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EC number: 202-860-4 | CAS number: 100-52-7
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
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- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
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- Auto flammability
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- Stability: thermal, sunlight, metals
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- Dissociation constant
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- Endpoint summary
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- 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
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- Genetic toxicity
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- Exposure related observations in humans
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- Additional toxicological data

Toxicity to aquatic algae and cyanobacteria
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2018-12-03 to 2018-12-10
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Version / remarks:
- OECD Guideline No. 201 for Testing of Chemicals (2011)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- Council Regulation (EC) No. 266/2016/Method C.3
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- Test item: Benzaldehyde
CAS No./EC No.: 100-52-7/202-860-4
Batch number: #1803-1
Purity (certified): > 99.5 % wt
Appearance: Clear, colorless to light yellow, liquid
Water solubility: 6.95 g/L at 25 °C
Relative density: 1.041 – 1.046 at 25 °C
Vapor pressure: 169 Pa at 25 °C
Expiry date: 2020-01-15 - Analytical monitoring:
- yes
- Details on sampling:
- Chemical Analysis of Test Item Concentrations
Determination of the test item: The samples were analysed under GLP with a GC-MS method, which has been implemented under non-GLP and documented finally in the GLP raw data.
Sampling schedule: All concentration levels and the control were analytically verified via GC-MS after 0 hours (start of exposure) and 72 hours (end of the exposure) with algae.
Sampling and pre-treatment: Separate replicates for the test item analysis after 0 hours and 72 hours were prepared in the original test vessels with algae at the start of the exposure and incubated under test conditions until analysis. The samples were stabilized immediately after sampling by extraction. The samples were preferably measured within one day after sampling.
Biological Parameters
Chlorophyll a-fluorescence: The cell density was measured daily via Chlorophyll a-fluorescence, excitation at 436 nm, emission at 685 nm. Dilution water was used as a background signal. No self-fluorescence was observed in the preliminary range finding test at the 100 mg/L test item solution.
Microscopic evaluation: Microscopic evaluation of the cells at the start and the end of the incubation period was carried out. The cells were checked for unusual cell shapes, colour differences, differences in chloroplast morphology, flocculation, adherence of algae to test containers and agglutination of algae cells.
Physical Chemical Properties
The pH-values at the start of exposure were measured in one additional replicate of each concentration level and the control. At the end of exposure, the pH-values were measured from pooled samples of the test item concentrations and the control. The room temperature was measured continuously. Light intensity was measured prior to the start of exposure.
Equipment
pH-Meter, pH 191 (WTW)
Hygro-thermograph (KLIMATHERM)
Digital Lux meter (TESTO)
Fluorometer: Microplate Reader Chameleon V (HIDEX) with Software Micro Win 4.41 (MIKROTEC LABORSYSTEME GMBH) for determination of the cell density
Shaker, SM 25 A (BÜHLER)
Light microscope (ZEISS Lab 16)
Fluorescent tubes, OSRAM L 36 W/865, cool daylight (test), OSRAM L 30 W/840, cool white energy saver (culture)
Piston Stroke Pipettes (THERMO SCIENTIFIC, BRANDT)
Analytical balance, LA230-S (SARTORIUS)
Balance, PLS 1200-3A (KERN) - Vehicle:
- no
- Details on test solutions:
- Test item: Benzaldehyde
Stock solution, media preparation: A stock solution with a nominal test item concentration of 50.0 mg/L was freshly prepared with dilution water prior to the start of exposure. The stock solution was agitated until the solution was visually clear.
Test concentrations (nominal): Seven concentration levels were tested in a geometrical series with (nominal) dilution factor of √10: 0.0500 - 0.158 - 0.500 - 1.58 - 5.00 - 15.8 - 50.0 mg/L, corresponding to the geometric mean measured test item concentrations: 0.0140 – 0.0235 – 0.0867 – 0.104 – 0.174 – 7.28 – 42.4 mg/L. The concentration levels are based on the results of a preliminary range finding test.
Control: Six replicates (without test item) were tested under the same test conditions as the test item replicates.
Reference Item: Potassium dichromate was tested as a reference item.
Replicates: Three replicates per concentration level and six for the control were tested. Separate replicates for each measuring time were prepared.
