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EC number: 2252185  CAS number: 4724485
 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
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 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
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 Dissociation constant
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 Additional physicochemical information
 Additional physicochemical properties of nanomaterials
 Nanomaterial agglomeration / aggregation
 Nanomaterial crystalline phase
 Nanomaterial crystallite and grain size
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 Endpoint summary
 Stability
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 Transport and distribution
 Environmental data
 Additional information on environmental fate and behaviour
 Ecotoxicological Summary
 Aquatic toxicity
 Endpoint summary
 Shortterm toxicity to fish
 Longterm toxicity to fish
 Shortterm toxicity to aquatic invertebrates
 Longterm 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
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 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 20112707 to 20130319
 Reliability:
 2 (reliable with restrictions)
 Rationale for reliability incl. deficiencies:
 other: Study conducted in compliance with international standard guidelines under GLP conditions. The pH values of the poststudy are missing in the final report
Data source
Reference
 Reference Type:
 study report
 Title:
 Unnamed
 Year:
 2012
 Report date:
 2013
Materials and methods
Test guidelineopen allclose all
 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 (incl. QA statement)
 Remarks:
 20110831
Test material
 Reference substance name:
 Octylphosphonic acid
 EC Number:
 2252185
 EC Name:
 Octylphosphonic acid
 Cas Number:
 4724485
 Molecular formula:
 C8H19O3P
 IUPAC Name:
 octylphosphonic acid
 Test material form:
 other: granular solid
 Details on test material:
  Name of test material (as cited in study report):Octylphosphinic acid (OPA)
Constituent 1
Sampling and analysis
 Analytical monitoring:
 yes
 Details on sampling:
 Samples were taken from the control (replicate R1R6 pooled) and each test group (replicates R1R3 pooled) at 0 and 72 hours for quantitative analysis. All ohour samples were stored at approximately 20°C prior to analysis. Duplicate samples were taken at 0 and 72 hours and stored at approximately 20°C for further analysis if necessary
Test solutions
 Vehicle:
 no
 Details on test solutions:
 Amount of test item (100 and 32 mg) were each separely dissolved in the culture medium with the aid of ultrasonication for approximately 30 minutes and the volumes adjusted to 1 litter to give a stock solutions of 100 and 32 mg/L. A series of dilutions was made from these stock solutions of 100 mg and 32 mg/L. A serie of dilutions was made from these stock solutions to give further stock solutions of 10, 3.2 and 1.0 mg/L. An aliquot (900 mL) of each of the stock solutions was separately inoculated with algal suspension (11.7 mL) to give the required test concentration of 1.0; 3.2, 10, 32 and 100 mg/L.
The stock solutions and each of the prepared concentrations were inverted several times to ensure adequate mixing and homogeneity.
The concentrations and stability of the test item in the test preparation were verified by chemical analysis at 0 and 72 hours.
Test organisms
 Test organisms (species):
 Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
 Details on test organisms:
 TEST ORGANISM
 Common name:Pseudokirchneriella subcapitata
 Strain:CCAP 278/4
 Source (laboratory, culture collection):Liquids cultures were obtained from the culture collection of Algae and protozoa (CCAP°, SAMS research services Ltd, Scottish Marine Institute, Oban, Argyll Scotland
