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EC number: 210-431-8 | CAS number: 615-50-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Toxicity to aquatic algae and cyanobacteria
Administrative data
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From Oct. 15, 2012 to Oct. 25, 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Study well documented, followed guideline, GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 012
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- according to OECD and German principles of GLP
Test material
- Reference substance name:
- 2-methyl-p-phenylenediamine sulfate
- EC Number:
- 210-431-8
- EC Name:
- 2-methyl-p-phenylenediamine sulfate
- Cas Number:
- 615-50-9
- Molecular formula:
- C7H10N2.H2O4S
- IUPAC Name:
- 2-methylbenzene-1,4-diamine; sulfuric acid
- Reference substance name:
- toluene-2,5-diamine sulfate
- IUPAC Name:
- toluene-2,5-diamine sulfate
- Test material form:
- solid: particulate/powder
- Remarks:
- migrated information: powder
- Details on test material:
- - Name of test material: Toluene-2,5-diamine sulfate (Code: 95353654)
- TSIN: Not reported
- Substance type: Pure active substance
- Physical state: White to light pink or light grey powder
- Stability under test conditions: Not reported
- Storage condition of test material: Room temperature, protected from moisture and light
- Water solubility: 5.03 g/L
Constituent 1
Constituent 2
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 0, 1.0, 2.20, 4.84, 10.6 and 23.4 mg/L
- Sampling period: The concentrations of test material were analysed at the beginning (0 hours) and after 6, 12, 18, 24, 48 and 72 hours in all concentration levels of the test material and the control.
- Sampling method: Sampling was performed in the following manner:
(i) At 0 hour: For the fresh media, samples of the freshly prepared test solutions (without algae) were taken and analysed.
(ii) After 6, 12 and 18 hours: Samples were taken from additionally prepared replicates (with algae) per concentration level and control.
(iii) After 24, 48 and 72 hours: Samples were taken directly from the test replicates.
1 mL per replicate was pooled for each treatment and from this pool 2 x 0.8 mL samples were taken for analysis.
- Sample storage: Retention samples for possible re-analyses were stored at room temperature.
Test solutions
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 32 mg/L of stock solution was freshly prepared with dilution water. The dilution water was prepared in accordance with OECD guideline 201.
- Dispersion method: The stock solution was treated with ultrasound for 2 min at room temperature.
- Application of test solution: Test solution application was carried out by adding appropriate volumes of the stock solution to the test replicates.
- Controls: Prepared using dilution water without test material
Test organisms
- Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- TEST ORGANISM
- Common name: Green alga
- Strain: CHODAT SAG 86.81
- Source: Sammlung von Algenkulturen (SAG), Pflanzenphysiologisches Institut der Universitat Gottingen, Nikolausberger Weg 18, 0-37073 Gottingen
- Age of inoculum: 3 -4 days old pre culture
- Method of cultivation: Fresh stocks were prepared every month on Z-Agar. Light intensity amounted 35-70 µE. m (-2). s(-1) for 24 h/d. Nutrient medium Z according to LOTTGE et al. (1994) was used as culture medium.
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- 4 - 7 days (recovery phase in fresh untreated test medium after 72 h of exposure period)
Test conditions
- Hardness:
- 0.24 mmol Ca+Mg/L (nominal hardness)
- Test temperature:
- 21 - 24°C, controlled at ± 2 °C (actual: 22-23°C, mean value: 22.5°C; measured by hygrothermograph)
Temperature measured continuously was determined to be within the acceptable limits. - pH:
- 7.91-8.21 (pH range from 0 to 72 hours; measured by pH-Meter)
pH values were determined to be within the acceptable limits. - Nominal and measured concentrations:
- Nominal concentration: 0, 1, 2.2, 4.84, 10.6 and 23.4 mg/L
Time weighed average measured concentrations (0 – 72 hours): 0, 0.14, 0.31, 0.62, 1.25 and 2.47 mg/L. The percent of nominal of test material were in the range of 98 to 109 % at test start and decreased as expected to the range of < LOQM to 1 % at the test end of the exposure. - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Type: Closed (sealed with cotton wool plugs)
- Material, fill volume: Sterile 250 mL Erlenmeyer flasks containing 100 mL of test solution
- Aeration: No
- Agitation: Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm.
