Benzene-1,4-diammonium sulphate

Endpoint summary

Description of key information

Short term toxicity to fish:

Study was conducted to access the effect of test chemical Benzene-1,4-diammonium sulphate on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 1g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous 48 hours stirring for achieving test concentrations of 0.3125mg/L, 0.625mg/L,1.25mg/L, 2.5mg/L,5mg/L respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test chemical Benzene-1,4-diammonium sulphate on Zebra Fish Danio rerio was determine to be > 2.5 to < 5 mg/l. Test consider to be valid as it fulfills all necessary criteria as per the standards. Based on the LC50 value, it can be consider that the chemical Benzene-1,4-diammonium sulphate was toxic and can be consider to be classified as aquatic chronic 2 category as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrates:

Aim of this study was to assess the short term toxicity of Benzene-1,4-diammonium sulphate to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs. Short term toxicity of test chemical Benzene-1,4-diammonium sulphate to aquatic invertebrates was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system. 0, 0.05, 0.10, 0.20, 0.40, 0.80 mg/l nominal concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance Benzene-1,4-diammonium sulphate, in Daphnia magna was determined to be 0.37 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, it was concluded that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic acute 1 category as per the CLP classification criteria.

 

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of test chemical Benzene-1,4-diammonium sulphate when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD guideline 201. The stock solution 10 mg/l was prepared by dissolving white powder in OECD growth medium. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Various concentrations 0, 0.31, 0.62, 1.24, 2.50, 5 and 10 mg/l were used. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration ErC50 was calculated using nonlinear regression by the software Prism 4.0. Effect on the growth of algae was determine after an exposure period of 72 hrs. Based on the growth rate inhibition of algae Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) due to the exposure of chemical Benzene-1,4-diammonium sulphate the ErC50 was determine to be 3.49 mg/l. Based on the ErC50 value, the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 2 category as per the CLP criteria.

 

Toxicity to microorganisms:

1. Based on the population growth inhibition of Tetrahymena pyriformis by the test chemical after 60 hrs, the IGC50 was observed at 10.38 mg/l.

2. Based on the population growth inhibition of Tetrahymena pyriformis by the test chemical after 60 hrs, the IGC50 was observed at 7.35 mg/l.

3. As the two different species Ruminococcus albus and Butyrivibrio fibrisolvens exposed with the test chemical, effect on the growth of organisms vary. The IGC for Ruminococcus albus was determine to be 10 mg/l and on the other species it was observed that the growth inhibited at 100 mg/l.

Thus based on the overall studies for Benzene-1,4-diammonium sulphate(CAS No. 16245 -77 -5), toxicity value ranges from 10 mg/l to 10.38 mg/l.

Additional information

Summarized result of toxicity of the chemical Benzene-1,4-diammonium sulphate (CAS No. 16245-77-5)on the growth of fishes, aquatic invertebrates and algae and microorganisms was studied by considering and collecting the data from various databases for target chemical. The studies are as follows:  

 

Short term toxicity to fish:

Study was conducted to access the effect of test chemical Benzene-1,4-diammonium sulphate on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 1g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous 48 hours stirring for achieving test concentrations of 0.3125mg/L, 0.625mg/L,1.25mg/L, 2.5mg/L,5mg/L respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test chemical Benzene-1,4-diammonium sulphate on Zebra Fish Danio rerio was determine to be > 2.5 to < 5 mg/l. Test consider to be valid as it fulfills all necessary criteria as per the standards. Based on the LC50 value, it can be consider that the chemical Benzene-1,4-diammonium sulphate was toxic and can be consider to be classified as aquatic chronic 2 category as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrates:

Based on the experimental data from various sources for the target chemical and read across chemical, study have been reviewed to determine the mode of action of Benzene-1,4-diammonium sulphate on the mobility rate and behavior of aquatic invertebrates. The studies are as mentioned below:

 

In the first key study from experimental report 2017. Aim of this study was to assess the short term toxicity of Benzene-1,4-diammonium sulphate to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs. Short term toxicity of test chemical Benzene-1,4-diammonium sulphate to aquatic invertebrates was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system. 0, 0.05, 0.10, 0.20, 0.40, 0.80 mg/l nominal concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance Benzene-1,4-diammonium sulphate, in Daphnia magna was determined to be 0.37 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, it was concluded that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic acute 1 category as per the CLP classification criteria.

