Acetic acid, C11-14-branched alkyl esters, C13-rich

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:

06 August 1985 to 10 August 1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

According to the analytical results it can not be assured if a sufficient amount of test substance was used within the test.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

-Effect concentration based on % water soluble fraction (WSF)

GLP compliance:

yes

Remarks:

EPA good Laboratory Practice Regulations set forth in 40 CFR Part 792.

Test material

Test material information

Constituent 1

Reference substance name:

Acetic acid, C11-14-isoalkyl esters, C13-rich

EC Number:

283-740-9

EC Name:

Acetic acid, C11-14-isoalkyl esters, C13-rich

Cas Number:

84712-50-5

Molecular formula:

C15H30O2

IUPAC Name:

Acetic acid, C11-14-isoalkyl esters, C13-rich

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch I


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: The stability, identity, strength, and composition or other characteristics which appropriately identify the test substance are the responsibility of the Sponsor.


FORM AS APPLIED IN THE TEST (if different from that of starting material): A colorless liquid (not different from starting material)

OTHER SPECIFICS: The methods of synthesis, fabrication, and/or derivation of the test substance are documented and are the responsibility of the Sponsor.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control, 6.25, 12.5, 25, 50 and 100%
- Sampling method: Samples for test material concentration verification were taken from the AAP nutrient media, prior to test material preparation, from all test treatments prior to adding test organisms, and from all treatments at study termination.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The MRD-84-521 water soluble fraction was prepared by combining 6.7 mL of test material with 1 liter of AAP nutrient media in a 2 liter flask. This mixture was slowly stirred for approximately 72 hours. After the mixing period, the mixture was
transferred to a separatory funnel and allowed to settle for approximately 1 hour. After the settling period, the water soluble fraction was drawn off and used as the 100 % stock solution.

Test organisms

Test organisms (species):

Scenedesmus capricornutum

Details on test organisms:

TEST ORGANISM
- Common name: Scenedesmus capricornutum
- Strain: Initial strain 1648
- Source (laboratory, culture collection): Department of Botany, University of Texas.
- Age of inoculum (at test initiation): The study was started with algal cells in the log phase of growth as calculated from growth of stock cultures. This is documented in the Algal Culture Log.
- Method of cultivation: The algae are cultured in AAP nutrient media prepared with distilled water and reagent grade chemicals. The media was prepared as per standard laboratory procedures. Culture vessels were sterile glass flasks whose volume
was approximately 60 % of capacity. The cultures were maintained in 400 (+/- 10%) footcandles of continuous cool white light, on a stirring platform with aeration, at a continuous temperature of 21 (+/- 1) degrees C. Stock cultures were maintained for approximately 14 days with new stock prepared weekly. Additional culture information is available in the Algal Culture Log at EHSL.

ACCLIMATION
- Culturing media and conditions (same as test or not): Same as test

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Test temperature:

23.89 - 23.91

pH:

7.5 (+/- 0.1)

Nominal and measured concentrations:

Nominal: Control, 6.25, 12.5, 25, 50, 100 (% WAF)
Measured: -, 0.058, 0.219, 0.492, 0.873 (ppm total carbon)

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: size: 125 mL, fill volume: 50 mL
- Initial cells density: 2 X 10^4 cells/mL.
- Control end cells density: mean: 5.2 x 10^6 cells/mL.
- No. of vessels per concentration (replicates): 3 replicates
- No. of vessels per control (replicates): 3 replicates

GROWTH MEDIUM
- Standard medium used: yes (Miller, et al., 1978. The Selenastrum capricornutum Printz Algal Assay Bottle Test. U.S. EPA pp.17-18.)

TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: The pH was adjusted where necessary to 7.5 (+/- 0.1) prior to adding test organisms.
- Photoperiod: 24 hours
- Light intensity and quality: 400 footcandles.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: The fluorescence readings were converted to cell numbers using a regression formula developed through cell counts (hemocytometer).
- Chlorophyll measurement: 24, 48, 72, and 96 hours

TEST CONCENTRATIONS
- Range finding study: no

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

growth rate

Remarks on result:

other: results statistically not evaluable

Details on results:

- Exponential growth in the control (for algal test): yes

Any other information on results incl. tables

No EC0 or EC50 values can be reported as there was insufficient growth inhibition in all of the test treatments. An earlier trial of this study was performed, however several of the treatments exhibited an unacceptable degree of variability in growth. In the past, this variability has been associated with contaminated glassware. The measured concentration of the test material in solution during the study is described by the Day 0 values only. The Total Carbon measurement is very sensitive to changes in carbonate and CO2 in solution. As the solution Total Carbon on Day 4 is the result of algal metabolism during the study, it is not a true representation of the test material in solution.

Table 1: Mean Cell Concentrations (cells/mL)

Date	Control	6.25	12.5	25.0	50.0	100.0
06-AUG-85	2.5 x 10^4	2.6 x 10^4	2.7 x 10^4	2.6 x 10^4	2.4 x 10^4	2.4 x 10^4
07-AUG-85	8.2 x 10^4	8.0 x 10^4	8.9 x 10^4	8.2 x 10^4	8.5 x 10^4	8.6 x 10^4
08-AUG-85	4.8 x 10^5	5.0 x 10^5	5.3 x 10^5	4.8 x 10^5	4.9 x 10^5	5.7 x 10^5
09-AUG-85	2.5 x 10^6	2.6 x 10^6	2.6 x 10^6	2.2 x 10^6	2.3 x 10^6	2.4 x 10^6
10-AUG-85	5.2 x 10^6	4.4 x 10^6	4.6 x 10^6	4.1 x 10^6	5.3x 10^6	5.2 x 10^6

 

Table 2: Total Carbon Content (ppm)

Sample	06-Aug-85 Day 0 (unfiltered)	10-Aug-85 Day 4 (filtered)
Control	3.374	10.01
6.25%	3.282	9.452
12.5%	3.432	9.829
25%	3.593	9.648
50%	3.866	9.195
100%	4.247	9.513

Applicant's summary and conclusion

Validity criteria fulfilled:

not specified

Conclusions:

Based on the estimated 96-h ErC50 of >100 mg/L, the test material is not classified as acutely toxic to freshwater algae according to Regulation (EC) No. 1272/2008 (CLP).

Executive summary:

No ErC0 or ErC50 values can be reported as there was insufficient growth inhibition in all of the test treatments after 96 hours. Therefore a 96 -h EC50 value of > 100 mg/L was estimated.