Benzene-1,4-diammonium sulphate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17.07.2017 to 19.07.2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed using standard test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

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.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (IUPAC name): Benzene-1,4-diammonium sulphate- Molecular formula: C6H8N2.H2O4S- Molecular weight: 206.221 g/mol- Smiles notation: S(=O)(=O)(O)O.c1(ccc(cc1)N)N- InChI: 1S/C6H8N2.H2O4S/c7-5-1-2-6(8)4-3-5;1-5(2,3)4/h1-4H,7-8H2;(H2,1,2,3,4)- Substance type: Organic - Physical state: Solid

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

The stock solution 10 mg/l was prepared by dissolving white powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM- Common name: Water flea- Strain: Straus- Source: Own breeding at University of Chemistry and Technology, Prague- Age at study initiation (mean and range, SD): The animals used for the test shall be less than 24 h old and should not be first brood progeny- Feeding during test: No feedingACCLIMATION - No data available- Acclimation period:- Acclimation conditions (same as test or not):- Type and amount of food:- Feeding frequency:- Health during acclimation (any mortality observed):

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Remarks on exposure duration:

± 1 hr

Test temperature:

20±1°C

pH:

pH at concentration 0.80 mg/l: 8 changes to 7.9 during testControl: 8 did not change during test

Dissolved oxygen:

higher than 7.4 mg/L at the end of test

Nominal and measured concentrations:

0, 0.05, 0.10, 0.20, 0.40, 0.80 mg/l

Details on test conditions:

TEST SYSTEM- Test vessel: 50 ml glass vessel- fill volume: 25 ml- No. of organisms per vessel: 5- No. of vessels per concentration (replicates): 4TEST MEDIUM / WATER PARAMETERS- Source/preparation of dilution water: Natural water (surface or ground water), reconstituted water or dechlorinated tap water are acceptable as culturing and dilution water if D. magna survives in it for the duration of the culturing, acclimation and testing without showing signs of stress. Waters in the range pH 6 to pH 9, with hardness between 140 mg/l and 275 mg/l (as CaCO3) are recommended.As an example, the preparation of dilution water meeting the requirements is described below.Dissolve known quantities of reagents in water. The dilution water prepared shall have a pH of 7.8 ± 0.5, a hardness of (225 ± 50) mg/l (expressed as CaCO3), a molar Ca + Mg ratio close to 4 + 1 and a dissolved oxygen concentration above 7 mg/l.Prepare the solutions specified below:- Calcium chloride solution: Dissolve 117.6 g of calcium chloride dihydrate (CaCl2.2H2O) in water (4.2) and make up to 1 l with water (4.2).- Magnesium sulfate solution: Dissolve 49.3 g of magnesium sulfate heptahydrate (MgSO4.7H2O) in water (4.2) and make up to 1 l with water (4.2).- Sodium bicarbonate solution: Dissolve 25.9 g of sodium bicarbonate (NaHCO3) in water (4.2) and make up to 1 l with water (4.2).- Potassium chloride solution: Dissolve 2.3 g of potassium chloride (KCI) in water (4.2) and make up to 1 l with water (4.2).MixingMix 2.5 ml of each of the four solutions and make up to 1 l with water.The dilution water shall be aerated until the dissolved oxygen concentration has reached saturation and the pH has stabilized. If necessary, adjust the pH to 7.8 ± 0.5 by adding sodium hydroxide (NaOH) solution or hydrochloric acid (HCI). The dilution water prepared in this way shall not be further aerated before use.- Sodium hydroxide solution, e.g. [NaOH] : 1 mol/l.- Hydrochloric acid, e.g. [HCl] : 1 mol/l.Reference substance: Dissolve 600 mg of potassium dichromate (K2Cr2O7) in water and make up to 1 l with water (4.2).OTHER TEST CONDITIONS- Adjustment of pH: no adjustment done- Photoperiod: No - Darkness- Light intensity:CALCULATION:EC50 was calculated using non linear regression by the software Prism 4.0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

0.37 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI was 0.30 - 0.47 mg/l

Results with reference substance (positive control):

- Results with reference substance valid- EC50: 0.73 mg/L (24 hours)

Reported statistics and error estimates:

EC50 was calculated using non linear regression by the software Prism 4.0

Validity criteria fulfilled:

yes

Conclusions:

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.

Executive summary:

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.

Description of key information

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.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.37 mg/L

Additional information

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.