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EC number: 224-923-5 | CAS number: 4553-62-2
- 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
Phototransformation in water
Administrative data
- Endpoint:
- phototransformation in water
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- August 2008
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: The study does not follow a particular guideline and is not GLP compliant. However, it is well described both in terms of experimental conditions and results, and allows to scientifically conclude on the results obtained during the algal study.
Cross-reference
- Reason / purpose for cross-reference:
- reference to other study
Reference
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- FROM 31 MARCH TO 23 AUGUST 2010
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: The study follows an internationaly recognized guideline (OECD 201) and is GLP compliant.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 10, 32, 100, 316, 1000 mg/L
- Sampling method: No data.
- Sample storage conditions before analysis: all samples were stored in a freezer (=< -10°C), protected from light until analysis was performed. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The highest test item concentration of 1000 mg test item/L was prepared by dissolving 500 mg test item into 500 mL test water by intense stirring for 15 minutes. The pH of this solution was adjusted with 0.1 M NaOH to pH 8.1. Adequate volumes of this solution were diluted with test water to prepare the test media of the other desired test concentations. The test media were prepared just before introduction of the algae.
- Controls: Test water without addition of test item.
- Blanks: One replicate of each test concentration and of the control was prepared without algae to provide a "blank" for the spectrophotometrical measurements. the additional replicates were incubated under the same conditions as the others.
No further data. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchnerella subcapitata
- Strain: 61.81 SAG
- Source (laboratory, culture collection): Sammlung von Algenkulturen, Pflanzenphysiologishces Institut der Universität Göttingen, 37073 Göttingen, Germany
- Age of inoculum (at test initiation): no data
- Method of cultivation: The algae were cultivated in the laboratories of IBACON under standardised conditions according to the test guidelines.
ACCLIMATION
- Acclimation period: The cells were taken from an exponentially growing pre-culture, which was set up 3 days prior to the test start under the same conditions as in the test.
- Culturing media and conditions (same as test or not): same as test
- Any deformed or abnormal cells observed: no - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Hardness:
- 0.24 mmol/L = 24 mg/L as CaCO3
- Test temperature:
- 22-23°C
- pH:
- from 8.0-8.1 at the beginning of the test, to 8.4-9.6 at the end of the test.
- Dissolved oxygen:
- no data
- Salinity:
- not applicable
- Nominal and measured concentrations:
- - nominal conc.: 0, 10, 32, 100, 316, and 1000 mg/L
- measured concentrations at 0h: 0, 9.17, 31.64, 104.90, 296.15, 1064.00 mg/L - Details on test conditions:
- TEST SYSTEM
- Test vessel: erlenmeyer flasks of 50 mL volume
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: erlenmeyer flasks of 50 mL volume filled with 50 mL of test medium
- Aeration: no data
- Initial cells density: 5000 cells/mL
- Control end cells density: 793970 cells/mL after 72 hours in control
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes (OECD medium)
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: analytical grade salts were added in deionised water.
- Total organic carbon: no data
- Particulate matter: no data
- Metals: no data
- Pesticides: no data
- Chlorine: no data
- Alkalinity: no data
- Ca/mg ratio: no data
- Conductivity < 5µScm-1
- Culture medium different from test medium: no
- Intervals of water quality measurement: The pH was measured in all test item concentrations and the control at the start and the end of the test. The appearance of the test item in test water was determined daily in all test item concentrations.
OTHER TEST CONDITIONS
- Sterile test conditions: no data
- Adjustment of pH: with 0.1 M NaOH in the stock solution
- Photoperiod: continuous (24:0)
- Light intensity and quality: 5892 lux
- Salinity (for marine algae): not applicable
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: The cell densities in the samples were determined by spectrophotometrical measurement. Therefore, defined volumes of the algal suspensions from all replicates and from the blanks were sampled after 24, 48 and 72 hours of exposure. The algal cell densities were calculated by substracting the absoprtion of the blanks, from each of the measured absorption of the test media. The cells densities in a number of samples from one control replicate were counted by microscope after 72 hours. Out of this control five defined dilutions were prepared and measured spectrophotometrically. Based on the counted cell densities and the absorption of the control sample and the dilutions a linear regression was performed for the calculation of the cell densities in all other samples measured spectrophotometrically during the test. The inhibition of algal growth was detrmined from: (i) the average specific growth rate, (ii) the yield = the mean cell density at the end of the test minus the mean cell density at the start of the test.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 3.2
- Range finding study
- Test concentrations of the range finding study: limit test at 1000 mg/L
- Results used to determine the conditions for the definitive study: no growth inhibition during the preliminary limit test at 1000 mg/L. - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- ca. 1 080 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 926-1391
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- ca. 1 228.67 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- act. ingr.