 Age of inoculum (at test initiation): No data
 Method of cultivation: Masters cultures were maintained in the laboratory by the periodic replenishment of culture medium, and maintained under constant aeration and illumination at 21°C ±1°C
Prior to the start of the test sufficient master culture was added to approximately 100 mL volumes of culture media contained in conical flasks to give an initial cell density of approximately 10e3 cells/mL. The flask were plugged with polyurethane foam stoppers and kept under constant agitation by orbital shaker (100150 rpm) and constant illumination at 24°C ± 1°C until algal cell density was approximately 10e410e5 cells/mL.
Preculture conditions gave an algal suspension in log phase growth characterized by a cell density of 3.83 x 10e5 cells per mL. Inoculation of 900 mL of test medium with 11.7 mL of this algal suspension gave an initial nominal cell density of 5 x 10e3 cells per mL and had no significant dilution effect on the final test concentration.
Study design
 Test type:
 static
 Water media type:
 freshwater
 Limit test:
 no
 Total exposure duration:
 72 h
Test conditions
 Hardness:
 No data
 Test temperature:
 24°C ± 1°C
 pH:
 The pH values of the control culture was observed to increase from pH = 8.2 at ohour to pH = 8.3 at 72hour . The pH deviation in the control cultures was less than 1.5 units after 72 hours and therefore was within the limits given in the guidelines
A concentration dependent decline in pH was observed with increasing test concentration at 0 hours in the range of pH = 8.1 at 1.0 mg/L through to pH = 3.7 at 100 mg/L. This was considered to be due to an intrinsic property of the test item and hence no adjustment was made to the pH prior to exposure  Dissolved oxygen:
 No data
 Salinity:
 not applicable
 Nominal and measured concentrations:
 Nominal concentration: 0, 1.0, 3.2, 10, 32, 100 mg/L
Measured concentration at 0h: 0.745, 0.677, 2.85, 9.15, 30.7, 92.2 mg/L (*)
Measured concentration at 72h: 0, 0.916, 3.05, 9.73, 32.7, 103 mg/L
(*): at 0h, analysis of the sample showed a measured test concentration of 75% of nominal concentration. Analysis of duplicate sample showed a measured concentration of 68% of nominal concentration. Inspection of the data could find no cause for low measured test concentration. Given that a concentration of 92% of nominal concentration was observed at 72 hours, it was considered that the result obtained from ohour analysis were erroneous. This was considered to have had no adverse effect on the outcome of the test given that the 1.0 mg/L test concentration was below the NOEC.
As such,it was considered appropriate to calculate the results based on nominal test concentration only  Details on test conditions:
 TEST SYSTEM
 Test vessel: 250 mL glass conical flask
 Type (delete if not applicable): closed (Polyurethane foam bungs)
 Material, size, headspace, fill volume: fill at 100 mL
 Aeration: constantly shaken at approximately 150 rpm during 72h
 Initial cells density: 5x 10e3
 Control end cells density: At 0h, 24h, 47h, 72h
 No. of vessels per concentration (replicates): 3
 No. of vessels per control (replicates): 6
 Incubation in INFORS Multitron version 2 incubator
GROWTH MEDIUM
Medium used as describe in guideline
TEST MEDIUM / WATER PARAMETERS
 Source/preparation of dilution water: reverse osmosis purified deionized water (Elga Optima 15 + or Elga Purelab Option R15 BP)
OTHER TEST CONDITIONS
 Sterile test conditions: yes/no
 Adjustment of pH: yes. The pH was adjusted at 7.5 ± 0.1 for the culture medium prior to introduce test item
 Photoperiod: constant illumination
 Light intensity and quality: approximately 7000 lux provided by warm white lighting (380730 nm)
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
 Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]
