Reaction product of resin acid and rosin acid, 12-Hydroxyoctadecanoic and stearic acid with dipentaerythritol

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 Sept. 2007 - 11 Jan. 2008

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)

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 7-2-A
- Expiration date of the lot/batch: April 2010
- Purity test date: not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature at about 20 °C, away from direct sunlight
- Stability under test conditions: not stated
- Solubility and stability of the test substance in the solvent/vehicle: not stated
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item is a complex mixture containing different sparingly soluble compounds. In order to assess its toxicity, water accomodated fractions (WAF's) were prepared.
- Preliminary purification step (if any): none
- Final dilution of a dissolved solid, stock liquid or gel:
The following WAFs with the loading rates of 0.32, 1.0, 3.2, 10, 32 and 100 mg/l were prepared.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: from each test concentration
- Sampling method: duplicate samples from each test medium (without algae)
- Sample storage conditions before analysis: deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Strain: No. 61.81
- Source (laboratory, culture collection): Collection of Algal Cultures (SAG, Institute for Plant Physiology, University of Göttingen, Göttingen, Germany)
- Age of inoculum (at test initiation): not stated
- Method of cultivation: standardized conditions according to the test guidelines

ACCLIMATION
no acclimation

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Hardness:

0.24

Test temperature:

23-24 °C

pH:

8.2 at start, 9.1 to 9.6 at end

Dissolved oxygen:

not stated

Salinity:

not stated

Conductivity:

not stated

Nominal and measured concentrations:

WAF with nominal loading rate: 0.32, 1.0, 3.2, 10, 32, 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Erlenmeyer flask
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 50 ml flasks, filled with 15 ml algal suspension
- Aeration: none
- Type of flow-through (e.g. peristaltic or proportional diluter): none
- Renewal rate of test solution (frequency/flow rate): none
- Initial cells density: 10,000 algal cells per ml
- Control end cells density: 2079000 algal cells (Mean value)
- No. of organisms per vessel: n/a
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per vehicle control (replicates): n/a

GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used:

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: sterile, purified water
- Total organic carbon: not stated
- Particulate matter: not stated
- Metals: not stated
- Pesticides: not stated
- Chlorine: not stated
- Alkalinity: not stated
- Ca/mg ratio: not stated
- Conductivity: not stated
- Culture medium different from test medium: no
- Intervals of water quality measurement: not stated

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: 24 hours/day
- Light intensity and quality: 8300 Lux
- Salinity (for marine algae): n/a

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: 3.2
- Justification for using less concentrations than requested by guideline:
- Range finding study: none

Reference substance (positive control):

no

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Loading rate

Remarks:

Water accomodated fraction

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Loading rate

Remarks:

Water accomodated fraction

Basis for effect:

biomass

Details on results:

- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): no abnormalities observed (microscopically)
- Unusual cell shape: none
- Colour differences: none
- Flocculation: none
- Adherence to test vessels: none
- Aggregation of algal cells: none

Calculation of coefficient of variations for validity criteria:

biomass at time				Growth rate
0 h	24 h	48 h	72 h	0-24 h	24-48 h	48-72 h
1,40	9,60	66,3	302,4	1,92529086	1,9324268	1,51756075
1,4	8,8	63,9	286,8	1,83827948	1,98256764	1,50146575
1,4	9,7	69,8	284,8	1,93565365	1,97350812	1,40615317
1,4	9,2	68	297,2	1,88273125	2,00030422	1,47489761
1,4	9,1	62,6	295,8	1,87180218	1,92849086	1,55291827
1,4	9,7	69,1	279,3	1,93565365	1,96342885	1,39673174
			Mean	1,89823518	1,96345442	1,47495455
			Standard deviation	0,0401648	0,02831771	0,06236812
			Coefficient of variation of average specific growth rate out of the daily/section by section growth rates
				2,11590204	1,44223935	4,22847769
			mean coefficient of variation of the daily/section by section specific growth rate
				2,59553969

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, Salacos 168 ARV had no inhibitory effect on the growth of Pseudokirchneriella subcapitata up to its solubility limit in test water under the present test conditions.

Executive summary:

The analytically measured test item concentration in the test medium was below limit of quantification (0.3 mg/L) at the beginning and the end of the test period of 72 hours. Since WAFs of the test item with sparingly soluble components were tested, all reported biological results are based on the loading rates.

The test item had no statistically significant inhibitory effect on the growth (yield and growth rate) of Pseudokirchneriella subcapitata during the test period of 72 hours up to and including the highest nominal test concentration of 100 mg/L (results of Dunnett's tests, one sided, Alpha = 0.05).

The test concentration with the loading rate 100 mg/l was, therefore, determined to be the 72 -hour NOEC. This value might even be higher, but concentrations of the test item above the loading rate of 100 mg/l were not tested in accordance with the EU Commission Directive 92/69/EEC. The 72 Hour LOEC and the 72 hour EC10 and EC50 for the algal growth rate (µ) and yield (y) could not be quanttified due to the absence of a toxic effect of Salacos 168 ARV at the tested concentrations. Accordingly, these parameters were clearly higher than the loading rate of 100 mg/L. The microscopic examination of the algal cells after 72 hours exposure showed no difference getween the algae growing at the test concentration of nominal 100 mg/l and the algal cells in the control. The shape and size of the algal cells growing in test media containing the test item at and up to this concentration were obviously not affected.

In the control the biomass increased by a factor of 208 over 72 hours. The validity criterion of increase of biomass by at least a factor of 16 within three days was fulfilled.The mean coefficient of variation of the daily growth rates was 4.2 % during 72 hours. According to the OECD test guideline, the mean coefficient of variation in the control must not be higher than 35 %. Thus, the validity criterion was fulfilled.

The coefficeint of variation of the average specific growth rates in the replicates of the control after 72 hours was 2.6 %. According to the OECD test guideline, the coefficient of variation must not be higher than 7 %. Thus, the validity criterion was fulfilled.

No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions ghroughout the test period.

At the start of the test, the pH values in the test media and the control were 8.2. At the end of the test, pH values of 9.1 to 9.6 were measured. The water temperature during the test was 23 -24 °C.

Description of key information

One Guideline Study performed. Water accomodated Fractions with loading rates up to 100 mg/l tested.

No toxic effects observed up to the highest loading rate.

Key value for chemical safety assessment:

EC50, EC10 > 100 mg/L, 100 mg/L entered in respective field below

Key value for chemical safety assessment

EC50 for freshwater algae:

100 mg/L

EC10 or NOEC for freshwater algae:

100 mg/L