Chloro(1-methylethyl)magnesium

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:

2012-10-26 to 2013-06-14

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

, adopted on 17th July 1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: "No. 203 "Fish, Acute Toxicity Test" of "The Guidelines for the Testing of Chemicals" (2004) Ministry of Environment Protection the people's Republic China

Deviations:

no

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Chloro(1-methylethyl)magnesium

EC Number:

213-947-1

EC Name:

Chloro(1-methylethyl)magnesium

Cas Number:

1068-55-9

Molecular formula:

C3H7ClMg

IUPAC Name:

chloro(1-methylethyl)magnesium

Constituent 2

Reference substance name:

Isopropylmagnesium chloride

IUPAC Name:

Isopropylmagnesium chloride

Test material form:

other: liquid: 20% in Tetrahydrofuran

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Testconcentration was messeaured at beginning and the end of the test c(96h)/c(0h) = 91.66 - 105.3%

Test solutions

Details on test solutions:

PREPARATION AND APPLICATION OF STOCKSOLUTION:

2500µL Isopropyl magnesium chloride ca. 20% in Tetrahydrofuran (0.5 g Isopropyl magnesium chloride) was added into a 1000 mL beaker (contained 50 mL pure water). 1L test water was added and treated with ultrasonic dissolving for 20 min. Thereafter, after filtrated through 30 µm filter membrane, it was transferred into an aquarium and diluted to 6 L with test water. 1 mol/L HNO3 was added to adjust the pH value to 7.8 +- 0.1 and it was stirred with a glass rod for 2 min to obtain 100 mg/L test solution .

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Brachydanio rerio
- Source: Shanghai Hongye Aquarium Fish Breeding Farm
- Age at study initiation: 2-3 months
- Length at study initiation: 2.2 - 2.8 cm
- Weight at study initiation: 0.12 -0.18 g
- Feeding during test: no


ACCLIMATION
- Acclimation period: 2012-12-10 to 2012-11-16
- Acclimation conditions (same as test or not): yes, exception: during the test no feeding
- Type and amount of food: Ingredients: Crude Protein > 35%, Crude Fat > 2%, Crude Fibre < 10%, Crude Ash < 18%, Moisture < 10%, Calcium > 0.3%, Phosphorus > 0.4%, Lysine > 0.6%
- Feeding frequency: 0.2 g once a day, until 24 h before the test was started
- Health during acclimation (any mortality observed): no mortality

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Test temperature:

Temperaturrange during 96 h test period: 23.5 - 24.5°C
Temperature complied with request of guidlines.

pH:

Range of pH during 96 h test period: 7.68 - 7.88
Ph complied with request of guidlines.

Dissolved oxygen:

Range of dissolved oxygen during 96 h test period: 81.2 - 105.4%
Concentration complied with request of guidlines.

Nominal and measured concentrations:

Nominal concentrations of the preliminaray test: 0, 1.0, 10.0, 100.0 mg/L Isopropyl magnesium chlorid
Nominal concentrations of limit test: 100.00 mg/L Isopropyl magnesium chlorid

Details on test conditions:

TEST SYSTEM
- Aquariums with 5L test solution
- Aeration: for 24 hours before the test started
- No. of organisms per vessel: 10 (but not more than 1g fish in 1L water)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Dead fish were taken out in time.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- 58.80 g CaCl2 * 2H20 was dissolved in deionized water and was diluted to 5L
- 12.95 g NaHCO3 was dissolved in deionized water and was diluted to 5L
- 1.15 g KCl was dissolved in deionized water and was diluted to 5L
The above three solutions were mixed and diluted to 20 L with deionized water.
Subsequently and after 24 houres of aeration physicochemical parameters were measured.

OTHER TEST CONDITIONS
- Adjustment of pH: 1 mol/L HCl to adjust the pH value to 7.8 +- 0.1
- Photoperiod: Natural light (12 hours light/ 12 hours dark)
- Temperature: 24 +- 1°C

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortalites and responses of fish, water temperature, pH and dissolved oxygen recorded every 24 h

TEST CONCENTRATIONS

- Test concentrations: Limit Test 100 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromat

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: no
- Mortality of control: 0

Results with reference substance (positive control):

LC50 (24h) = 248.2 mg/L (95% confidence limits 213.7 - 288.2 mg/L)
This is in accordance with the accepted 200 - 400 mg/L range, which satisfied the test requirement. The results of reference substance are valid.

Reported statistics and error estimates:

LC50 (24h) and confidence limits (p=0.95) of positive control were calculated by modified linear regression method according to the measured concentration values. Probit function was used.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

During test period no dead fish was observed. The result indicated that the LC50(96h) was greater than 100 mg/L (highest nominal observed testconcentration).

Executive summary:

This study was conducted to determine the acute toxicity of the test substance isopropyl magnesium chloride to Brachydanio rerio after 96 h testperiod. Resulst of preliminary test indicated that there was no dead fish and the fish had no abnormal behavior during the test period under nominal concentrations of 1, 10 and 100 mg/L Isopropyl magnesium chlorid. Therefore it was not necessary to conduct the definitve test. However limit test with 100 mg/L testsolution was carried out for three replicates. Each replicate consists of ten organisms in 5 L aquarium.

The results indicated after 96 h exposure, that there was no dead fish and no abnormal behavior observed during the period. This demonstrated that the LC50 (96h) of Isopropyl magnesium chloride for fish was greater than highest tested nominal concentration of 100 mg/L. Potassium dichromat was used as positive control. The calculated LC50 (24h) = 213.7 -288.3 mg/L is in accordance with the accepted 200 -400 mg/L range which satisfied the tes requirement.