Reaction mass of (Z)-1-chloro-1-propene and 2-chloropropane and 2-chloropropene

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study performed according to OECD guideline 202

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-chloropropene

EC Number:

209-187-5

EC Name:

2-chloropropene

Cas Number:

557-98-2

Molecular formula:

C3H5Cl

IUPAC Name:

2-chloroprop-1-ene

Constituent 2

Reference substance name:

2-chloropropane

EC Number:

200-858-8

EC Name:

2-chloropropane

Cas Number:

75-29-6

Molecular formula:

C3H7Cl

IUPAC Name:

2-chloropropane

Constituent 3

Reference substance name:

1-chloropropene

EC Number:

209-675-8

EC Name:

1-chloropropene

Cas Number:

590-21-6

IUPAC Name:

1-chloroprop-1-ene

Details on test material:

Identification Reaction mass containing mainly 2-chloropropene
Molecular formula C3H5Cl
Molecular weight 76.53
CAS Number 557-98-2
General information Extremely flammable
Test substance handling If necessary, handling after sampling will be performed on dry ice in order to keep the test substance liquefied
Density 0.9
Stability in water: Not indicated
Solubility in water: Yes

Description Clear light yellow to brown liquid (determined at NOTOX)
Batch RBA100301A
Purity Minimum 60.0%
Test substance storage In refrigerator (2-8°C) in the dark
Stability under storage conditions Stable
Expiry date 08 March 2011 (allocated by NOTOX, 1 year after receipt of the test substance)

The vapour pressure of the test substance at 20°C (293K) was 84.9  103 Pa = 637 mm Hg. The water solubility of the test substance at 20°C was 1.94 g/l (determined with the flask method)(NOTOX project 492885).

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Duplicate samples for possible analysis were taken from all test concentrations and the control according to the schedule below.

Frequency at t=0 h and t=48 h
Volume 10 ml from the approximate centre of the test vessels
Storage Samples were stored in a freezer until analysis.

At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Additionally, duplicate reserve samples of 10 ml were taken for possible analysis. If not used, these samples were stored in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.

Test solutions

Vehicle:

no

Details on test solutions:

The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g. film of the test substance on the water surface).

The batch of Reaction mass containing mainly 2-chloropropene tested was a clear light yellow to brown liquid with a minimum purity of 60% and the substance was completely soluble in test medium at the concentrations tested.

Preparation started with a concentration of 100 mg/l; 111 µl of the test substance was added to 1000 ml medium and the vessel was directly air-tight closed. A 15-minute period of magnetic stirring was applied to accelerate the dissolving of the test substance in the test medium. The lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium. The final test solutions were all clear and colourless.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.

Source: In-house laboratory culture with a known history.

Reason for selection : This system has been selected as an internationally accepted invertebrate species.

Validity of batch: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

Characteristics: For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.

Breeding:
Start of each batch With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.

Maximum age of the cultures 4 weeks

Renewal of the cultures After 7 days of cultivation half of the medium twice a week.

Temperature of medium 18-22°C

Feeding Daily, a suspension of fresh water algae.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

The test was not conducted more longer due to the high volatility of the test substance.

Test conditions

Hardness:

180 mg/l expressed as CaCO3

Test temperature:

18-22 °C

pH:

Start of test: 7.9 - 8.0
End of the test: 8.0 - 8.2

Dissolved oxygen:

Start of test: 8.9 - 9.2 mg/l
End of the test: 7.5 - 8.4 mg/l

Salinity:

Macro salts: CaCl2.2H2O 211.5 mg/l
MgSO4.7H2O 88.8 mg/l
NaHCO3 46.7 mg/l
KCl 4.2 mg/l

Nominal and measured concentrations:

32, 56 and 100 mg/l (nominal concentration) corresponding respectively to 5.3, 8.2 and 28 mg/l (mean measured concentration)

Details on test conditions:

Combined limit/ range-finding test:
The project started with a combined limit/range-finding test. Twenty daphnids per concentration (four replicates, 5 daphnids per replicate) were exposed to a control and a concentration of 100 mg/l. Test procedure and conditions were similar to those applied in the final test with the following exceptions:
• Ten daphnids per concentration (in duplicate, 5 per vessel) were exposed to 0.1, 1.0 and 10 mg/l in the combined range-finding test.
• Dissolved oxygen concentrations and pH were only measured in the control and the highest test concentration.
• Test was performed in 100 ml all glass vessels closed with a plastic screw-cap and filled up to the edge with test solution.

Final test:
-Test concentrations:
Reaction mass containing mainly 2-chloropropene 10, 18, 32, 56 and 100 mg/l
Controls Test medium without test substance or other additives
-Test procedure and conditions:
Test duration 48 hours
Test type Static
Test vessels 50 ml, all-glass, closed air-tight. In order to minimize the headspace the vessels were filled with the test solution up to the edge.
Medium Adjusted ISO medium
Number of daphnids 20 per concentration
Loading 5 per vessel containing approximately 50 ml of test solution
Light 16 hours photoperiod daily
Feeding No feeding

Aeration No aeration of the test solutions.

Introduction of daphnids Within 16 minutes after preparation of the test solutions.

Reference substance (positive control):

yes

Remarks:

K2Cr2O7-Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

7.7 mg/L

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

8.2 mg/L

Effect concentrations 3

Dose descriptor:

NOEC

Effect conc.:

5.3 mg/L

Details on results:

1) Combined limit/range-finding test:
Table 1 shows the responses recorded during the combined limit/range-finding test. No significant immobility was observed at the test concentrations from 0.1 to 10 mg/l and in the control during the test period. At the highest concentration all daphnids were immobilized.
Based on these results samples taken from nominal 10 mg/l were analysed. The mean initial concentration was 2.3 mg/l. The measured concentration remained constant during test period (103% of initial, see also Table 2 of the appended Analytical Report).
The expected EC50 was between 10 and 100 mg/l (based on the nominal concentrations).

