(3S-cis)-3,6-dimethyl-1,4-dioxane-2,5-dione

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Recent GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Remarks:

The test method was modified to involve a better buffered medium (150 mg.H NaHCOs) and the use of a parametric model (3) to calculate the effect concentrations.

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

L-(+)-lactic acid

EC Number:

201-196-2

EC Name:

L-(+)-lactic acid

Cas Number:

79-33-4

IUPAC Name:

2-hydroxypropanoic acid

Details on test material:

80% Lactic acid solution in water. Batch no. ZO 3456

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Samples were taken at t=0 from 0, 0.1, 1.0 and 2.8 g/L nominal concentrations
- Sampling method: 250 mL bottles
- Sample storage conditions before analysis: Each sample was analysed twice (on October 2 and November 2,1991); the samples were stored in a refrigerator pending analysis.

Test organisms

Test organisms (species):

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

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Post exposure observation period:

None

Test conditions

pH:

Buffered solutions

Nominal and measured concentrations:

Nominal (test material) 0,0.1,0.33,0.56,1.0,1.9 and 2.8 g/L; nominal (active substance) 0, 0.08, 0.26, 0.45, 0.8, 1.72, 2.24 g/L; actual (active substance) <0.005, 0.020-0.065, 0.44-0.62, 1.45-1.74 g/L (for 0, 0.1, 1.0 and 2.8 mg/L nominal concentrations)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed
- Initial cells density: 0.9e4 cells/mL
- Control end cells density: 156e4 cells/mL
- No. of vessels per concentration (replicates):2
- No. of vessels per control (replicates): 4


GROWTH MEDIUM
- Standard medium used: yes


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [electronic particle counter]


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study
- Test concentrations: 0, 0.1, 1.0, 10, 101 and 1008 mg/L
- Results used to determine the conditions for the definitive study:

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

70 h

Dose descriptor:

NOEC

Effect conc.:

1.9 g/L

Basis for effect:

growth rate

Effect concentrations 2

Duration:

70 h

Dose descriptor:

EC50

Effect conc.:

3.5 g/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 2.8 g/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

By visual comparison of the measured and the calculated growth curves of algal suspen- sions exposed to the test substance with those of the algal controls (see Fig. 1), the no- observed-effect-concentration (NOEC) of the 80% aqueous solution of L(+)lactic acid was estimated to be 1.9 g.1'1.
Microscopic examination of the cells at the start and end of the incubation period revealed no abnormalities, however many bacteria were found in the cultures exposed to test substance concentrations of 1.0,1.9 and2.8 g.1-1.
The pH of the algal medium containing different test substance concentrations was adjusted to the range pH 7.5-8.0 at the start of the test. It was found to remain constant (pH 8.0-8.1) during the test. In the presence of algae the pH was found to increase a little with algal cell density (pH 8.2-8.7).

Any other information on results incl. tables

Results are based on nominal concentrations.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

NOEC of lactic acid to green algae is 1.52 g/L nominal concentration of active substance.

Executive summary:

The toxicity of a solution of about 80 % L(+)lactic acid to the fresh-water green alga Selenastrum capricornutum was determined in a growth inhibition test according to the OECD Guideline no. 201 and the OECD principles of Good Laboratory Practice. The test included chemical determinations of the test substance concentrations during the test.

The nominal concentrations of neutralized 80 % L(+)lactic acid tested were O, 0.10, 0.33, 0.56, 1.0, 1.9 and 2.8 g .H . The algal growth was detennined by electronic partiële counting. The effect values were calculated using a parametric model assuming an error proportional to the number of cells.

The EC50 with respect to inoculum viability followed by exponential growth (EeC50) was found to be 3.5 g.H, with a 95% confidence interval of 2.8-4.4 g.H. The corresponding EeC10 and EeC90 values were 2.3 g.H and 5.4 g.H respectively. The EC50 with respect to the area under the growth curve (Et>C50) was found to be > 2.8g.H.ThecorrespondingEt,C10andEfcC90valueswere2.4g.H (intherange1.9-2.8 g.H) and > 2.8 g.1-1 respectively.

The no-observed-effect-concentration (NOEC) with respect to the effects referred to above was estimated to be 1.9 g.1-1.

An effect on the inoculum viability, as found in this test, may be expected when a substance looses its toxicity during the test, in this case probably due to biodegradation.

The concentrations quoted in thij; report are the nominal concentrations of the test substance (a solution of about 80 % L(+)lactic acid) as supplied by sponsor. At the start of the test 65-82% of the nominal test substance concentrations were found by chemical analysis. At the end of the test the concentrations were considerably lower: 15,57 and 73% at the three concentrations analysed.