- Initial cells density: 4332 cells/mL (mean of replicates)
- Control end cells density: 877721 cells/mL (mean of replicates)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: Yes, OECD media.
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was prepared in accordance with standard OECD guideline 201. Detailed composition is provided in the study report.
- Ca/Mg ratio: CaCl2.2H2O: 18 mg/L/ (∑MgCl2.6H2O: 12 mg/L,MgSO4.7H2O: 15 mg/L)
- Hardness: 0.24 mmol Ca+Mg/L
No information on any other water parameters was provided in the study report.
- Culture medium different from test medium: Yes, Nutrient medium Z according to LUTTGE et al. (1994) was used as the culture medium.
- Intervals of water quality measurement: The pH-value at the beginning of the test was measured from one additional replicate of each concentration and control. At the end it was measured from pooled replicates per concentration and control. The room temperature was measured continuously by a hygrothermograph.
OTHER TEST CONDITIONS
- Sterile test conditions: Yes
- Adjustment of pH: No
- Photoperiod: 24 hours/day light
- Light intensity and quality: 60 - 120 µE/m(2)/sec
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell density: The cell density was measured at the beginning of the test (after application) and every 24 hours via Chlorophyll-a- fluorescence, excitation at 436 nm, emission at 685 nm. Dilution water with test material at the appropriate concentration but without algae was used as background signal
- Microscopic evaluation: Microscopic evaluation of cells was carried out at the start and end of the incubation. The cells were checked for unusual cell shapes, color differences, differences in chloroplast morphology, flocculation, adherence of algae to test containers and agglutination of algae cells.
TEST CONCENTRATIONS
- Spacing factor for nominal test concentrations: 2.2
- Range finding study: Yes. A non-GLP range finding study was performed out at 0.1, 1, 10 and 100 mg/L (nominal) concentrations (with batch R0019281).
- Results used to determine the conditions for the definitive study: Growth rate inhibition: 100%; Yield inhibition: 100% at 100 mg/L (highest concentration tested). Based on the results of range finding study 5 concentrations were selected for main test.
Refer to ‘Table 2’ under ‘Any other information on results incl. tables’ section below. - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.653 mg/L
- Nominal / measured:
- meas. (not specified)
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 0.61 – 0.7 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.31 mg/L
- Nominal / measured:
- meas. (not specified)
- Conc. based on:
- act. ingr.
- Basis for effect:
- other: inhibition of yield
- Remarks on result:
- other: 0.29 – 0.33 mg/L
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 7.5 - 8.4
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 4.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 4.0 - 5.1
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 4 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- other: inhibition of yield
- Remarks on result:
- other: 3.7 - 4.2
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 2.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- other: inhibition of yield
- Remarks on result:
- other: 2.0 - 2.5
- Details on results:
- For details on cell density, see ‘Table 3’ below in the ‘Any other information on results incl. tables’ section.
- Exponential growth in the control: Yes, after 72 hours of exposure, mean cell density in control group was 877721 cells/mL. This was an increase of 203 times over the initial inoculation density.
- Microscopic evaluation: Microscopic evaluation of the cells at start and end of the incubation period revealed no morphological abnormalities.
- Any stimulation of growth found in any treatment: No, for details on growth rate, see 'Table 4' below in the ‘Any other information on results incl. tables’ section.
- Effect concentrations exceeding solubility of substance in test medium: No - Results with reference substance (positive control):
- - Results with reference substance valid: Yes, the results of positive control were in the valid range.
