diethyleneglycyl bis (chloroacetate)

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001-01-29 to 2001-02-02

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Principles of method if other than guideline:

Some test fish had a somewhat smaller size than the one recommended in the test guideline for the zebra fish.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-{2-[(2-chloroacetyl)oxy]ethoxy}ethyl 2-chloroacetate

Cas Number:

106-78-5

Molecular formula:

C8H12O5Cl2

IUPAC Name:

2-{2-[(2-chloroacetyl)oxy]ethoxy}ethyl 2-chloroacetate

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Sampling schedule of chemical analysis:
Stock solution: at 0 hours
Control: 0, 24, 48, 72 and 96 hours
Test concentrations: at 0, 24, 48, 72 and 96 hours

Test solutions

Vehicle:

no

Details on test solutions:

Pretreatment of the test substance:
A stock solution was prepared to give the desired series of test concentrations. To achieve this 1000.2 mg of the test substance were added to 10 litres ofdilution water, then treated with an ultra turrax for 60 seconds and afterwards stirred for 24 hours on a magnetic stirrer. Finally undissolved particles of the test substance were removed by filtration.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Strain: Zebra fish (Brachydania reria HAMILTON-BUCHANAN)
- Source: Bio International B. V. (Netherlands)
- Age (mean and range, SD): Date ofbirth: March 9, 2000; Date supplied: July 25,2000
- Length (length definition, mean, range and SD): mean standard length (n = 60): 2.7 cm (S.D. = 0.29 cm)

ACCLIMATION
- Acclimation period: Stock held since July 25,2000 and acclimatised to the test conditions since then
- Acclimation conditions (same as test or not): not specified
- Health during acclimation (any mortality observed): Mortality <2%
- Feeding: Commercial fish food, daily. Discontinued 24 h prior to test


FEEDING DURING TEST
no feeding during exposure period

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

The hardness of the dilution water used was 13.1 °dH (= 233.8 mg/I CaC03).

Test temperature:

20-24°C +- 1°C

pH:

6.8 - 7.9

Dissolved oxygen:

8.2 - 8.9 mg/L

Salinity:

n.a.

Conductivity:

no data

Nominal and measured concentrations:

See table 1

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria holding 5 I oftest media
- Type (delete if not applicable): open
- Aeration: gentle aeration via narrow glass tubes
- Renewal rate of test solution (frequency/flow rate): none
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.70g bw (wet weight) per litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: prepared according to ISO 7346
- Total organic carbon: see table 1
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Photoperiod: 16h lieght : 8h dark


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
In addition to observations on mortality at 2, 24, 48, 72 and 96 hours, subjective assessments were made on the type and incidence ofsub-lethal effects compared with control fish.

TEST CONCENTRATIONS
6.3, 12.5, 25, 50 and 100 mg/L

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

13.5 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

52.7 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1: Chemical analysis

Time	0h	24 h	48 h	72 h	96 h
Control
TOC values [mg/L]	<2	<2	<2	3	3
Nominal concentration [mg/L]: 6.3
TOC values [mg/L]	3	3	4	3	4
measured test substance [mg/L]	8.1	8.1	10.8	8.1	10.8
Nominal concentration [mg/L]: 12.5
TOC values [mg/L]	5	6	6	5	3
measured test substance [mg/L]	13.5	16.2	16.2	13.5	8.1
Nominal concentration [mg/L]: 25
TOC values [mg/L]	10	11	13	11	11
test substance [mg/L]	27	29.7	35.1	29.7	29.7
Nominal concentration [mg/L]: 50
TOC values [mg/L]	19	20
measured test substance [mg/L]	51.3	54
Nominal concentration [mg/L]: 100
TOC values [mg/L]	37	40
measured test substance [mg/L]	99.9	108

Table 2: Analysis of mortality data give the following results:

Time [h]	LC 0 [mg/L]	LC 100 [mg/L]	Geometric mean LC 0 / LC 100 [mg/L]
2	>= 99.9	> 99.9	-
24	14.9	52.7	28.0
48	15.3	52.7	28.4
72	14.9	52.7	28.0
96	13.5	52.7	26.7

 

Highest test concentration resulting in 0% mortality (LC 0 96h): 13.5 mg/L

Lowest test concentration resulting in 100% mortality (LC 100 96h): 52.7 mg/L

All results are expressed in terms of mean measured concentrations. Measured concentrations ranged from 99.9 - 128.6% of nominal values at 0 hours, from 108.0 - 129.6% of nominal values at 24 hours, from 129.6 -171.4% ofnominal values at 48 hours, from 108.0 -128.6% of nominal values at 72 hours, and from 64.8 - 171.4% of nominal values at 96 hours, respectively.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions a LC 0 96 h of 13.5 mg/L and a LC 100 96h of 52.7 mg/L was determined.