All test conditions were maintained within the limits prescribed by the protocol.

2) Final test:

Measured concentrations:

Analysis of the samples taken at the start of the final test showed mean measured concentrations of 6.4, 11 and 28 mg/l at the nominal concentrations of 32, 56 and 100 mg/l, respectively. Measured concentrations after 48 hours of exposure were 4.4, 5.9 and 29 mg/l, respectively. Based on these results, the average exposure concentrations were 5.3, 8.2 and 28 mg/l.

Immobility:

Table 2 shows the responses recorded during the final test.

The responses recorded in this test allowed for reliable determination of an EC50. The responses recorded at the nominal concentrations of 10 and 100 mg/l were in agreement with the results of the combined limit/range-finding test.

Determination of effect concentrations:

Table 3 shows the effect parameters based on the average exposure concentrations.

Experimental conditions:

The results of measurement of pH and oxygen concentrations (mg/l) are presented in Table 4. These test conditions remained within the limits prescribed by the protocol (pH: 6.0-8.5, not varying by more than 1.5 unit; oxygen: >3 mg/l at the end of the test).

The temperature of the test medium was 20.4°C at the start of the test. The temperature continuously measured in a temperature control vessel varied between 18.7 and 20.2°C during the test, and complied with the requirements as laid down in the protocol (18-22°C, constant within 2°C).

Results with reference substance (positive control):

The 24h-EC50 was 0.75 mg/l with a 95% confidence interval between 0.56 and 1.0 mg/l.

The 48h-EC50 was 0.36 mg/l with a 95% confidence interval between 0.28 and 0.49 mg/l.

Any other information on results incl. tables

Table 1           Incidence of immobility in the combined limit/range-finding test

Concentration test substance^1 (mg/l)	Vessel number	Number Daphnia exposed	Response at 24 h		Response at 48 h
			number	Total %	number	Total %
control	A B C D	5 5 5 5	0 0 0 0	0	0 0 0 0	0
0.1	A B	5 5	0 0	0	0 0	0
1.0	A B	5 5	1 0	10	1 0	10
10 (2.3)	A B	5 5	0 0	0	0 0	0
100	A B C D	5 5 5 5	5 5 5 5	100	5 5 5 5	100

1. Reaction mass containing mainly 2-chloropropene

() – initially measured concentration

Table2           Acute immobilisation of daphnids after 24 and 48 hours in the final test

Concentration test substance^1 (mg/l)	Vessel number	Number Daphnia exposed	Response at 24 h		Response at 48 h
			number	Total %	number	Total %
control	A  B C D	5 5 5 5	0 0 0 0	0	0 0 0 0	0
10	A  B C D	5 5 5 5	0 0 0 0	0	0 0 0 0	0
28	A  B C D	5 5 5 5	0 0 0 0	0	0 0 0 0	0
32 (5.3)	A  B C D	5 5 5 5	0 0 0 0	0	0 0 0 0	0
56 (8.2)	A  B C D	5 5 5 5	2 3 3 2	50	3 4 3 3	65
100 (28)	A  B C D	5 5 5 5	5 5 5 5	100	5 5 5 5	100

1. Reaction mass containing mainly 2-chloropropene

() – between brackets the average exposure concentrations are given

Table3           Effect parameters

Parameter	Average exposure concentration test substance^1 (mg/l)	95%-confidence interval
NOEC	5.3
24h-EC50	8.2
48h-EC50	7.7	7.1 – 8.6

1. Reaction mass containing mainly 2-chloropropene

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present study Reaction mass containing mainly 2-chloropropene did not induce acute immobilisation of Daphnia magna at an average exposure concentration of 5.3 mg/l after 48 hours of exposure (NOEC).

The 48h-EC50 was 7.7 mg/l based on an average exposure concentration (95% confidence interval between 7.1 and 8.6 mg/l).

Executive summary:

The study procedures described in this report were based on the OECD guideline No. 202, 2004. In addition, the procedures were designed to meet the test methods of theCommissionRegulation (EC) No440/2008,Part C.2, 2008 and the ISO International Standard 6341.

 

The batch of Reaction mass containing mainly 2-chloropropene tested was a clear light yellow to brown liquid with a minimum purity of 60% and the substance was completely soluble in test medium at the concentrations tested.

 

Preparation started with a concentration of 100 mg/l. The lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium.

 

A final test was performed based on the results of a combined limit/range-finding test. Twenty daphnids per test group (5 per vessel, 4 vessels) were exposed to a control and concentrations of 10, 18, 32, 56 and 100 mg Reaction mass containing mainly 2-chloropropene per litre. The total exposure period was 48 hours and samples for the analytical confirmation of the exposure concentrations were taken at the start and at the end of the test. Both tests, due to the volatile nature of the test substance, were performed in air-tight closed vessels with a minimal headspace.

 

Analysis of the samples taken at the start of the final test showed mean measured concentrations of 6.4, 11 and 28 mg/l at the nominal concentrations of 32, 56 and 100 mg/l, respectively. Measured concentrationsafter 48 hours of exposure were 4.4, 5.9 and 29 mg/l, respectively. Based on these results, the average exposure concentrations were 5.3, 8.2 and 28 mg/l.

 

The study met the acceptability criteria prescribed by the protocol and was considered valid.

 

Reaction mass containing mainly 2-chloropropene did not induce acute immobilisation of Daphnia magna at an average exposure concentration of 5.3 mg/l after 48 hours of exposure (NOEC).

 

The 48h-EC₅₀was 7.7 mg/l based on an average exposure concentration (95% confidence interval between 7.1 and 8.6 mg/l).