- EyC50 (Yield): 0.324 mg/L; 95% CI: 0.304-0.347 (Valid range: 0.319 ± 0.140)
- ErC50 (Growth rate): 0.54 mg/L; 95% CI: 0.528-0.551 (Valid range: 0.686 ± 0.470) - Reported statistics and error estimates:
- EC10, EC20 and EC50 values with confidence intervals of growth rate and yield inhibition after 72 hours were calculated via non linear regression according to BRUCE & VERSTEEG (1992).
Any other information on results incl. tables
Recovery of algal cells
After 72hours algae were transferred from the three highest concentrations (time weighed average measured concentrations: 0.62 – 2.47 mg/L) and the control to fresh untreated medium and allowed to grow for further 4 – 7 days under test conditions.
The test material effect was observed to be reversible at these concentrations. Therefore, there is potential for recovery following exposure up to 2.47 mg/L (highest test concentration).
Table 2: Results of range finding test (0 - 72 hours)
Nominal Test material Concentration |
Growth Rate Inhibition |
Yield Inhibition |
100 |
100 |
100 |
10.0 |
51 |
94 |
1.00 |
- 9 |
- 60 |
0.1 |
- 7 |
- 41 |
2 replicates per concentration level, 4 replicates for the control
Table 3: Effect of Toluene-2,5-Diamine Sulfate on Cell densities of green algae (Desmodesmus subspicatus) during a 72 hours exposure (Study # 66353)
Time weighed average measured concentrations [mg/L] |
Replicate No. |
Cell density [cells/mL] |
|||
0 hour |
24 hours |
48 hours |
72 hours |
||
2.47 |
1 |
4332 |
< LOQ |
< LOQ |
2753 |
2 |
4332 |
3621 |
< LOQ |
2800 |
|
3 |
4332 |
< LOQ |
< LOQ |
2734 |
|
Mean |
4332 |
n.a. |
n.a. |
2762 |
|
1.25 |
1 |
4332 |
2916 |
8187 |
15930 |
2 |
4332 |
4489 |
10357 |
20380 |
|
3 |
4332 |
< LOQ |
10417 |
21837 |
|
Mean |
4332 |
n.a. |
9654 |
19382 |
|
0.62 |
1 |
4332 |
11851 |
79164 |
301075 |
2 |
4332 |
11405 |
56995 |
273814 |
|
3 |
4332 |
11512 |
68049 |
297464 |
|
Mean |
4332 |
11589 |
68069 |
290784 |
|
0.31 |
1 |
4332 |
14848 |
147938 |
764500 |
2 |
4332 |
17352 |
151411 |
899137 |
|
3 |
4332 |
16660 |
156298 |
893885 |
|
Mean |
4332 |
16287 |
151882 |
852507 |
|
0.14 |
1 |
4332 |
24378 |
182483 |
994808 |
2 |
4332 |
21399 |
169519 |
970875 |
|
3 |
4332 |
16880 |
170400 |
920932 |
|
Mean |
4332 |
20886 |
174134 |
962205 |
|
Control |
1 |
4332 |
22937 |
160474 |
850239 |
2 |
4332 |
28498 |
171249 |
952766 |
|
3 |
4332 |
20494 |
164343 |
884016 |
|
4 |
4332 |
26287 |
179433 |
927889 |
|
5 |
4332 |
25082 |
171859 |
801869 |
|
6 |
4332 |
22299 |
159110 |
849547 |
|
Mean |
4332 |
24266 |
167745 |
877721 |
n.a. = Not Applicable
LOQ = Limit of quantification (2280 cells/mL)
Table 4: Inhibition of yield and growth rate evaluation after 72 hours of exposure (Study # 66353)
Time weighed average measured concentrations |
Replicate |
Growth rate |
Growth rate inhibition |
Yield |
Inhibition of yield |
||
[mg/L] |
No. |
[d-1] |
[%] |
[cells/mL] |
[%] |
||
2.47 |
1 |
|
-0.151 |
100 |
|
-1579 |
100 |
2 |
|
-0.145 |
100 |
|
-1532 |
100 |
|
3 |
|
-0.153 |
100 |
|
-1598 |
100 |
|
Mean |
(+) |
-0.150 |
100 |
(+) |
-1570 |
100 |
|
1.25 |
1 |
|
0.434 |