 

First study was supported by the second experimental study 1985 for structurally similar read across chemical. Equal volumes of laboratory mass culture well water were collected from all Daphnia mass culture tanks, pooled, and filtered for use as dilution water.  The test material was prepared as a 0.1 mg/mL stock solution in filtered laboratory mass culture well water, and diluted with filtered laboratory mass culture well water in 250 mL glass test vessels to yield the desired nominal concentrations in 200 mL final volumes. 250 mL glass beakers, 200 mL volume. The substance was tested on Daphnia magna according OECD guideline 202 under static unaerated test conditions during a 48-hour exposure period. At the beginning of the test, the measured concentrations of the test substance ranged from 11-60% of the nominal concentrations. Because the measured concentrations were near or below the limit of detection at the beginning of the test, samples taken at the end of the 48-hour exposure period were not analyzed. After the exposure of test chemical for 48-h immobility were observed. The EC50 in Daphnia magna was observed at 0.25 mg/L (nominal concentration).

 

Similarly in the third study from report 2002, Study was conducted to determine the toxicity of test chemical on the growth of aquatic invertebrates. Test conducted accordance with OECD guideline. An acute, semi-static test was conducted at concentrations of 0, 0.12, 0.18, 0.26, 0.38, 0.54, 0.82, and 1.22 mg/L (measured) on Daphnia magna. Based on the results of a preliminary test, showing a 100% daphnia immobilisation rate in the 1.8 mg/L concentration plot and no inhibition at all at a concentration of less than 0.22 mg/L, for this test seven concentration plots of 2.2 mg/L or less were prepared.  The test plots were as follows:  Control and 0.22, 0.32, 0.46, 0.68, 1.0, 1.5 and 2.2 mg/L. Test conducted in 4 replicates. Exposure after 48 hours resulted in 0, 0, 5, 35, 45, 100, 100 and 100% immobility, respectively. Based on the measured concentration the EC50 was 0.33 mg/L.

 

In the fourth supporting study 1992, A Daphnia study was performed under static-renewal and OECD 202 guideline. During this study the test organisms were intentionally fed to help assess the potential effects of feeding. At the time of analysis the sample was thawed and pipetted into a syringe and filtered through a filter disk into a disposable culture tube.  Test solutions were diluted to nominal concentrations of 10 µg/L.  Those concentrations which were nominally less than or equal to 10 µg/L were prepared without further dilution. An aliquot of each solution and 10x sample buffer were mixed in a disposable culture tube.  Fluorescamine was added to the sample mixture and vortexed repeatedly to allow for turbulent mixing.  The resulting solution was transferred into an amber auto sampler vial and capped. A stock solution was prepared in dilution water and stirred/sonicated for 20 minutes.  Aliquots of stock solution were immediately added to the dilution water to create 500 mL of test solution.  A graduated cylinder was used to distribute 200 mL of test solution to each replicate beaker. Nominal concentrations were 0.010, 0.026, 0.075, 0.21, 0.60, 1.74, and 5.0 mg/L. Mean measured concentrations 24 hours after test solution preparation were 0.0045, 0.013, 0.051, 0.092, 0.16, 0.21, and 0.57 mg/L, respectively. The 48-hour EC50, based on arithmetic mean, measured concentrations and immobility, was 0.15 mg/L with a 95% confidence interval of 0.13-0.17 mg/L. 

 

In the fifth study the substance was tested on Daphnia magna according OECD guideline 202 under static unaerated test conditions during a 48-hour exposure period. Chemical analytically monitorized. Equal volumes of laboratory mass culture well water were collected from all Daphnia mass culture tanks, pooled, and filtered for use as dilution water.  The test material was prepared as a 0.1 mg/mL stock solution in filtered laboratory mass culture well water, and diluted with filtered laboratory mass culture well water to yield the desired nominal concentrations. After mixing, 200 mL aliquots of each concentration were introduced into each of two separate 250 mL glass test vessels. At the beginning of the test, the measured concentrations of the test substance ranged from below detection up to 50% of the nominal concentrations. Because the measured concentrations were near or below the limit of detection at the beginning of the test, samples taken at the end of the 48-hour exposure period were not analyzed. Based on the nominal concentrations, the 48-hour EC50 was 0.28 mg/L, with a 95% confidence interval of 0.24 to 0.33 mg/L.