 Chlorophyll measurement:
 Other:
TEST CONCENTRATIONS
 Spacing factor for test concentrations:10
 Justification for using less concentrations than requested by guideline:
 Range finding study: YES
 Test concentrations: 0.1, 1.0, 10 and 100 mg/L
 Results used to determine the conditions for the definitive study: No effects were observed on growth at 0.1, 1 and 10 mg/L. However, growth was observed to be reduced at 100 mg/L  Reference substance (positive control):
 yes
 Remarks:
 potassium dichromate
Results and discussion
Effect concentrationsopen allclose all
 Duration:
 72 h
 Dose descriptor:
 EC10
 Effect conc.:
 23 mg/L
 Nominal / measured:
 nominal
 Conc. based on:
 test mat.
 Basis for effect:
 growth rate
 Duration:
 72 h
 Dose descriptor:
 EC20
 Effect conc.:
 28 mg/L
 Nominal / measured:
 nominal
 Conc. based on:
 test mat.
 Basis for effect:
 growth rate
 Duration:
 72 h
 Dose descriptor:
 EC50
 Effect conc.:
 40 mg/L
 Nominal / measured:
 nominal
 Conc. based on:
 test mat.
 Basis for effect:
 growth rate
 Remarks on result:
 other: 3444 mg/L
 Duration:
 72 h
 Dose descriptor:
 EC10
 Effect conc.:
 12 mg/L
 Nominal / measured:
 nominal
 Conc. based on:
 test mat.
 Basis for effect:
 biomass
 Duration:
 72 h
 Dose descriptor:
 EC20
 Effect conc.:
 16 mg/L
 Nominal / measured:
 nominal
 Conc. based on:
 test mat.
 Basis for effect:
 biomass
 Duration:
 72 h
 Dose descriptor:
 EC50
 Effect conc.:
 23 mg/L
 Nominal / measured:
 nominal
 Conc. based on:
 test mat.
 Basis for effect:
 biomass
 Remarks on result:
 other: 2026 mg/L
 Details on results:
 Cell density values determined at each sapling time and pH values at 0 and 72 hours are given in table 2.
Daily specific growth rate for the control cultures are given in table 3. Growth rate and yield values for the control and test cultures after 72 hours and percentage inhibition values are given in table 4.
Validation criteria:
The following data show that the cell concentration of the control cultures increased by a factor of 176 after 72 hours. This increase was in line with the OECD guideline that states the enhancement must be at least by a factor of 16 after 72 hours
Mean cell density of control at 0 hours : 2.50 x 10e3 cells per mL
Mean cell density of control at 72 hours : 9.28 x 10e5 cells per mL
The mean coefficient of variation for section by section specific growth rate for the control cultures was 12% and hence satisfied the validation criterion given in the OECD Guideline which states the mean must not exceed 35%
The coefficient of variation for average specific growth rate for the control cultures over the test period (072h) was 3% and hence satisfied the validation criterion given in the OECD Guideline which states that this must not exceed 7%.  Results with reference substance (positive control):
 Exposure of Pseudokirchneriella subcapitata (CCAP 278/4) gave the following results:
ErC50 (072h): 1.4 mg/L (1.21.7 mg/L)
EyC50 (072h): 0.59 mg/L (0.530.65 mg/L)
NOEC growth rate: 0.25 mg/L
NOEC yield: 0.25 mg/L
LOEC growth rate: 0.50 mg/L
LOEC yield: 0.50 mg/L
The results were within the normal ranges for this reference item  Reported statistics and error estimates:
 One way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981) and Dunnett's multiple comparison procedure for comparing severals treatments with a control (Dunnett, 1955) was carried out on the growth rate and yield data after 72 hours for the control and all test concentration to determine any statistical significant differences between the test and the control groups. All statistical analyses were performed using the SAS computer software package (SAS 19992001)
Any other information on results incl. tables
Table 2: Cell Densities and pH values in the definitive test
Nominal concentration (mg/L) 
pH 
Cell densities * (cells per mL) 
pH 