75.5 |
|
11598 |
98.7 |
2 |
|
0.516 |
70.8 |
|
16048 |
98.2 |
|
3 |
|
0.539 |
69.5 |
|
17505 |
98.0 |
|
Mean |
(+) |
0.496 |
72.0 |
(+) |
15050 |
98.3 |
|
0.62 |
1 |
|
1.41 |
20.1 |
|
296743 |
66.0 |
2 |
|
1.38 |
21.9 |
|
269482 |
69.2 |
|
3 |
|
1.41 |
20.4 |
|
293132 |
66.4 |
|
Mean |
(+) |
1.40 |
20.8 |
(+) |
286452 |
67.2 |
|
0.31 |
1 |
|
1.72 |
2.57 |
|
760168 |
13.0 |
2 |
|
1.78 |
-0.49 |
|
894805 |
-2.45 |
|
3 |
|
1.78 |
-0.38 |
|
889553 |
-1.85 |
|
Mean |
(-) |
1.76 |
0.57 |
(-) |
848175 |
2.89 |
|
0.14 |
1 |
|
1.81 |
-2.39 |
|
990476 |
-13.4 |
2 |
|
1.80 |
-1.93 |
|
966543 |
-10.7 |
|
3 |
|
1.79 |
-0.94 |
|
916600 |
-4.95 |
|
Mean |
(-) |
1.80 |
-1.75 |
(-) |
957873 |
-9.68 |
|
Control |
1 |
|
1.76 |
|
|
845907 |
|
2 |
|
1.80 |
|
|
948434 |
|
|
3 |
|
1.77 |
|
|
879684 |
|
|
4 |
|
1.79 |
|
|
923557 |
|
|
5 |
|
1.74 |
|
|
797537 |
|
|
6 |
|
1.76 |
|
|
845215 |
|
|
Mean |
|
1.77 |
|
|
873389 |
|
Negative inhibitions = Increase of growth
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- The 72 h EC50 value of Toluene-2,5-Diamine sulphate to Desmodesmus subspicatus for inhibition of growth rate and inhibition of yield were 0.653 and 0.31 mg/L respectively, based on geometric mean measured concentrations, and 8.0 and 4.0 mg/l based on nominal concentrations.
- Executive summary:
The toxicity of Toluene-2,5-Diamine sulphate to Desmodesmus subspicatus was determined in a 72 hours algae growth inhibition test, following OECD 201 guideline under static conditions.
The nominal test concentrations were 0 (control), 1.0, 2.20, 4.84, 10.6 and 23.4 mg/L. Time weighed average measured concentrations were 0, 0.14, 0.31, 0.62, 1.25 and 2.47 mg/L.The percent of nominal of test material were in the range of 98 to 109 % at test start and decreased as expected to the range of < LOQM to 1 % at the test end of the exposure.
Endpoints evaluated were inhibition of growth rate and inhibition of yield over a period of 72 hours. 3 replicates was tested for each test concentration and 6 replicates for control.
The test material was found to inhibit the growth of freshwater green alga after 72 hours of exposure with the following effects :
1. Based on geometric mean measured concentrations):
EyC50: 0.31 mg/L (based on inhibition of yield),
ErC50: 0.653 mg/L (based on inhibition of growth rate)
2. Based on nominal concentrations:
EyC50: 4.0 mg/L (based on inhibition of yield),
ErC50: 8.0 mg/L (based on inhibition of growth rate)
After 72 hours algae were transferred from the three highest concentrations (time weighed average measured: 0.62 – 2.47 mg/L) and the control to fresh untreated medium and allowed to grow for further 4 – 7 days under test conditions. The test material effect was observed to be reversible at all test concentrations following 72 h exposure. Therefore, there is potential for recovery following exposure up to 2.47 mg/L (highest test concentration).
This toxicity test is classified as acceptable and satisfies the guideline requirements for the OECD Guideline 201.
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