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- ca. 387 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 178-522
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- ca. 440.27 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 100 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 113.77 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- act. ingr.
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- ca. 316 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- ca. 359.5 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- act. ingr.
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- ca. 463 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Remarks on result:
- other: 400-543
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- ca. 526.73 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- ca. 199 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Remarks on result:
- other: 141-245
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- ca. 226.39 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 11.38 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- ca. 32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- ca. 36.41 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Details on results:
- Since 90% and 0% of nominal concentration of MGN were measured at 0 and 72h respectively, an additional test monitoring the test substance for 72 hours under light and obscurity conditions, with or without algae, was conducted (see endpoint sumary record in 5.1). The results confirmed that results of the current study could be based on nominal concentrations (please refer to IUCLID section 5.1.3).
- Results with reference substance (positive control):
- - 72h-EC50= 2.305 mg/L (last test performed in 2010)
No further data. - Reported statistics and error estimates:
- Based on the calculated cell densities, the 72-hour ErC50 and the 72-hour EyC50, the corresponding EC10 values and where possible their 95%-confidence limits were calculated by Probit analysis.
For the determination of the 72-hour LOEC and the 72-hour NOEC, the calculated growth rates and yields at each test concentration were tested for significant differences compared to the control values by the Welch t-test (growth rate) and the William t-test (yield), respectively.
Based on the results of this statistical evaluation, the 72-hour NOEC and LOEC for growth rate were not determinable. Therefore, both values were directly determined by visual inspection of the raw data.
Statistics were performed using ToxRat Professional Version 2.10, ToxRat (R) Solutions GmbH. - Validity criteria fulfilled:
- yes
- Conclusions:
- Based on nominal concentrations, the toxicity of the test substance on algae growth rate was calculated as ErC50(72h) = 1080 mg/L, NOErC(72h) = 100 mg/L and the associated LOErC(72h) = 316 mg/L.
- Executive summary:
Toxicity of the substance to Pseudokirchneriella subcapitata has been tested in an algal growth inhibition test, following OECD guideline 201 and EC N°761/2009 C.3 under GLP conditions.
A stock solution of 1000 mg/L was prepared by direct addition into test water. Five different test concentrations (10, 32, 100, 316, and 1000 mg/L) were prepared in triplicates by serial dilution into OECD test water. Six replicates of control solution were prepared only with test water. At the beginning of the test 5 000 cell/mL were inoculated in each test vessels and light intensity was set at an average of 5892 lux. Chemical monitoring were performed through a dedicated UV-HPLC method at 0H and 72h in each test groups.
After 72 hours of exposure, no test substance was measured in the test groups. Thus an additional study was conducted in order to monitor the test substance, through UV-HPLC and TOC analyses, at 0h, 24h, 48h, and 72h under light/no light and algae/no algae conditions. This additional study showed TOC remained constant throughout the experimental phase. However, as measured by UV-HPLC, the test substance decreased showing evidence for a transformation during the test. As results were the same under both light conditions and only occurred when algae were present, external metabolization by algae is assumed without photodegradation.
This observation figured out to base algae results on nominal concentrations (see corresponding endpoint summary record).
Based on nominal concentrations, the toxicity of the test substance on algae growth rate was calculated as ErC50(72h) = 1080 mg/L, NOErC(72h) = 100 mg/L and the associated LOErC(72h) = 316 mg/L.
Table: Results for the determination of the test item in the test samples.
Sample description | test item | calculated | |||||
age | found | test item | nominal | % of | |||
[mg/L] | day | [h] | [mg/L] | D.F. | [mg/L] | [mg/L] | nominal |
control | 0 | 0 | <LOD | 1 | n.a. | 0.000 | n.a. |
control | 3 | 72 | <LOD | 1 | n.a. | 0.000 | n.a. |
10 | 0 | 0 | 9.143 | 1 | 9.143 | 10.000 | 91 |
10 | 0 | 0 | 9.198 | 1 | 9.198 | 10.000 | 92 |
10 | 3 | 72 | <LOD | 1 | n.a. | 10.000 | n.a. |
10 | 3 | 72 | <LOD | 1 | n.a. | 10.000 | n.a. |
32 | 0 | 0 | 31.593 | 1 | 31.593 | 32.000 | 99 |
32 | 0 | 0 | 31.68 | 1 | 31.680 | 32.000 | 99 |
32 | 3 | 72 | <LOD | 1 | n.a. | 32.000 | n.a. |
32 | 3 | 72 | <LOD | 1 | n.a. | 32.000 | n.a. |
100 | 0 | 0 | 10.639 | 10 | 106.390 | 100.000 | 106 |
100 | 0 | 0 | 10.342 | 10 | 103.420 | 100.000 | 103 |
100 | 3 | 72 | <LOD | 10 | n.a. | 100.000 | n.a. |
100 | 3 | 72 | <LOD | 10 | n.a. | 100.000 | n.a. |
320 | 0 | 0 | 30.467 | 10 | 304.670 | 320.000 | 96 |
320 | 0 | 0 | 28.763 | 10 | 287.630 | 320.000 | 91 |
320 | 3 | 72 | <LOD | 10 | n.a. | 320.000 | n.a. |
320 | 3 | 72 | <LOD | 10 | n.a. | 320.000 | n.a. |
1000 | 0 | 0 | 12.102 | 100 | 1210.200 | 1000.000 | 121 |
1000 | 0 | 0 | 9.178 | 100 | 917.800 | 1000.000 | 92 |
1000 | 3 | 72 | <LOD | 100 | n.a. | 1000.000 | n.a. |
1000 | 3 | 72 | <LOD | 100 | n.a. | 1000.000 | n.a. |
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 010
- Report date:
- 2010
Materials and methods
- Study type:
- other: photodegradation during an OECD 201 algal test
Test guideline
- Qualifier:
- no guideline available
- Principles of method if other than guideline:
- Test design:
- 2 concentrations were tested: 100 and 500 mg/L
- 1 replicate with algae in the light (per conc.)