Test containers: Sterile headspace flasks, volume: 59 mL, with aluminium tops with PTFE seals
Test volume: 59 mL - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- Test organism: Pseudokirchneriella subcapitata HINDÁK CCAP 278/4
Synonyms: Selenastrum capricornutum; Ankistrodesmus subcapitata; Raphidocelis subcapitata; Ankistrodesmus bibraianus (Experimental Phycology and Culture Collection of Algae at the University of Goettingen 2014)
Reason for the selection of the test organism: Pseudokirchneriella subcapitata is a suitable green alga species according to the guideline.
Origin: Culture Collection of Algae and Protozoa (CCAP)
SAMS Research Services Ltd
Dunstaffnage Marine Laboratory
Dunbeg, OBAN; Argyll PA37 1QA; Scotland, UK
Cultivation at test facility: Fresh stocks are prepared every month on Z-Agar. Light intensity amounts 2567 – 5130 lux for 24 hours per day.
Culture medium: Nutrient medium Z according to LÜTTGE et al. (1994).
Preculture: A four days old preculture, prepared in dilution water and incubated under test conditions, was used as inoculum.
Initial cell density Nominal: approximately 5 x 103 - 104 cells/mL
Actual: 6442 cells/mL - Test type:
- static
- Test temperature:
- 21 – 24 °C
- Details on test conditions:
- Test method: Static procedure
Performance of the test: A static test was carried out. With regard to the volatility of the test item sealed glass flasks without headspace were used to reduce loss of test item.
Duration of the test: 72 hours
Replicates: Three replicates per concentration level and six for the control were tested. Separate replicates for each measuring time were prepared.
Test containers: Sterile headspace flasks, volume: 59 mL, with aluminium tops with PTFE seals
Test volume: 59 mL
Preculture: A four days old preculture, prepared in dilution water and incubated under test conditions, was used as inoculum.
Initial cell density: Nominal: approximately 5 x 103 - 104 cells/mL
Actual: 6442 cells/mL
Application: Application was carried out by adding appropriate volumes of algae inoculum to the test solutions.
Incubation: The flasks were positioned randomly and repositioned daily.
Temperature (target): Nominal range: 21 – 24 °C, controlled at ± 2 °C
Agitation: Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm.
Light intensity (target): Approximately 4440 to 8880 lux, corresponding to 60 to 120 µE*m-2*s-1
Light regime: 24 hours/day light
Light homogeneity (target): Within ± 15% over incubation area - Reference substance (positive control):
- yes
- Remarks:
- Potassisum dichromate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Remarks:
- growth rate
- Effect conc.:
- 33.1 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Remarks:
- growth rate
- Effect conc.:
- 0.039 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Remarks:
- Yield
- Effect conc.:
- 8.05 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Remarks:
- Yield
- Effect conc.:
- 0.021 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Details on results:
- The toxicity of Benzaldehyde (Batch no.: #1803-1) to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3 from 2018-12-03 to 2018-12-10 at the test facility. The aim of the study was the determination of the effects on growth rate and yield over a period of 72 hours.
The study was conducted under static conditions with an initial cell density of 6442 cells/mL. A stock solution with a nominal concentration of 50.0 mg/L was prepared once prior to the start of the exposure. An appropriate amount of the test item was weighed out and transferred with an appropriate amount of the dilution water into a glass flask. This dispersion was agitated until it was visually clear. Seven concentration levels were tested in a geometrical series with a spacing factor of √10: 0.0500 - 0.158 - 0.500 - 1.58 - 5.00 - 15.8 - 50.0 mg/L, corresponding to the geometric mean measured test item concentrations: 0.0140 – 0.0235 – 0.0867 – 0.104 – 0.174 – 7.28 – 42.4 mg/L. Three replicates were tested for each test item concentration and six replicates for the control. The environmental conditions were within the acceptable limits.
At the start of the exposure the measured concentrations were in the range of 84 to 98% of the nominal values. At the end of exposure after 72 hours, the measured concentrations were in the range of < LOQ to 80% of the nominal test item concentrations. - Results with reference substance (positive control):
- The toxicity of potassium dichromate (SIGMA-ALDRICH, batch number MKBV0900V, purity 99.0 %, CAS RN 7778-50-9) to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined over a period of 72 hours from 2018-09-25 to 2018-09-28 (with headspace) and 2018-07-10 to 2018-07-13 (without headspace), respectively. The reference item toxicity is in the valid range following test facility SOPS.
