Hydrogen tetrasodium bis[2-[[6-[[4-chloro-6-[3-sulphoanilino]-1,3,5-triazin-2-yl]amino]-1-hydroxy-3-sulpho-2-naphthyl]azo]benzoato(4-)]chromate(5-)

Endpoint summary

Description of key information

Short term toxicity to aquatic invertebrates

On the basis of the experimental studies of the read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism Daphnia magna, the 48hr EC50 value can be expected to be > 50 mg/l. Since the test chemical is readily biodegradable in nature, test chemical was considered as non-toxic and hence, considered to be not classified as per CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria

On the basis of the experimental studies of the read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism, the 96 hr LC50 value can be expected to be > 65 mg/l. Since the test chemical is readily biodegradable in nature, test chemical was considered as non-toxic and hence, considered to be not classified as per CLP classification criteria.

Additional information

Short term toxicity to aquatic invertebrates

Data available of the structurally and functionally similar read across chemicals has been reviewed to determine the effect of the test chemical on aquatic invertebrates. The studies are as mentioned below:

 

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on Daphnia magna. The test was performed in accordance to OECD guideline No. 202“Daphnia sp.,Acute Immobilization Test”. The test solution was prepared by dissolving 50 mg of the test chemical in 100 ml of ADaM’s media achieving a final test concentrations of 500 mg/l. Test chemical concentrations used for the study were 0, 7.5, 15, 30, 60, 120 mg/l, respectively. Test concentrations were verified analytically by UV-Vis Spectrophotometer. Study was performed using 10 daphnids in a static system. Total 10 Daphnids/conc. were exposed to test chemical in 25 ml beakers in a volume of 20 ml of liquid solution containing both the chemical and media. The beakers were placed in a room at a temperature of 18 -22°C, hardness of water 140 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions with light intensity 1000 – 1500 Lux, respectively. One control vessel was also run simultaneously during the study. The animals were exposed to medium (i.e.a beaker containing only medium) and the test chemical concentrations for a period of 48 hour. Potassium dichromate was used as a reference substance for the study. The 48 hr EC50 value of reference substance was determined to be 0.831 mg/l. No Immobility were found in the control test animals and the dissolved oxygen concentration at the end of the test in the control and test vessel was ≥ 3 mg/l, thus validity criterion of the study has been fulfilled. As the concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. On the basis of effect of test chemical on mobility of the test organism, the median effect concentration (EC50 (48 h)) value was determined to be > 120 mg/L. Thus, based on the EC50 value, chemical was considered as non-toxic to aquatic invertebrates and hence, considered to be 'not classified' as per CLP classification criteria.

 

Another short term toxicity to aq. Invertebrate study was conducted for 48 hrs for assessing the effect of test chemical. The study was performed in accordance to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system. Daphnia magna was used as a test organism for the study.On the basis of effect on the mobility of the test organism Daphnia magna, the 48 hr EC50 value was determined to be > 50 mg/l. Since the test chemical is readily biodegradable in water, chemical was considered as non-toxic to aquatic invertebrates and hence, considered to be not classified as per the CLP classification criteria.

 

On the basis of the experimental studies of the read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism Daphnia magna, the 48hr EC50 value can be expected to be > 50 mg/l. Since the test chemical is readily biodegradable in nature, test chemical was considered as non-toxic and hence, considered to be not classified as per CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria

Data available of the structurally and functionally similar read across chemicals has been reviewed to determine the effect of the test chemical on aquatic fishes. The studies are as mentioned below:

 

A freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. Sterile, unicellular, suspension cultures of algae Pseudokirchneriella subcapitata of length 8 – 14 μm and width 2 - 3 μm was used as a test organism. OECD medium composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test solution was prepared by dissolving 200 mg of test chemical in 200 ml of OECD medium to get the final concentration of 1000 mg/L. Test chemical concentrations were verified analytically by UV-VIS spectrophometer. Green algae were exposed to six different nominal concentrations of test chemical (0, 6.25, 12.5, 25, 50, 100 and 200 mg/L) in 100 ml conical flasks. Test vessel were placed in orbital shaking incubator for 72 hrs at a room at a temperature of 21 to 24 ± 2°C under a photoperiod of 16:8 hr light: dark conditions and with a continuous uniform illumination of 3000-4000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. Control containing medium without test chemical was used for the study. The cultures were counted and observed daily with the help of an automated cell counter to verify a normal and healthy appearance of the algae cells and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test chemical). Apart from this, the cell count of each test vessel was also noted with the help of a microscope. Cell densities were recorded in section by section growth rate at 24 hr intervals, which was calculated as specific growth rate. Potassium dichromate (K2Cr2O7) was used as a reference substance for the study. Since the concentration of the test chemical being tested has been satisfactorily maintained within ±20% of the nominal concentration throughout the test, all concentrations will be reported as nominal concentration. As per the OECD guideline No. 201 – Alga growth inhibition test, the biomass in the control cultures have increase exponentially by a factor of at least 16 within the 72 hr test period, the mean coefficient of variation by section specific growth rate in the control cultures not exceeded 35% and the coefficient of variation of average specific growth rate during the whole test period in replicate control cultures was not exceeded 10%, thus, fulfilling the validity of the criteria. The 72 hr EC50 value of reference substance was determined to be 0.809 mg/l. On the basis of growth rate of the test organism Pseudokirchneriella subcapitata, the 72 hrs median effect concentration (ErC50) was determined to be > 200 (nominal concentration). On the basis of the EC50 value, chemical was considered to be non-toxic to aquatic algae at environmental relevant concentrations and hence, considered to be not classified as per the CLP classification criteria.

 

Another toxicity to aquatic algae study was conducted for 72 hrs for assessing the effect of test chemical on green algae Pseudokirchneriella subcapitata. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. The study was performed using the same test procedure and under test conditions as mentioned in the above study. The test solution was prepared by dissolving 300 mg of test chemical in 300 ml of OECD Media This stock solution was kept for stirring for 48 hours to obtain a homogenous solution for the experiment. The stock solution was analyzed analytically by using double beam VWR spectrophotometer at day 0 and day 3, which has been satisfactorily maintained within ± 20 % of the nominal initial concentration throughout the test. Green algae were exposed to nominal concentration of test chemical (100 mg/l) in 100 ml conical flasks for 72 hrs. As per the OECD guideline No. 201 – Alga growth inhibition test, the biomass in the control cultures have increase exponentially by a factor of at least 16 within the 72 hr test period which corresponds to a specific growth rate of 0.92 per day, the mean coefficient of variation by section specific growth rate in the control cultures not exceeded 35% and the coefficient of variation of average specific growth rate during the whole test period in replicate control cultures was not exceeded 7%, thus, fulfilling the validity of the criteria. In control and test vessel, all cells appeared healthy, sickle shape and green throughout the study duration. On the basis of growth rate of the test organism Pseudokirchneriella subcapitata, the 72 hrs EC50 value was determined to be > 100 mg/l. Thus, based on the EC50 value, chemical can be considered as non-toxic to aquatic algae and thus can be considered to be not classified as per CLP classification criteria.

 

For the test chemical, toxicity to green algae study was carried out for assessing the effect of the test chemical. The study was conducted under static conditions for 72 hrs. On the basis of growth rate and AUG (biomass integral) obtained after exposure period of 72 hours, the median effect concentration (EC50) was determined to be > 65 and 20 mg/l & the NOEC value was evaluated to be 6.3 and 3.3 mg/l, respectively. Thus, based on the EC50 value, chemical can be considered as toxic to aquatic organisms. Since the test chemical is readily biodegradable in nature, test chemical was considered as non-toxic and hence, considered to be not classified as per CLP classification criteria.

 

On the basis of the experimental studies of the read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism, the 96 hr LC50 value can be expected to be > 65 mg/l. Since the test chemical is readily biodegradable in nature, test chemical was considered as non-toxic and hence, considered to be not classified as per CLP classification criteria.

On the basis of the available information of aquatic toxicity studies, it can be concluded that the test chemical can be considered as non-toxic to aquatic organisms at environmental relevant concentrations and considered to be ‘not classified’ as per CLP classification criteria.