0h 
0h 
24h 
47h 
72h 
72h 

Control 
R1 
8.2 
5.04.E+3 
3.02.E+4 
1.49.E+5 
7.24.E+5 
8.3 
R2 
5.19.E+3 
3.00.E+4 
1.74.E+5 
9.36.E+5 

R3 
5.02.E+3 
2.73.E+4 
1.71.E+5 
8.55.E+5 

R4 
5.82.E+3 
3.46.E+4 
1.75.E+5 
1.06.E+6 

R5 
5.09.E+3 
3.12.E+4 
1.34.E+5 
8.83.E+5 

R6 
5.04.E+3 
3.51.E+4 
1.21.E+5 
1.11 E+6 

Mean 
5.20.E+3 
3.14.E+4 
1.54.E+5 
9.28.E+5 

1.0 
R1 
8.1 
5.23.E+3 
3.16.E+4 
1.64.E+5 
1.00 E+6 
8.2 
R2 
5.06.E+3 
3.38.E+4 
1.95.E+5 
1.03 E+6 

R3 
5.38.E+3 
3.18.E+4 
1.50.E+5 
1.01 E+6 

Mean 
5.23.E+3 
3.24.E+4 
1.69.E+5 
1.01 E+6 

3.2 
R1 
7.9 
5.22.E+3 
3.74.E+4 
1.67.E+5 
8.66.E+5 
8.1 
R2 
5.02.E+3 
3.57.E+4 
1.78.E+5 
1.17 E+6 

R3 
5.09.E+3 
3.02.E+4 
1.43.E+5 
9.19.E+5 

Mean 
5.11.E+3 
3.44.E+4 
1.63.E+5 
9.85.E+5 

10 
R1 
7.7 
5.15.E+3 
3.62.E+4 
1.54.E+5 
9.24.E+5 
7.9 
R2 
5.22.E+3 
3.69.E+4 
1.74.E+5 
8.30.E+5 

R3 
5.06.E+3 
3.83.E+4 
1.90.E+5 
8.92.E+5 

Mean 
5.14.E+3 
3.72.E+4 
1.72.E+5 
8.82.E+5 

32 
R1 
7.3 
5.30.E+3 
2.26.E+4 
7.85.E+4 
3.32.E+5 
7.3 
R2 
5.14.E+3 
1.98.E+4 
5.07.E+4 
1.79.E+5 

R3 
5.30.E+3 
1.93.E+4 
4.45.E+4 
1.21.E+5 

Mean 
5.25.E+3 
2.05.E+4 
5.79.E+4 
2.11.E+5 

100 
R1 
3.7 
5.06.E+3 
1.40.E+4 
5.38.E+3 
6.98.E+3 
3.5 
R2 
5.03.E+3 
1.55.E+4 
1.12.E+4 
4.01.E+3 

R3 
5.07.E+3 
1.66.E+4 
7.54.E+3 
6.41.E+3 

Mean 
5.06.E+3 
1.54.E+4 
8.04.E+3 
5.80.E+3 
* cell densities represent the mean number of cells per mL calculated from the mean of the cell counts from 3 counts for each of the replicate flasks.
R1R6 = Replicate 1 to 6
Table 3: Daily specific growth rates for the control cultures in the definitive test

Daily specific growth rate (cells/mL/hour) 

Day 01 
Day 12 
Day 23 

Control 
R1 
0.075 
0.069 
0.063 
R2 
0.075 
0.076 
0.067 

R3 
0.071 
0.080 
0.064 

R4 
0.081 
0.071 
0.072 

R5 
0.076 
0.063 
0.075 

R6 
0.081 
0.054 
0.089 

Mean 
0.077 
0.069 
0.072 
R1R6 = Replicate 1 to 6
Table 4: Inhibition of growth rate and yield in the definitive test
Nominal concentration (mg/L) 
Growth rate (cell/mL/hour) 
Yield (Cell/mL) 

072 h 
% Inhibition + 
072 h 
% Inhibition +* 

Control 
R1 
0.069 

7.19.E+05 

R2 
0.073 

9.31.E+05 


R3 
0.071 

8.50.E+05 


R4 
0.074 
 
1.06.E+06 
 

R5 
0.074 

8.78.E+05 


R6 
0.072 

1.11.E+06 


Mean 
0.075 

9.23.E+05 


SD 
0.002 

1.42.E+05 


1.0 
R1 
0.074 
[3] 
9.99.E+05 

R2 
0.074 
[3] 
1.03.E+06 


R3 
0.074 
[3] 
1.00.E+06 


Mean 
0.074 
[3] 
1.01.E+06 
[9] 

SD 
0.000 

1.49.E+04 


3.2 
R1 
0.072 
0 
8.61.E+05 

R2 
0.076 
[6] 
1.16.E+06 


R3 
0.072 
0 
9.14.E+05 


Mean 
0.073 
[2] 
9.80 .E+05 
[6] 