- Validity criteria fulfilled:
- yes
- Conclusions:
- The 72-hour algal EC50 (growth rate) was 33.1 mg/l and the EC50 (yield) was 8.05 mg/l.
- Executive summary:
The acute toxicity of benzaldehyde to Pseudokirchneriella subcapitata was studied in a 72-hour toxicity study according to OECD 201. The 72-hour algal EC50 (growth rate) was 33.1 mg/l and the EC50 (yield) was 8.05 mg/l.
Reference
Cell Densities
Geometric mean measured test item concentration |
Replicate |
Cell density [cells/mL] |
|||
[mg/L] |
No. |
0 hours |
24 hours |
48 hours |
72 hours |
42.4 |
1 |
6442 |
12487 |
18805 |
33087 |
2 |
6442 |
11385 |
18524 |
38019 |
|
3 |
6442 |
11596 |
19726 |
33561 |
|
Mean |
6442 |
11823 |
19018 |
34889 |
|
7.28 |
1 |
6442 |
14438 |
60431 |
227201 |
2 |
6442 |
17819 |
55625 |
207966 |
|
3 |
6442 |
17636 |
51489 |
261122 |
|
Mean |
6442 |
16631 |
55848 |
232096 |
|
0.174 |
1 |
6442 |
18626 |
55039 |
193548 |
2 |
6442 |
16710 |
55481 |
209972 |
|
3 |
6442 |
14813 |
57532 |
182367 |
|
Mean |
6442 |
16716 |
56017 |
195296 |
|
0.104 |
1 |
6442 |
17201 |
54404 |
229326 |
2 |
6442 |
18346 |
58704 |
167571 |
|
3 |
6442 |
17479 |
50114 |
224470 |
|
Mean |
6442 |
17675 |
54407 |
207122 |
|
0.0867 |
1 |
6442 |
23350 |
93598 |
330791 |
2 |
6442 |
22117 |
82178 |
354300 |
|
3 |
6442 |
21474 |
107321 |
339479 |
|
Mean |
6442 |
22314 |
94366 |
341523 |
|
0.0235 |
1 |
6442 |
29000 |
119738 |
376940 |
2 |
6442 |
26846 |
122410 |
363311 |
|
3 |
6442 |
28010 |
127662 |
437786 |
|
Mean |
6442 |
27952 |
123270 |
392679 |
|
0.0140 |
1 |
6442 |
29867 |
117713 |
403553 |
2 |
6442 |
25108 |
114570 |
393325 |
|
3 |
6442 |
25145 |
117226 |
412291 |
|
Mean |
6442 |
26707 |
116503 |
403056 |
|
Control |
1 |
6442 |
30371 |
124892 |
454916 |
2 |
6442 |
29418 |
136033 |
403900 |
|
3 |
6442 |
29341 |
133611 |
402584 |
|
4 |
6442 |
29517 |
134733 |
410578 |
|
5 |
6442 |
28137 |
127280 |
441907 |
|
6 |
6442 |
30746 |
125627 |
398513 |
|
Mean |
6442 |
29588 |
130363 |
418733 |
Evaluation after 72 hours
Statistically significant differences of growth rates and yield compared to control values are marked (+), not significant differences are marked (-).
