesticides were analyzed. Test conditions involve a pH of 7.39 to 7.97, hardness of 139 to 140 mg/l as CaCO3, temperature of 23.4 to 25°C, dissolved oxygen of 84.3 to 102.7%, TOC of < 5 mg/l, respectively. Uptake phase was carried out for 28 d and depuration phase for 14 d, respectively. Photoperiod during the uptake phase was 12: 12 light:dark conditions controlled by an automatic timer and depuration phase was 6:6 light:dark conditions, respectively. All experiments were performed in triplicates.The leftout food and faeces were siphoned daily from the test chambers shortly after feeding (30 mins to 1 hr). Test chambers were kept clean as possible throughout the test to keep the conc. of organic matter as low as possible to avoid the presence of organic carbon that may limit the bioavailability of the test chemical. After removal of lipid content from the sample, the aqueous layer was transferred into 50 mL centrifuge tube and the volume was adjusted to 10 mL using acetonitrile. The sample was homogenized for 2 minutes using tissumizer. The sample solution was centrifuged at 3000 rpm and decanted, A 2 mL of sample solution was filtered through 0.2 um PTFE and analysed for active content. The final solution was filtered and analyzed by LC-MS/MS conditions. HPLC system consists of Agilent 1290 Infinity UHPLC, Agilent Zorbax Eclipse XDB (2.1 mm X 75 mm, 3.5 μm), column temperature of 40°C, injection volume of 10 μl, flow rate of 1.0 ml/min, auto dampler temp. of 10°C. Gradient conditions involve the use of 0.1% acetic acid in 90/10 (MilliQ water/ Methanol) and 0.1%  acetic acid in acetonitrile, respectively. Total run time was 9.0 mins with a retention time of 1.5 mins. MS conditions involves the detector of Agilent 6490 LC-MS/MS System, coupled with Mass Hunter instrument control and data processing software, AJS-ESI interface, Positive polarity, Gas temperature of 260°C, Gas flow of 16 L/min, Nebulizer gas flow of 50 Psi, Sheath gas temperature of 400°C, Sheath gas flow of 11 L/min, Capillary voltage of 3500V, CAV of 4 V and Dwell time of 200 sec., respectively. During the uptake and depuration phase, the fish samples were extracted immediately and analyzed for active content. The uptake phase rate constants (k1), depuration phase rate constant (k2) was calculated by plotting the Ln (comc. of fish, mg/kg) Vs occasions (days). During the acclimatization period, there was no mortaliy observed and fish was found to be healthy. In preliminary test: there was no mortalirty or toxicity signs in test fishes. During the study, the water temperature variation was less than + 2°C which ranged from 23.4 to 25.0°C, the concentration of dissolved oxygen had not been fallen below 60% saturation and the range was 84.3 to 102.7%, the concentration of the test item (0.1 & 1.0 mg/L) in the chambers was maintained within + 20% of the mean of the measured values during the uptake phase, the concentration of the test item (0.1 & 1.0 mg/L) was below its limit of solubility in water, taking into account the effect that the test water may have on effective solubility, no mortality and other adverse effect was found in both control and treated fish during the uptake and depuration phases. No significant differences in average growth between the test and the control groups of sampled fish (Hence there was no indication of toxic effect of the test chemical, thereby fulfilling the validity criteria and hence, study was considered to be valid.The Bio-concentration factor (BCF) was determined as 0,8394 and 0.4624 for 0.1 mg/L and 1.0 mg/L concentrations respectively in the uptake phase. Further, based on the overall rate constants of the uptake and depuration phases, the kinetic Bio-concentration factor (BCFk) was determined to be -0.1261 and -0.3114 for the concentrations 0.1 mg/L and 1.0 mg/L respectively. From the growth rate analysis, the growth corrected kinetic bio-concentration factor (BCFkg) was determined to be -0.1240 and -0.3173 for the concentrations 0.1 mg/L and 1.0 mg/L respectively. Based on the lipid content analysis, the lipid normalized kinetic bio-concentration factor (BCFkl) was determined to be -0.1852 and -0.5097 for the concentrations 0.1 mg/L and 1.0 mg/L respectively in the uptake phase. Since, the BCF value did not exceeded the threshold limit of 2000, thus, test chemical was considered to be non-bioaccumulative,

Additional information

Bioaccumulation:

Bioaccumulation study in fish was conducted for determining the bioconcentration factor of test chemical. The study was performed in accordance with the OECD Guideline 305 (Bioaccumulation in Fish: Aqueous and Dietary Exposure) -I: Aqueous Exposure Bioconcentration Fish Test and EPA OPPTS 850.1730 (Fish Bioconcentration Test), respectively. Danio rerio (Zebra fish) (male) of total length 2.5 to 3.2 cm obtained from commercial fish farm, Manimangalam was used as a test fishes. Fish used in the tests were free from observable diseases and abnormalities. Test fishes were fed daily throughout the study period. Fishes were fed with an appropriate diet of known lipid and total protein content in an amount sufficient to keep them in a healthy condition and to maintain body weight. Fish species identification was done externally at Fisheries College and Research Institute (FCRI), Ponneri, Tiruvallur District, Tamil Nadu. Acclimatization of the stock population of test fishes were done for 2 weeks. Test conditions were same as the test. Feeding to test fishes were done in the same manner as that done during the study. Preliminary study was carried out. A limit test was conducted at 100 mg/l alongwith control for 96 hrs. 7 fishes were exposed to 100 mg/l for a period of 96 hrs. An equal control was also maintained. No mortality or toxicity signs were observed during the study period upto 96 hrs. The highest conc. (1 mg/l) of the test chemical was selected base on 1% of its acute LC50 (100 mg/l). The second conc. (0.1 mg/l) was selected from the one above by factor of 10. Thus, exposure conc. were 0.1 and 1.0 mg/l, respectibely. The blended water was used in the study. It was prepared by mixing the reverse osmosis water and well water in the ratio of 3:1. This water was used during the acclomatization, preliminary experiment, uptake and depuration phase. The pH, hardness, total solids, total organic carbon (TOC), anions, cations, heavy metals and the presence of pesticides were analyzed.Glass aquaria (glass tank) of 30 lit capacity was used as a test vessel. Test chemical conc. used for the study were 0.1 and 1.0 mg/l. Total 72 fishes/vessel for each test group were exposed with the test chemical. Study was performed under semi-static conditions and test medium was renewed thrice in a week and durng the renewal. fish from each group was transferred to a new medium in which test conc. are exposed in a test chamber. Biomass loading rate during the study was 0.1 to 1.0 g/l, respectively. One control was run in addition to the test series. A control group of fish was held under experimental conditions except for the absence of the test chemical. The leftout food and faeces were siphoned daily from the test chambers shortly after feeding (30 mins to 1 hr). Test chambers were kept clean as possible throughout the test to keep the conc. of organic matter as low as possible to avoid the presence of organic carbon that may limit the bioavailability of the test chemical. The blended water was used in the study. It was prepared by mixing the reverse osmosis water and well water in the ratio of 3:1. This water was used during the acclomatization, preliminary experiment, uptake and depuration phase. The pH, hardness, total solids, total organic carbon (TOC), anions, cations, heavy metals and the presence of pesticides were analyzed. Test conditions involve a pH of 7.39 to 7.97, hardness of 139 to 140 mg/l as CaCO3, temperature of 23.4 to 25°C, dissolved oxygen of 84.3 to 102.7%, TOC of < 5 mg/l, respectively. Uptake phase was carried out for 28 d and depuration phase for 14 d, respectively. Photoperiod during the uptake phase was 12: 12 light:dark conditions controlled by an automatic timer and depuration phase was 6:6 light:dark conditions, respectively. All experiments were performed in triplicates.The leftout food and faeces were siphoned daily from the test chambers shortly after feeding (30 mins to 1 hr). Test chambers were kept clean as possible throughout the test to keep the conc. of organic matter as low as possible to avoid the presence of organic carbon that may limit the bioavailability of the test chemical. After removal of lipid content from the sample, the aqueous layer was transferred into 50 mL centrifuge tube and the volume was adjusted to 10 mL using acetonitrile. The sample was homogenized for 2 minutes using tissumizer. The sample solution was centrifuged at 3000 rpm and decanted, A 2 mL of sample solution was filtered through 0.2 um PTFE and analysed for active content. The final solution was filtered and analyzed by LC-MS/MS conditions. HPLC system consists of Agilent 1290 Infinity UHPLC, Agilent Zorbax Eclipse XDB (2.1 mm X 75 mm, 3.5 μm), column temperature of 40°C, injection volume of 10 μl, flow rate of 1.0 ml/min, auto dampler temp. of 10°C. Gradient conditions involve the use of 0.1% acetic acid in 90/10 (MilliQ water/ Methanol) and 0.1%  acetic acid in acetonitrile, respectively. Total run time was 9.0 mins with a retention time of 1.5 mins. MS conditions involves the detector of Agilent 6490 LC-MS/MS System, coupled with Mass Hunter instrument control and data processing software, AJS-ESI interface, Positive polarity, Gas temperature of 260°C, Gas flow of 16 L/min, Nebulizer gas flow of 50 Psi, Sheath gas temperature of 400°C, Sheath gas flow of 11 L/min, Capillary voltage of 3500V, CAV of 4 V and Dwell time of 200 sec., respectively. During the uptake and depuration phase, the fish samples were extracted immediately and analyzed for active content. The uptake phase rate constants (k1), depuration phase rate constant (k2) was calculated by plotting the Ln (comc. of fish, mg/kg) Vs occasions (days). During the acclimatization period, there was no mortaliy observed and fish was found to be healthy. In preliminary test: there was no mortalirty or toxicity signs in test fishes. During the study, the water temperature variation was less than + 2°C which ranged from 23.4 to 25.0°C, the concentration of dissolved oxygen had not been fallen below 60% saturation and the range was 84.3 to 102.7%, the concentration of the test item (0.1 & 1.0 mg/L) in the chambers was maintained within + 20% of the mean of the measured values during the uptake phase, the concentration of the test item (0.1 & 1.0 mg/L) was below its limit of solubility in water, taking into account the effect that the test water may have on effective solubility, no mortality and other adverse effect was found in both control and treated fish during the uptake and depuration phases. No significant differences in average growth between the test and the control groups of sampled fish (Hence there was no indication of toxic effect of the test chemical, thereby fulfilling the validity criteria and hence, study was considered to be valid.The Bio-concentration factor (BCF) was determined as 0,8394 and 0.4624 for 0.1 mg/L and 1.0 mg/L concentrations respectively in the uptake phase. Further, based on the overall rate constants of the uptake and depuration phases, the kinetic Bio-concentration factor (BCFk) was determined to be -0.1261 and -0.3114 for the concentrations 0.1 mg/L and 1.0 mg/L respectively. From the growth rate analysis, the growth corrected kinetic bio-concentration factor (BCFkg) was determined to be -0.1240 and -0.3173 for the concentrations 0.1 mg/L and 1.0 mg/L respectively. Based on the lipid content analysis, the lipid normalized kinetic bio-concentration factor (BCFkl) was determined to be -0.1852 and -0.5097 for the concentrations 0.1 mg/L and 1.0 mg/L respectively in the uptake phase. Since, the BCF value did not exceeded the threshold limit of 2000, thus, test chemical was considered to be non-bioaccumulative,