- 1 replicate with algae in the dark (per conc.)
- 1 replicate without algae in the light (per conc.)
- 1 replicate without algae in the dark (per conc.)
- Each replicate was divided in 5 test beakers with 50 mL each
- Light exclusion was achieved by completely coverage of the test beakers in the dark. They were completely wrapped in aluminium foil.
- All beakers were incubated at exactly the same conditions used in a normal Algae test
- Sampling after 0, 24, 48, and 72h
- Per sampling one beaker of each replicate was sacrificed (except at 0h where only one beaker representative for all replicates was sacrificed)
- Standby analysis of the test item after 0, 24, 48, 72 h with HPLC
- calibration with test item
- fortified samples with test item
- Standby analysis of total carbon content after 0, 24, 48, 72 h with TOC
- calibration with test item
- fortified samples with test item - GLP compliance:
- no
Test material
- Reference substance name:
- 2-methylglutaronitrile
- EC Number:
- 224-923-5
- EC Name:
- 2-methylglutaronitrile
- Cas Number:
- 4553-62-2
- Molecular formula:
- C6H8N2
- IUPAC Name:
- 2-methylpentanedinitrile
Constituent 1
Study design
- Radiolabelling:
- no
- Analytical method:
- high-performance liquid chromatography
- other: Total Organic Carbon (TOC) analysis
- Details on sampling:
- - Sampling intervals: Both 2-methylglutaronitrile (MGN) and TOC were analysed after 0, 24, 48 and 72 hours.
- Sampling method: Each replicate of each treatment was divided in 5 test beakers intended to perform the analyses at the different times. Per sampling time, one beaker of each replicate was sacrificed (except at 0h where only one beaker reprsentative for all replicates was sacrificed).
No further data. - Buffers:
- No data
- Light source:
- other: As for the algae study (OECD 201)
- Details on light source:
- Neon ligth tubes as for a typical algae study (OECD 201). For futher details on light conditions, please refer to IUCLID section "6.1.5 Toxicity to aquatic algae and cyanobacteria" and the ESR relative to the study of Hoffmann & Deierling (2010).
In the case of treatment performed in the dark, light exclusion was achieved by completely coverage of the test beakers in the dark. They were completely wrapped in aluminium foil.
- Details on test conditions:
- TEST SYSTEM
- Type, material and volume of test apparatus/vessels: 50 mL test beakers
No further data.
TEST MEDIUM
The same medium as in the algal test was used. For futher details, please refer to IUCLID section "6.1.5 Toxicity to aquatic algae and cyanobacteria" and the ESR relative to the study of Hoffmann & Deierling (2010).
REPLICATION
- No. of replicates: 8 different treatments were applied:
* 100 mg/L MGN - With Algae - In the light: 1 replicate
* 100 mg/L MGN - With Algae - In the dark: 1 replicate
* 100 mg/L MGN - Without Algae - In the light: 1 replicate
* 100 mg/L MGN - Without Algae - In the dark: 1 replicate
* 500 mg/L MGN - With Algae - In the light: 1 replicate
* 500 mg/L MGN - With Algae - In the dark: 1 replicate
* 500 mg/L MGN - Without Algae - In the light: 1 replicate
* 500 mg/L MGN - Without Algae - In the dark: 1 replicate
Duration of test at given test conditionopen allclose all
- Duration:
- 72 h
- Initial conc. measured:
- 100 mg/L
- Duration:
- 72 h
- Initial conc. measured:
- 500 mg/L
- Reference substance:
- no
- Dark controls:
- yes
- Computational methods:
- No data
Results and discussion
- Transformation products:
- not specified
- Details on results:
- TOC analysis:
Regardless the treatment applied (100 or 500 mg/L, light or darkness, algae or not), the TOC remained constant over time.