Geometric mean measured test item concentration |
Replicate |
Growth Rate |
Inhibition of Growth Rate |
Yield |
Inhibition of Yield |
||
[mg/L] |
No. |
[d-1] |
[%] |
[cells/mL] |
[%] |
||
42.4 |
1 |
|
0.55 |
61 |
|
26645 |
94 |
2 |
|
0.59 |
57 |
|
31577 |
92 |
|
3 |
|
0.55 |
60 |
|
27119 |
93 |
|
Mean |
(+) |
0.56 |
60 |
(+) |
28447 |
93 |
|
7.28 |
1 |
|
1.19 |
15 |
|
220759 |
46 |
2 |
|
1.16 |
17 |
|
201524 |
51 |
|
3 |
|
1.23 |
11 |
|
254680 |
38 |
|
Mean |
(+) |
1.19 |
14 |
(+) |
225654 |
45 |
|
0.174 |
1 |
|
1.13 |
18 |
|
187106 |
55 |
2 |
|
1.16 |
17 |
|
203530 |
51 |
|
3 |
|
1.11 |
20 |
|
175925 |
57 |
|
Mean |
(+) |
1.14 |
18 |
(+) |
188854 |
54 |
|
0.104 |
1 |
|
1.19 |
14 |
|
222884 |
46 |
2 |
|
1.09 |
22 |
|
161129 |
61 |
|
3 |
|
1.18 |
15 |
|
218028 |
47 |
|
Mean |
(+) |
1.15 |
17 |
(+) |
200680 |
51 |
|
0.0867 |
1 |
|
1.31 |
6 |
|
324349 |
21 |
2 |
|
1.34 |
4 |
|
347858 |
16 |
|
3 |
|
1.32 |
5 |
|
333037 |
19 |
|
Mean |
(+) |
1.32 |
5 |
(+) |
335081 |
19 |
|
0.0235 |
1 |
|
1.36 |
2 |
|
370498 |
10 |
2 |
|
1.34 |
3 |
|
356869 |
13 |
|
3 |
|
1.41 |
-1 |
|
431344 |
-5 |
|
Mean |
(-) |
1.37 |
2 |
(-) |
386237 |
6 |
|
0.0140 |
1 |
|
1.38 |
1 |
|
397111 |
4 |
2 |
|
1.37 |
1 |
|
386883 |
6 |
|
3 |
|
1.39 |
0 |
|
405849 |
2 |
|
Mean |
(-) |
1.38 |
1 |
(-) |
396614 |
4 |
|
Control |
1 |
|
1.42 |
|
|
448474 |
|
2 |
|
1.38 |
|
|
397458 |
||
3 |
|
1.38 |
|
|
396142 |
||
4 |
|
1.38 |
|
|
404136 |
||
5 |
|
1.41 |
|
|
435465 |
||
6 |
|
1.37 |
|
|
392071 |
||
Mean |
|
1.39 |
|
|
412291 |
Section-by-Section and Average Specific Growth Rates of the Control Group (0 - 72 hours)
|
Replicate No. |
Specific growth rate [d-1] |
Mean (0 - 72 hours) |
SD ± |
CV |
Mean CV [%] |
||
section-by-section |
||||||||
0 - 24 hours |
24 - 48 hours |
48 - 72 hours |
||||||
Control |
1 |
1.55 |
1.41 |
1.29 |
1.42 |
0.129 |
9.10 |
14.5 |
2 |
1.52 |
1.53 |
1.09 |
1.38 |
0.252 |
18.3 |
||
3 |
1.52 |
1.52 |
1.10 |
1.38 |
0.239 |
17.3 |
||
4 |
1.52 |
1.52 |
1.11 |
1.39 |
0.234 |
16.9 |
||
5 |
1.47 |
1.51 |
1.25 |
1.41 |
0.144 |
10.2 |
||
6 |
1.56 |
1.41 |
1.15 |
1.38 |
0.206 |
15.0 |
||
|
|
|
Mean |
1.39 |
|
|
||
|
|
|
SD ± |
0.02 |
|
|||
|
|
|
CV [%] |
1.33 |
|
SD = Standard deviation CV = Coefficient of variation
Physicochemical Data
The environmental conditions (pH-value, room temperature, light intensity) were determined to be within the acceptable limits in the definitive study. The test media were clear throughout the exposure phase.
Environmental Conditions
Geometric mean measured test item concentration |
pH-values |
||
[mg/L] |
Start; 0 hours |
End; 72 hours |
|
42.4 |
8.24 |
8.31 |
|
7.28 |
8.25 |
8.05 |
|
0.174 |
8.24 |
8.56 |
|
0.104 |
8.21 |
8.80 |
|
0.0867 |
8.18 |
9.16 |
|
0.0235 |
8.15 |
9.16 |
|
0.0140 |
8.16 |
9.14 |
|
Control |
8.19 |
9.10 |
|
Room temperature [°C]: |
min.: 22.5 |
max.: 23.5 |
mean value: 23.0 |
Light intensity |
n = 23 |
mean value: 5744 range of the measured values: 5025 to 6261 equalling -12.5 to 9.00% |
CV [%]: 6.84% |
CV = Coefficient of variation n = number of measuring points
Measured Exposure Concentrations during the Definitive Test
The concentrations of Benzaldehyde and the control were determined in fresh media (0 hours) and at the end of the exposure interval (72 hours) via GC-MS.