 

Sixth study report for read across chemical also supports the classification of test chemical. The acute toxicity of the test substance to Daphnia magna was investigated in a study according to OECD TG 202 and GLP (DR. U. NOACK LABORATORIEN, 2013a). All concentration levels of the test item and the control were analytically verified in the fresh media (0 and 24 hours) and old media (24 and 48 hours). Directly after preparation of the test solutions, the fresh media samples were taken in duplicate. Date and time of preparation of the test solutions and use in the test were noted. For each sample, 10 mL of the test solution was diluted for stabilization with 10 mL HPLC water (containing 20% acetonitrile, 0.2% TFA and 20% of a 0.2% ascorbic acid solution) and analysed the day of collection within 4 hours of sampling. For the old media, samples were taken directly from the test vessels containing daphnids. The old media samples were prepared as described for the fresh media samples. In this study daphnids (4 replicates of 5 daphnids per concentration) were exposed to nominal test substance concentrations of 0, 45, 100, 220, 484, 1065 and 2343 µg/L for 48 hours under semi static conditions. Analysis of test concentrations showed that recovery rates of test item in the fresh media (0 and 24 hours) were in the range of 96 to 105 % of the nominal values at all tested concentration levels. The recovery rates in the old media (24 and 48 hours) were in the range of 26 to 73 % of the nominal values. The geometric mean recovery rates were in the range of 50 to 82 %. The geometric mean measured concentrations of test item were calculated to be 31.2, 78.4, 181, 326, 628 and 1176 µg/L. After 48 hours exposure, immobilisation was noted first at a measured concentration of 0.326 mg/L (i.e. 25% of immobilisation, mean value). At the highest test concentration 100% immobilisation was observed. Based on the immobilisation rates the EC50 value was calculated to be 0.496 mg/L (496 µg/L) (based on geometric mean measured concentration).

 

Thus based on the overall studies, it was concluded that the chemical was toxic to the growth of aquatic invertebrates, and classified as aquatic acute 1 as per the CLP classification criteria.

 

 

 

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of test chemical Benzene-1,4-diammonium sulphate when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD guideline 201. The stock solution 10 mg/l was prepared by dissolving white powder in OECD growth medium. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Various concentrations 0, 0.31, 0.62, 1.24, 2.50, 5 and 10 mg/l were used. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration ErC50 was calculated using nonlinear regression by the software Prism 4.0. Effect on the growth of algae was determine after an exposure period of 72 hrs. Based on the growth rate inhibition of algae Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) due to the exposure of chemical Benzene-1,4-diammonium sulphate the ErC50 was determine to be 3.49 mg/l. Based on the ErC50 value, the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 2 category as per the CLP criteria.

 

Toxicity to microorganisms:

Toxicity of Benzene-1,4-diammonium sulphate (CAS No. 16245 -77 -5) on the growth and other biological activity of microorganisms is predicted on the basis of it structurally and functionally similar read across chemicals. The studies are as mentioned below:

 

In the first study toxicity was measured. Study was conducted to investigate the effect of test chemical on the growth of ciliate Tetrahymena pyriformis. Test conducted under the static system for 60 hrs. Chemical analytically monetarized by using HPLC. DMSO used as a solvent. Axenic cultures of T. pyriformis reared in a semidefined protease peptone medium were grown at 28°C under a static regime. 250 ml Erlenmeyer flask containing 50 ml of test solution. Duplicate assembly were arrange for the test. Tetrahymena pyriformis was used as the bioassay system. Population growth and density being selected on which toxicity were measured. Based on the population growth inhibition of Tetrahymena pyriformis by the test chemical after 60 hrs, the IGC50 was observed at 10.38 mg/l.

 

First study was supported by the second experimental study from peer reviewed journal. Aim of this study was to investigate the effect of test chemical on the growth of ciliate Tetrahymena pyriformis. Test conducted under the static system for 60 hrs. Chemical analytically monetarized by using HPLC. DMSO used as a solvent. Axenic cultures of T. pyriformis reared in a semidefined protease peptone medium were grown at 28°C under a static regime. 250 ml Erlenmeyer flask containing 50 ml of test solution. Duplicate assembly were arrange for the test. Tetrahymena pyriformis was used as the bioassay system. Population growth and density being selected on which toxicity were measured. Based on the population growth inhibition of Tetrahymena pyriformis by the test chemical after 60 hrs, the IGC50 was observed at 7.35 mg/l with 95% CI was 5.95 – 8.98 mg/l.

 

In third study toxicity were observed at 10 mg/l. Aim of this study was to determine the toxicity of test chemical on the growth of microorganisms. Test conducted on two different organisms Ruminococcus albus and Butyrivibrio fibrisolvens. As the two different species Ruminococcus albus and Butyrivibrio fibrisolvens exposed with the test chemical, effect on the growth of organisms vary. The IGC for Ruminococcus albus was determine to be 10 mg/l and on the other species it was observed that the growth inhibited at 100 mg/l.

 

Thus based on the overall studies for Benzene-1,4-diammonium sulphate(CAS No. 16245 -77 -5), toxicity value ranges from 10 mg/l to 10.38 mg/l.

Based on the overall studies chemical consider to be toxic and classified as aquatic acute 1 as per the CLP classification criteria.