HPLC analysis of MGN:
Regardless the initial concentration and the light conditions, a decrease in the MGN concentration over time was observed when the test occurred in the presence of algae. The rate of disappearance reached 100% at the end of the 72-hour exposure (except for the treatment with an initial concentration of 500 mg/L and no light). In the absence of algae, such decrease was not observed and the MGN concentrations remained stable over time.
Any other information on results incl. tables
Table: Results for the determination of the test item in the test samples (HPLC)
Sample description | test item | calculated | |||||||
age | found | test item | nominal | % of | |||||
[mg/L] | day | [h] | light | algae | [mg/L] | D.F. | [mg/L] | [mg/L] | nominal |
100 | 0 | 0 | 98.308 | 1 | 98.308 | 100.000 | 98 | ||
100 | 1 | 24 | X | X | 90.789 | 1 | 90.789 | 100.000 | 91 |
100 | 1 | 24 | X | 100.663 | 1 | 100.663 | 100.000 | 101 | |
100 | 1 | 24 | X | 94.206 | 1 | 94.206 | 100.000 | 94 | |
100 | 1 | 24 | 100.874 | 1 | 100.874 | 100.000 | 101 | ||
100 | 2 | 48 | X | X | -1.851 | 1 | -1.851 | 100.000 | -2 |
100 | 2 | 48 | X | -1.831 | 1 | -1.831 | 100.000 | -2 | |
100 | 2 | 48 | X | 101.379 | 1 | 101.379 | 100.000 | 101 | |
100 | 2 | 48 | 101.477 | 1 | 101.477 | 100.000 | 101 | ||
100 | 3 | 72 | X | X | 0.137 | 1 | 0.137 | 100.000 | 0 |
100 | 3 | 72 | X | 0.233 | 1 | 0.233 | 100.000 | 0 | |
100 | 3 | 72 | X | 99.24 | 1 | 99.240 | 100.000 | 99 | |
100 | 3 | 72 | 100.422 | 1 | 100.422 | 100.000 | 100 | ||
500 | 0 | 0 | 100.517 | 5 | 502.585 | 500.000 | 101 | ||
500 | 1 | 24 | X | X | 101.716 | 5 | 508.580 | 500.000 | 102 |
500 | 1 | 24 | X | 101.844 | 5 | 509.220 | 500.000 | 102 | |
500 | 1 | 24 | X | 102.765 | 5 | 513.825 | 500.000 | 103 | |
500 | 1 | 24 | 102.495 | 5 | 512.475 | 500.000 | 102 | ||
500 | 2 | 48 | X | X | 83.827 | 5 | 419.135 | 500.000 | 84 |
500 | 2 | 48 | X | 99.049 | 5 | 495.245 | 500.000 | 99 | |
500 | 2 | 48 | X | 103.703 | 5 | 518.515 | 500.000 | 104 | |
500 | 2 | 48 | 103.34 | 5 | 516.700 | 500.000 | 103 | ||
500 | 3 | 72 | X | X | 0 | 5 | 0.000 | 500.000 | 0 |
500 | 3 | 72 | X | 90.248 | 5 | 451.240 | 500.000 | 90 | |
500 | 3 | 72 | X | 101.399 | 5 | 506.995 | 500.000 | 101 | |
500 | 3 | 72 | 101.735 | 5 | 508.675 | 500.000 | 102 |
Applicant's summary and conclusion
- Validity criteria fulfilled:
- not applicable
- Conclusions:
- The substance appears to be metabolized by algae and no photo-transformation seems to occur. This is the reason why the TOC remains constant.
- Executive summary:
Due to the disappearance of the substance in a previous algae study (Hoffmann & Deierling, 2010), a specific analytical experiment was conducted in order to understand the fate of the test substance in the study and conclude scientifically.
Two different concentrations of the substance were tested (100 and 500 mg/L), with or without algae, in the light or in the dark (1 replicate for each concentration, for each experimental condition); every other experimental conditions remaining the same as for the algae test (Hoffmann & Deierling, 2010). The test substance was monitored by UV-HPLC as detailed in Hoffmann & Deierling (2010) along with total organic carbon (TOC) after 0, 24, 48, and 72 h. Total organic carbon remained constant throughout the test; showing the test substance was still in the test water. However, as measured by UV-HPLC, the test substance decreased showing evidence for a transformation during the test. As results were the same under both light conditions and only occurred when algae were present, external metabolization by algae is assumed without photodegradation.
The results suggest therefore that algae metabolized the test substance whatever the light conditions and that the test item remained in the medium. Thus algae results could be based on nominal concentrations.
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