At the start of the exposure the measured concentrations were in the range of 84 to 98% of the nominal values. At the end of exposure after 72 hours, the measured concentrations were in the range of < LOQ to 80% of the nominal test item concentrations. The geometric mean measured concentrations are 42.4 – 7.28 – 0.174 – 0.104 – 0.0867 – 0.0235 – 0.0140 mg/L.
Measured Concentration of Benzaldehyde in the Definitive Test with Algae
Sampling date |
Fresh Media, |
Old Media, |
|
||
Nominal test item concentration |
Meas. conc. [mg/L] |
% |
Meas. conc. [mg/L] |
% |
Geometric mean measured concentration1) [mg/L] |
|
Benzaldehyde |
||||
50.0 |
44.8 |
90 |
40.1 |
80 |
42.4 |
15.8 |
14.8 |
94 |
3.58 |
23 |
7.28 |
5.00 |
4.75 |
95 |
0.00640* |
0 |
0.174 |
1.58 |
1.41 |
89 |
0.00774* |
0 |
0.104 |
0.50 |
0.419 |
84 |
0.0179 |
4 |
0.0867 |
0.158 |
0.138 |
87 |
< LOQ |
- |
0.0235 |
0.050 |
0.0490 |
98 |
< LOQ |
- |
0.0140 |
Control |
< LOQ |
- |
< LOQ |
- |
< LOQ |
Meas. conc.= measured concentration of the test item, dilution factors taken into account
% = percent of the nominal measured concentration of the test item
LOQ = limit of quantification of the analytical method (0.008 mg/L of the test item)
1) = Values < LOQ taken into account with ½ LOQ (0.004 mg/L of the test item)
* = < LOQ but > 70% LOQ
NOEC, LOEC, ECx-values ofBenzaldehyde(0 - 72 hours)
(based on the geometric mean measured test item concentrations, with 95% confidence intervals)
|
Growth Rate Inhibition |
NOEC |
0.0235 |
LOEC |
0.0867 |
ErC10 |
0.0389 (0.0258 – 7.62) |
ErC20 |
9.70 (5.79 – 13.3) |
ErC50 |
33.1 (28.2 – 39.5) |
|
Yield Inhibition [mg/L] |
NOEC |
0.0235 |
LOEC |
0.0867 |
EyC10 |
0.0205 (0.0141 – 0.0275) |
EyC20 |
0.0293 (0.0208 – 0.0419) |
EyC50 |
8.05 (0.120 – 16.4) |
EC50-Values of the Reference Item based on nominal concentrations[mg/L], (0-72 hours)
|
with Headspace |
without Headspace |
Valid Range (average ± 3 x SD) |
|
|
Growth Rate Inhibition |
|||
ErC50 |
0.799 |
0.244 |
0.744±0.626 |
|
95 % confidence interval |
0.707 - 1.30 |
0.175 - 0.350* |
||
|
Yield Inhibition |
|
||
EyC50 |
0.607 |
0.236 |
0.409±0.208 |
|
95 % confidence interval |
0.404 - 0.694 |
0.175 - 0.350* |
|
SD = Standard deviation
* = The highest concentration level without statistical significance and the lowest concentration level with statisical significance were used as confidence limits.
Description of key information
The toxicity of benzaldehyde to Pseudokirchneriella subcapitata was studied in a 72-hour toxicity study according to OECD 201. The 72-hour algal EC50 (growth rate) was 33.1 mg/l and the EC50 (yield) was 8.05 mg/l (mean measured concentration). An EC10 (growth) of 0.0389 mg/l and EC10 (yield) of 0.0205 mg/l were also reported.
Supporting studies showed 8-day NOEC (TGK) values of 20 mg/L (Bringmann and Kuhn, 1976) and 34 mg/l (Bringmann and Kuhn 1977; Bringmann and Kuhn, 1980). The outcome of a QSAR (ECOSAR) predicted an EC50 of 23 mg/L.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 8.05 mg/L
- EC10 or NOEC for freshwater algae:
- 0.021 mg/L
Additional information
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