SD 
0.002 

1.62 .E+05 


10 
R1 
0.072 
0 
9.19 .E+05 

R2 
0.071 
1 
8.25 .E+05 


R3 
0.072 
0 
8.87 .E+05 


Mean 
0.072 
0 
8.77 .E+05 
5 

SD 
0.001 

4.77 .E+04 


32 
R1 
0.058 
19 
3.27 .E+05 

R2 
0.050 
31 
1.73 .E+05 


R3 
0.044 
39 
1.16 .E+05 


Mean 
0.051 
30 
2.05 .E+05 
78 

SD 
0.007 

1.09 .E+05 


100 
R1 
0.005 
93 
1.92 .E+03 

R2 
0.003 
104 
1.02 .E+03 


R3 
0.003 
96 
1.34 .E+03 


Mean 
0.002 
98 
7.44 .E+02 
100 

SD 
0.004 

1.56 .E+03 

+ In accordance with the OECD test guideline inhibition of growth was calculated based on the 0 Hour measured cell densities whilst inhibition of yield was calculated based on a nominal cell density of 5.00 E+03 cells/mL
* In accordnace with the OECD test guideline only the mean value for yield for each test concentration is calculated
R1  R6 = Replicates 1 to 6
[Increase in growth as compared to controls]
Table 5: Cell Densities and percentage inhibition of Growth from the poststudy experiment
Nominal Concentration (mg/L) 
Cell Densities * (Cells per mL) 
Inhibition Values (%)** 

0 hours 
72 hours 
Growth Rate 
Yield 

Control 
R1 
5.68 E+03 
6.68 E+05 
 
 
R2 
5.01 E+03 
4.67 E+05 

Mean 
5.34 E+03 
5.67 E+05 

1.0 
R1 
5.23 E+03 
7.95 E+05 
[6] 
[31] 
R2 
5.17 E+03 
6.87 E+05 

Mean 
5.20 E+03 
7.41 E+05 

10 
R1 
5.38 E+03 
5.70 E+05 
[2] 
[12] 
R2 
5.26 E+03 
6.95 E+05 

Mean 
5.32 E+03 
6.32 E+05 

100 
R1 
7.14 E+03 
5.89 E+04 
57 
92 
R2 
6.66 E+03 
4.30 E+04 

Mean 
6.90 E+03 
5.10 E+04 
*Cell densities represent the mean number of cell per mL calculated from the mean of the cell counts from 3 counts for each of the replicate flasks.
**Inhibition values for growth rate and yield determined based on the 0 hour measured cell densities
R1 and R2 = Replicate 1 and Replicate 2
[Increase in growth compared to controls]
Applicant's summary and conclusion
 Validity criteria fulfilled:
 yes
 Conclusions:
 The effect of the test item on the growth of Pseudokirchneriella subcapitata has been investigated over a 72hour period and gave the following result
s:
ErC10 : 23 mg/L
ErC50 : 40 mg/L with 95% confidence limits of 3544 mg/L
EyC50 : 23 mg/L with 95% confidence limits of 2026 mg/L
NOEC (growth rate) = 10 mg/L
NOEC (Yield) = 10 mg/L
LOEC Growth rate: 32 mg/L
LOEC Yield = 32 mg/L  Executive summary:
In a 72 hour acute toxicity study, the cultures of Pseudokirchneriella subcapitata were exposed to OPA at nominal concentrations of 1.0, 3.2, 10, 32 and 100 mg/L under static conditions in accordance with the OECD 201, after a range finding study at nominal concentration of 0.1, 1.0, 10 and 100 mg/L
Analytical verification showed an acceptable correlation between the nominal and the measured concentration.
The results were based on nominal concentration
The following results were determined:
NOEC Growth Rate and yield = 10 mg/L
EC10 Growth rate= 23 mg/L
EC 50 Growth rate= 40 mg/L with confidence limits of 35 44 mg/L
EC 50 Yield = 23 mg/L with confidence limits of 20 26 mg/L
All validity criteria were met, the test can be considered to be valid
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